Pharmacognosy Magazine

Ferroptosis is a novel type of regulated cell death and targeting ferroptosis may be a potential treatment strategy for lung cancer. Ziyuglycoside II (ZYG II) has a significant inhibitory effect on the growth of lung cancer cells. However, the selective anti-tumor effect of the ZYG II against lung cancer has not been systemically studied.

We combined ferrostatin-1 and erastin to explore the potential therapeutic mechanism of the ZYG II for lung adenocarcinoma.

A549 and H1299 cells were randomly divided into the control, ZYG II, ferroptosis inhibitor group (ZYG II+ ferrostatin-1), and erastin group (ZYG II+ erastin). Cell proliferation was detected using the CCK-8 method. Cell migration and invasion were evaluated using the Transwell assay. The protein expression levels of Glutathione Peroxidase 4 (GPX4), Solute Carrier Family 7 Member 11 (SLC7A11), and Transferrin receptor 1 (TFR1) were measured using western blotting.

Compared with the control group, the cell proliferation, migration, and invasion abilities of the ZYG II group significantly decreased, the protein expression levels of GPX4 and SLC7A11 in the ZYG II group declined significantly, and the expression of TFR1 increased significantly (p < 0.05). After adding ferrostatin 1 (ZYG II+ Ferrostatin 1), the cell proliferation, migration, and invasion abilities of the inhibited cells were significantly increased, the expression of GPX4 and SLC7A11 increased significantly and the expression of TFR1 decreased significantly (p < 0.05). However, after adding the erastin (ZYG II+ erastin), the cell viability was further inhibited in A549, the expression levels of GPX4 and SLC7A11 were further inhibited and the expression of TFR1 was further increased (p < 0.05).

ZYG II significantly inhibited the survival rate, proliferation, migration, and invasion ability of A549 and H1299 cells, possibly by inducing ferroptosis.

The aim of this study was to investigate the effect of Panicum miliaceum involved in the repair of femur bone fracture in rats.

In this study, 40 Wistar rats in five groups were studied as healthy, rats with lesion induced by surgery in femur (FX), FX rats treated with P. miliaceum extract (FX +100 and 200 P. miliaceum) and FX rats treated with Osteocare syrup (FX + OC). After 40 days, the serum levels of blood calcium (Ca), phosphorus (P), and alkaline phosphatase (ALP) along with activity of glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT) enzymes, and nitric oxide (NO) level were measured. The serum level of cytokines (IL-6, C-reactive protein [CRP], IL-10, IL-1β, and tumor necrosis factor-α [TNF-α]) was also measured by ELISA. The expression of OPG, RANK, RANKL, and BMP-2 genes and proteins were evaluated by real-time PCR and Western blotting techniques, respectively.

The results showed that P. miliaceum was able to decrease the activity of ALP enzyme in addition to increasing serum Ca and P levels and GPx, SOD, and CAT activity. P. miliaceum reduces serum IL-6, IL-1β, and TNF-α, and NO levels. Also, the extract of this plant could increase the expression of OPG/RANK/BMP-2 pathway genes and proteins.

P. miliaceum can be used as a prodrug in traditional medicine to repair bone fractures and osteoporosis.

The prevalence of obesity is rising worldwide. Due to the side effects associated with modern drugs, there is a need for an alternative treatment for obesity management. Triphala (TP), a polyherbal formulation from Ayurveda, is used by Ayurvedic physicians for weight loss.

This study was designed to evaluate the effects of TP on obesity at the biochemical, histological, and molecular levels.

Male Wistar rats (125–150 g, 6–7 weeks old) were fed with a high-fat diet (HFD) for total of 48 days. From day 24 to day 48, along with HFD, rats were given an aqueous extract of TP (50, 100, and 200 mg/kg/day) or atorvastatin (1.2 mg/kg/day). Biochemical parameters were estimated in serum. Histopathology was done for the liver and adipose tissue. Levels of the genes involved in lipid metabolism were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Monoamine levels were estimated from the brain tissue.

At the end of the study (day 48), compared to only HFD receiving group (DC), all TP-treated groups showed a significant decrease in body weight, serum glucose, total cholesterol, triglycerides, leptin, interleukin-6, C-reactive protein, malondialdehyde, and noradrenaline levels. High-density lipoprotein, adiponectin, superoxide dismutase, serotonin, and dopamine levels were found to be increased compared to DC rats. Expression of various genes involved in lipid metabolism was found to be down regulated only in TP 100 group compared with DC. Additionally, all TP groups showed a reduction in adipocyte size and restored monoamine levels.

The aqueous extract of TP had shown an anti-obesity effect as demonstrated by lowering inflammation and oxidative stress, adipocyte size, and modulation of expression of genes involved in lipid metabolism.

Non-alcoholic fatty liver disease (NAFLD) is severely affecting the quality of people’s life. Rhapontigenin (RA) is a stilbene compound isolated from Rhubarb L., which has been reported to have an effect on cholesterol diet-induced hyperlipidemia in rats. This study aims to explore the pharmacodynamics and the mechanism of RA obtained from Rheum franzenbachii Munt. against NAFLD. RA was extracted from the roots of the Rheum L. (Polygonaceae) plant Rheum franzenbachii Munt.

RA was extracted by Sephadex-gel column and identified by high-performance liquid chromatography (HPLC)-UV and HR-ESI-MS. The pharmacodynamic indexes of L-02 cells and mice treated with RA were determined by histological staining and ELISA, while the expression of autophagy-related proteins was analyzed by Western blot.

The results in vivo showed that the liver structure of RA-treatment mice was normal, and the organ coefficient was significantly decreased. It also showed that RA could significantly reduce the expression of reactive oxygen species (ROS) in the liver as well as inhibit oxidative stress and inflammatory response. Interestingly, the autophagy inhibitor 3-methyladenine (3-MA) could reverse the effect of RA on NAFLD, which further confirmed that RA plays an anti-NAFLD role through activating lipophagy.

It suggested that RA has an effect on NAFLD by down-regulating the expression of Acyl-CoA oxidase 1 (ACOX1), p-mTOR, and p-Unc-51-like kinase 1 (ULK1), then inhibiting Acetyl-CoA (A-CoA) production, and up-regulating the expression of autophagy protein 5 (Atg5) to promote the lipophagy of lipocytes.

Medicinal plants standardization is more concerning for its regulatory aspects based on safety, quality, and efficacy. Nyctanthes arbor-tristris is an Indian-origin medicinal plant that is used for numerous acute and chronic diseases. Due to the lack of an ethnopharmacological perspective based on biomolecular mechanisms, this study is associated to explore quality-based standardization and biomolecular mechanism of Nyctanthes arbor-tristris phytochemicals as a therapeutic application regimen in liver disease and associated complications.

2,2-Diphenyl-1-picrylhydrazyl (DPPH) and iron chelating effect of prepared extract of Nyctanthes arbor-tristris was examined for antioxidant effect. High-performance thin-layer chromatography (HPTLC) and liquid chromatography and mass spectroscopy (LC–MS) qualitative and quantitative analysis was conducted to unravel metabolites of Nyctanthes arbor-tristris. Network pharmacology as well as insilico docking analysis were performed to examine molecular interaction of ligands and targeted genes that regulate liver malfunction.

The results revealed that Nyctanthes arbor-tristris significantly (p < 0.05) scavenge DPPH free radicals and iron chelating effect and thus exhibited an antioxidant effect. HPTLC and LC–MS analysis showed several major and minor metabolites Nyctanthes arbor-tristris the content of naringenin, ferulic acid, and caffeic acid was found to be 1.662 ± 0.027, 4.411 ± 0.201, and 4.846 ± 0.154, respectively. Network pharmacology and insilico docking analysis revealed the multi-targeted therapeutic effect of metabolites identified in Nyctanthes arbor-tristris against liver disease and associated pathophysiology’s via regulation in the expression of several genes such as nitric oxide synthase (NOS), tumor necrosis factor alpha (TNF-α), interleukins (ILs), toll-like receptors (TLRs) and serum aminotransferase.

The study concludes that Nyctanthes arbor-tristris play a multi-mechanistic and therapeutic action against liver-associated distortion and functional inability against oxidative and inflammatory stress, hepatocytes fibrosis, and apoptosis.

Glabridin is one of the main components of the isoflavonoids in Glycyrrhiza glabra and possesses anti-inflammatory, anti-bacterial, and anti-cancer effects.

Herein, the therapeutic effects and mechanisms of action of the glabridin liposome (GL) complex were studied in mice with imiquimod-induced psoriasis.

After treatment with GLs, their effectiveness was assessed using parameters, such as Psoriasis Area and Severity Index (PASI) score, histopathology, inflammatory cytokines, and psoriasis-associated proteins.

The results demonstrated that GLs could significantly improve psoriatic symptoms, downregulate mast cell infiltration, and significantly reduce the PASI score of psoriatic mice. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) results showed that GLs significantly decreased IL-23 and STAT3 mRNA expression. The results of immunohistochemistry and the enzyme-linked immunosorbent assay indicated that GLs decreased the expression of tumor necrosis factor-alpha (TNF-α), IL-17, and IL-22.

GLs can alleviate psoriasis by inhibiting the expression of keratinocyte-activating proteins. These results suggest that GLs have great potential for the clinical treatment of psoriasis.

Modern strategies to alleviate the harmful effects of organophosphate pesticide diazinon (DN) abnormalities were focused mainly on using natural compounds or their derivatives. DN is an organophosphate compound that causes many health abnormalities in humans due to its usage as an insecticide in agriculture. TQ is one of the beneficiary active principles derived from plant sources that has pharmacological benefits.

This research reveals the therapeutic potential of thymoquinone (TQ) on DN-induced myocardial infarction (MI) in rats.

Male Sprague–Dawley rats were procured, acclimatized, and divided into four groups of six animals each with feed and water ad libitum. MI was induced in rats with a dose of 25 mg/kg DN by oral gavage and TQ in a dose of 20 mg/kg p.o. for the treatment. After animal sacrifice at the end of the experimental period, serum and heart tissue samples were collected and processed appropriately for various analyses such as changes in the body weight, heart weight, marker enzymes, oxidative markers, non-enzymatic and enzymatic antioxidants, inflammatory cytokines, histopathological studies, and cardiac-specific markers.

DN-induced toxicity depicted decreased body weight (167.83 ± 4.62), heart weight (0.9 ± 0.06), and heart-to-body weight ratio (0.54 ± 0.03). Also, elevated marker enzymes (147.33 ± 20.85, 407.5 ± 31.3, and 110.67 ± 9.65 for CK-MB, AST, and ALT, respectively), elevated oxidative markers (12.87 ± 1.34, 125.17 ± 9.95, and 80.17 ± 5.78 for serum MDA, heart MDA, and heart GSSG, respectively), decreased enzymatic- and non-enzymatic antioxidants (3.15 ± 0.42, 12.23 ± 1.02, 5.75 ± 0.46, 2.02 ± 0.26, 0.72 ± 0.07, 18.05 ± 1.04, 8.62 ± 0.65, and 45.8 ± 2.43 for SOD, CAT, GST, GPx, heart GSH, serum GSH, vit.E, and vit.C, respectively), damaged cellular architecture, elevated inflammatory cytokines, and cardiac-specific markers were noticed.

TQ significantly reduced the toxicities produced by DN in almost all the above parameters. The beneficial effect of DN could be attributed to the influential effect of DN on cardiac-specific Nrf2/HO1-related pathways.

These results suggest that TQ exerts protection against MI and could serve as a promising candidate for drug development.

Psoriasis vulgaris is an immune-mediated inflammatory skin disease that is associated with depression. In this study, we investigated the effect of Xiegan–Liangxue–Jiedu (XGLXJD) decoction, a traditional Chinese medicine formula, on psoriasis based on network pharmacology, molecular docking, and animal experiments.

A protein–protein interaction (PPI) network was constructed using the overlapping targets of XGLXJD decoction and psoriasis. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed using Metascape database. High-performance liquid chromatography (HPLC) was used to investigate the main compounds of XGLXJD decoction. Molecular docking was performed to predict the potential interaction between the main compounds and proteins of interest. A C57 mouse model of psoriasis was established via continuous exposure to imiquimod. Seven days later, the XGLXJD decoction was orally administered at increasing doses for 6 days. The psoriasis area and severity index were calculated. Hematoxylin and eosin staining was used to examine skin morphology. Enzyme-linked immunosorbent assay (ELISA) was used to determine the serum levels of adenylyl cyclase (AC), cyclic adenosine monophosphate (cAMP), protein kinase A (PKA), interleukin-17 (IL-17), and tumor necrosis factor-α (TNF-α). Sucrose preference test and forced swimming test were used to assess depression-like behavior. Immunohistochemical (IHC) staining and immunofluorescence were used to investigate the cAMP-response element binding protein (CREB) signaling pathway.

Overall, 162 overlapping targets were generated. A total of 398 biological processes, 84 molecular functions, and 47 cellular components were identified via GO analysis, whereas 140 pathways were identified via KEGG pathway analysis. The most notable signaling pathways were cAMP as well as downstream IL-17 and TNF-α signaling pathways. HPLC analysis revealed that the main compounds of XGLXJD decoction were paeoniflorin, isorhamnetin, quercetin, luteolin, kaempferol, and baicalein. The molecular docking assay indicated that the docking energies of the main compounds of XGLXJD decoction to the top hub genes were less than −5 kcal/mol. ELISA revealed that the administration of XGLXJD decoction decreased the levels of pro-inflammatory cytokines (TNF-α and IL-17). Furthermore, the AC, cAMP, and PKA levels were enhanced after its administration. IHC staining demonstrated that the administration of XGLXJD decoction activated the AC–cAMP–PKA–CREB signaling pathway in skin. In addition, it enhanced sucrose preference and forced swimming time percentage. It also enhanced the expression of cAMP and PKA in the hippocampus.

XGLXJD decoction alleviated psoriasis and depression-like behavior. The AC–cAMP–PKA–CREB signaling pathway may play a crucial role in mediating this effect.

There are many active substances in the composition of the Hypericum perforatum L. plant that have different and complex structures and show different pharmacological effects. It was accepted as a medicinal plant by the World Health Organization in 2002, and it is given a detailed place in the monographs under the name Hyperici herba. In these monographs, the anti-bacterial, anti-viral, anti-depressant, protein kinase-C inhibitor, and wound-healing effects of the plant were described.

The aim of this study is to determine the essential oil components in the samples of the H. perforatum L. plant, which is widely grown in the Thrace region in Türkiye.

H. perforatum L. plant growing naturally in the provinces of the Thrace region was collected, and the essential oils of the plant were obtained by using the hydro distillation method after the collected herbs were dried. Chemical content analyses were performed on a gas chromatography-flame ionization detection (GC-FID).

The main components of the essential oil of the plant are sesquiterpenes (ß-caryophyllene, caryophyllene oxide, β-selinene, α-selinene, β-farnesene, α-amorphene, spathulenol), alcohols (1-tetradecanol, 1-tridecanol, phytol, and alpha cadinol), and fatty acids (pentadecanoic acid, palmitic acid, and 9, 12 octadecadienoic acid).

This is the first study to determine the chemical composition of essential oil in H. perforatum L. samples collected from the Thrace region in Türkiye. The essential oil content of the plant can be affected by factors such as the developmental stages of the plant (preblooming, flowering stage, and fruiting time), use of fresh or dry plant material, extraction method, genetic parameters, and climate.

Rutin protects and stabilizes hepatocytes and is used to treat hepatitis, liver cirrhosis, and other diseases. However, there is little evidence to suggest that Rutin is associated with CCl₄-induced hepatotoxicity. In this study, we investigated the effects of Rutin on CCl₄-induced liver inflammation and discussed in vivo and in vitro mechanisms.

CCl₄ was used to induce liver inflammation, and Rutin was used for intervention in mice. Then the body weight, liver weight, and liver inflammation of all mice were measured. Western blotting and RT-PCR were used to assess TLR4/MyD88/NFκB inflammation signaling pathways and inflammatory gene expression. Finally, TLR4 was activated in primary hepatocytes to verify the above signal pathways.

CCl₄ mice developed liver function damage and liver inflammation. Further investigations showed that the inhibitory effects of Si on inflammation were mediated by TLR4/MyD88/NFκB inflammation signaling pathways inhibition. Primary hepatocyte studies also confirmed the participation of these signaling pathways and proteins.

Rutin improved CCl₄-induced liver inflammation. Such mechanisms may be related to TLR4/MyD88/NFκB inflammation signaling pathways.

Sanqi Xueshangning Capsules (SXC) is a famous Chinese patent medicine with a good curative effect on various gynecological pain diseases in China. However, its chemical composition and potential active ingredients were clearly unrevealed, which hindered the elaboration of its material basis functional mechanism and clinical application.

In the study, we developed an integrative ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) method coupled with SCIEX OS software for characterizing the chemical composition in SXC. The data collection was conducted using a Waters UPLCTM HSS T3 (2.1 × 100 mm, 1.8 µm) in a binary elution system. The SCIEX OS software coupled with multivariable processing techniques was used to screen absorbed prototypes in serum after oral administration of SXC. The structural identification of the components was performed by MS/MS fragmentation mechanism.

A total of 113 components from SXC were tentatively characterized, including 38 saponins, 13 flavonoids, 10 phenylpropanoids, 9 alkaloids, 20 organic acids, and 23 others. Based on the in vitro composition results, 21 absorbed prototypes in rat serum were identified, mainly including alkaloids, flavonoids, phenylpropanoids, and saponins.

It was concluded that an integrative UPLC-Q-TOF-MS method coupled with SCIEX OS software could be successfully applied for the components’ characterization of SXC in vitro and in vivo, which laid a reliable scientific basis for promoting the discovery of material basis and the improvement of quality control of SXC.

Prolactinoma is a common intracranial tumor with a high incidence and serious harm to human health. At present, there is only one therapeutic drug in China, bromocriptine, and the Chinese herb Prunella vulgaris L. (P. vulgaris)(PV) has shown certain anti-prolactinoma effects in the early stage. We expect to develop a candidate drug against prolactinoma.

First, the extracts of P. vulgaris L.(PVE)were extracted with water, and the cell proliferation test of rat pituitary tumor cells MMQ was checked by the Cell Counting Kit-8 (CCK-8) assay. Then, core targets and correlative pathways were selected by the “protein–protein interaction” (PPI) network and the “PV–Target–Prolactinoma” network. The core targets and main components simulate the binding by molecular docking. Finally, the PVE and MMQ cells were used to verify the results.

Through the CCK-8 assay, the PVE inhibited the proliferation of MMQ cells. From the network pharmacology, the 21 targets, 9 signaling pathways, and 20 gene ontology (GO) projects were attained (p < 0.05). As a result the estrogen receptor α (ESR1), RAC-alpha serine/threonine-protein kinase(AKT1), and mitogen-activated protein kinase 3(MAPK3)were the core targets of protein against prolactinomas, which was in line with western blotting analysis.

Our findings demonstrate that the PVE was verified against prolactinomas through the ESR1, MAPK3 targets, and the phosphoinositide 3-kinase/protein kinase B pathway.

Phytopathogenic fungi remain the main infectious agents in plants, causing severe damage to the environment and human health. Thus, to reduce the usage of synthetically derived fungicides and perform agricultural crop production, the search for new control strategies including plant extracts constitutes an eco-friendly and safe alternative.

This study aimed to quantify the phytochemical constituents of the three plant (Mentha pulegium L., Mentha spicata L., and Mentha longifolia L.) extracts and to screen their phytochemical composition including total phenolic (TPC), flavonoids (TFC) and condensed tannins contents (TCTC), and to evaluate their antioxidant activities. The efficacy of all mint extracts will be investigated against phytopathogenic fungal species.

The three plant extracts were screened to assess their total phenolic, flavonoids, and condensed tannin contents using spectrophotometric assays. The antioxidant activities include 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferric ion reducing antioxidant power (FRAP), and β-carotene assays. The antifungal activities were investigated on phytopathogenic species including Botrytis cinerea, Fusarium culmorum, Fusarium oxysporum, Aspergillus niger, Aspergillus flavus, and Trichoderma sp.

Quantitative analyses of phytochemical constituents of Mentha genus extracts revealed that both ethyl acetate (EtAc) and chloroformic (Chl) extracts are a rich source of phenols, flavonoids, and condensed tannins. Ethyl acetate extract of M. longifolia (EtAc L) displayed the highest content of phenols (69.9 ± 1.35 mg GAE/g DW) and flavonoids (53.26 ± 2.11 mg CE/g DW), while M. pulegium ethyl acetate extract (EtAc P) has the highest condensed tannins content (2.13 ± 0.4 mg CE/g DW). Moreover, the tested extracts exhibited potent antioxidant activities at low concentrations for EtAc L, followed by M. spicata (EtAc S), and EtAc P (IC₅₀ = 35.76 ± 1.32 µg/mL for scavenging DPPH free radicals; EC₅₀ 527.96 ± 5.45 µg/mL for FRAP, and IC₅₀ = 106.3 ± 3.75 µg/mL for β-carotene bleaching test). Finally, all tested extracts were able to inhibit the growth of several phytopathogenic micro-organisms on both agar and broth media.

The Mentha extracts derived from the three mint species (i.e., L, P, and S) could be used for their antifungal activities to provide sustainable crop pest management.

Osteoporosis is a chronic metabolic bone disease caused due to the dysregulation in the functioning of osteoblast and osteoclast cells leading to increased bone resorption predominantly in postmenopausal women and the elderly, thereby affecting a significant global population.

The present study aims to computationally screen the phytochemical constituents of Oldenlandia umbellata Linn against the unique molecular therapeutic targets of osteoporosis and validate the results using Molecular Dynamics (MD) Simulation.

The molecular docking analysis of the selected phytoconstituents against the molecular targets were performed using AutoDock 4.2.6 – AutoDock Tools 1.5.6. In addition, the drug likeliness, ADME, bioactivity, and toxicity were predicted using Molinspiration ADMELAB2.0, ProTox-II and Orisis DataWarrior online tools. Molecular Dynamic Simulation studies were performed using WebGRO macromolecular simulation server.

Molecular docking results and data analysis revealed that deacetylasperuloside possesses good drug-likeness, ADME properties, and bioactive scores and did not indicate any toxicity compared to other phytochemicals exhibiting binding energies below –8.00 Kcal/mol against the targets. Together, the study emphasized that deacetylasperuloside could bind with the molecular targets of osteoporosis, and the lead is a potential therapeutic candidate for osteoporosis treatment. Further, molecular dynamic simulation analysis for 100 ns revealed that the ligand–protein complexes, including glycogen synthase kinase 3β (GSK3β)-deacetylasperuloside and cathepsin K (CTSK)-deacetylasperuloside complexes, were stable and highly compact assessed from their trajectories. Hence, it can be emphasized that deacetylasperuloside could be a potential therapeutic molecule that could inhibit the targets, including GSK3β and CTSK.

Oldenlandia umbellata L. is a potential candidate for identifying therapeutic leads for osteoporosis treatment. Further, in vitro and in vivo studies are needed as an output of this research to evaluate its therapeutic efficacy.

This study investigated the effects of BP and doxorubicin (DOX) on 4T1 tumor cells in a mouse model.

After inducing breast tumors, 70 4T1-tumor-bearing BALB/c mice were divided into seven groups (n = 10/group). The groups were treated with DOX and BP for 35 days. On the 36th day, blood was taken from the heart, and serum was separated to measure levels of cytokines, estrogen, progesterone, testosterone, and nitric oxide. Antioxidant enzyme activities, as well as tissue ferric-reducing antioxidant power (FRAP) and malondialdehyde (MDA) levels, were evaluated. The expression of apoptotic genes and metastasis was measured using real-time polymerase chain reaction (PCR), while the expression of apoptotic proteins was evaluated using Western blotting. Finally, Ki-67 and p-53 were examined using immunohistochemistry to determine apoptosis.

The study found that BP, with its synergistic effects with DOX, reduced the volume of tumors and increased the expression of apoptotic genes and proteins. In a dose-dependent manner, groups receiving BP and DOX with their synergistic effects reduced the level of estrogen and nitric oxide and also reduced the level of pro-inflammatory cytokines. BP along with DOX increased serum interferon-γ (IFN-γ) levels. Tumor tissue FRAP and thiobarbituric acid reactive substances (TBARS) levels increased in BP-treated groups. Ki-67 and Bcl-2 proliferation markers levels decreased, and p53 levels increased in 4T1-breast tumors.

The study concluded that BP with its synergistic effects along with DOX has the ability to suppress the growth of tumors and can also inhibit the oxidative damage of DOX.

In India, cancer of the cervix is the second most common cancer in women. Nowadays, various traditional herbal formulations can be used as adjuncts during conventional cancer therapies due to their anticancer activity. Habbe Musaffi Khoon (HMK), a Unani compound formulation, is traditionally used as a blood purifier and thus used to treat boils, scabies, acne, and pimples. It is also indicated in skin diseases, Habis-ud-Dam (styptic), and therapeutically used in Nafz-ud-Dam (hemorrhagic).

The main purpose of this work was to phytochemically characterize HMK aqueous extract (HMKaq) and determine its activity against cervical cancer cells.

HMKaq was subjected to liquid chromatography-mass spectrometry (LC-MS) and evaluated for the presence of phytocompounds. HMKaq was evaluated for its effect on the viability of cervical cancer cells (HeLa, SiHa, and C33A) and was determined by MTT assay. Trypan blue dye exclusion was used to determine the outcome of HMKaq on the growth kinetics of cervical cancer cells. The role of HMK in regulating the cell cycle was analyzed by FACS and its effect on apoptosis was evaluated by checking mitochondrial membrane potential readings of JC-1 aggregates on a microplate reader and on the mRNA expression of caspase-3 and cytochrome C was evaluated by PCR.

LC-MS analysis revealed 3845 compounds, out of which 58 were the major compounds. These included phenolics, esters, fatty acids, furans, quinones, tannins, flavonoids, and terpenes. HMKaq reduced the viability and growth of the cells. It induced arrest in the cell cycle of HeLa and apoptosis in SiHa and C33A by depolarizing the membrane potential of the mitochondria and upregulation of mRNA expression of Cas3 and Cyt C.

HMK exhibited anticancer activity against cervical cancer cells, thereby signifying its therapeutic potential.

Zingiber zerumbet (L.) Smith (lempoyang) has a traditional application in the treatment of indigestion, worm infestation, loss of appetite, and postpartum conditions.

Extracts of Z. zerumbet (methanol, hexane, and ethyl acetate) were utilized to investigate the mutagenic, anti-mutagenic, and cytotoxic properties.

Initially, a mutagenicity test (Ames test) was conducted, followed by an anti-mutagenicity test, to assess the potential of Z. zerumbet extracts in inhibiting mutagenicity induced by sodium azide and 9-aminoacridine. Furthermore, the cytotoxic ability of the extracts was evaluated using the methyl thiazolyl tetrazolium (MTT) assay.

The mutagenicity assessment revealed that the methanol extract of Z. zerumbet exhibited a twofold increase in the number of revertants in Salmonella typhimurium strain TA 1537. Similar results were observed for the hexane extract, except at a concentration of 6.25 mg/mL, where no significant increase in revertants was observed. On the other hand, the ethyl acetate extract demonstrated a twofold increase in revertants in S. typhimurium strain TA 1535. Notably, the ethyl acetate extract displayed remarkable anti-mutagenic activity against 9-aminoacridine, while the hexane extract exhibited strong anti-mutagenic activity against sodium azide. Regarding cytotoxicity assessment using the MTT assay, the methanol extract of Z. zerumbet exhibited the highest cytotoxicity with an IC₅₀ value estimated at 388.50 ± 135.75 µg/mL. The hexane and ethyl acetate extracts showed IC₅₀ values of 514.17 ± 135.75 and 589.67 ± 241.67 µg/mL, respectively.

The extracts displayed both mutagenic and cytotoxic activities. However, they also exhibited promising anti-mutagenic potential, which could be harnessed for cancer prevention purposes.

Finding new effective substances to develop drugs against cancer or to prevent it remains a big challenge facing scientists and clinicians.

In such an issue, we tested the anti-tumoral effect of Scorzonera undulata extracts on MCF7 cells.

Methanolic extracts of S. undulata roots (RSU) and aerial parts (ASU) from the Ha’il region were analyzed by HPLC. Their DPPH scavenging, ferric-reducing antioxidant power (FRAP) and total antioxidant potentials were evaluated. The MTT test was used to determine their cytotoxic effects on MCF7 cells using different concentrations, and their potential to induce apoptosis was determined by measuring the mitochondrial transmembrane loss of potential by flow cytometry using the JC-1 fluorescent marker.

S. undulata was found to be rich in flavonoids, tannins, and polyphenols. ASU contains great amounts of apigenin and gallic acid, whereas RSU contains luteolin and chlorogenic acid as major components. The potential to scavenge DPPH and the total antioxidant activity of ASU were twofold greater than those of RSU. MCF7 cell viability was reduced by 50% at concentrations of 4.22 ± 0.06 and 5.89 ± 0.08 mg.mL⁻¹, respectively, for ASU and RSU at 24 h. In addition, S. undulata extracts induced MCF7 cells’ death through cell lysis and apoptosis.

As an edible vegetable, S. undulata is worth further exploration in alternative and complementary medication as it possesses significant antioxidant and anti-cancer potentials.

4-Methylumbelliferone (4-MU) is a coumarin compound that can be extracted from the medicinal plant with anti-cancer properties, Smilax china L. In recent years, studies have revealed its potential as an anti-tumor and anti-metastasis drug with promising effects in cancer treatment. Despite an increase in research on the metabolic patterns of tumor cells, no prior research has suggested that 4-MU inhibits tumor proliferation by blocking glycolysis. This thesis presents evidence that 4-MU binds to proteins involved in glycolysis, thus mediating its anti-tumor effects.

Network pharmacology, transcriptomics, and molecular docking were utilized to forecast the potential targets and probable pathways of 4-MU’s anti-cancer activity, and the affinity of 4-MU towards potential targets was discovered using microscale thermophoresis (MST) detection.

The results of transcriptome analysis brought to light that the genes with differential expressions were primarily enriched in metabolic pathways, including glycolysis-related proteins. Using network pharmacology and molecular docking, our study identified Heat Shock Protein 90 Alpha Family Class A Member 1 (Hsp90AA1), mitochondria, phosphoglycerate kinase 2 (PGK2), glycerol-3-phosphate dehydrogenase (GPD2), and glucose-6-phosphate isomerase (GPI) as potential targets of 4-MU. The strong binding affinity between 4-MU and these proteins was confirmed by the MST assay.

The findings indicate that 4-MU can hinder glycolysis by binding to glycolysis-associated proteins such as Hsp90AA1, PGK2, GPD2, and GPI. This results in the prevention of the energy supply to the tumor tissue, which ultimately curbs tumor growth, thereby demonstrating its anti-tumor properties. These results conclude that 4-MU has the capacity to be a novel glycolysis inhibitor for cancer treatment. Moreover, the identification of these glycolysis-associated proteins as possible targets for cancer therapy offers new avenues for research in the field of cancer treatment, thus providing further valuable evidence for the anti-cancer mechanism of 4-MU.

Ilexsaponin A₁ (IA₁) is a bioactive triterpene saponin derived from natural medicinal plants. IA₁ exhibits anti-inflammatory and proangiogenic activities and improves intestinal barrier function. It has been reported that IA₁ could be metabolized into a dominant metabolite, ilexgenin A (IA) by β-glucosidase enzymes in intestinal microflora.

Herein, an accurate, sensitive, and selective method based on ultra-performance liquid chromatography coupled with mass spectrometry was established to simultaneously profile the metabolism and pharmacokinetic behaviors of IA₁ in normal and antibiotic-treated rat plasma after intragastric administration of IA₁. The precursor-to-product ion pairs of IA and IA₁ were m/z 501.32↓439.32 and m/z 663.38↓501.32, respectively. For method validation, the specificity, matrix effect, accuracy, precision, and stability of the pharmacokinetic study were measured, and a calibration curve was created. The collaborative pharmacological target pathways of IA₁ and its metabolite IA were investigated using network pharmacology tools.

The validated analytical method was successfully utilized to investigate the pharmacokinetics of IA₁ in normal and antibiotic-treated rats. The bioavailability of IA₁ and conversion from IA₁ to IA were significantly inhibited by antibiotic-treated rats after oral administration of IA₁. Fragment ions at m/z 483.3155, 455.3159, 439.3233, 421.3136, 395.3362, 152.9952, 113.0256, and 71.0531 were characteristic of the IA₁ moiety. IA₁ was metabolized in rat plasma by biotransformation routes involving deglycosylation, decarboxylation, isomerization, hydrogenation, dehydrogenation, and oxidation. Considering database analysis, IA and IA₁ play synergistic role in common pharmacological pathways, such as hypertrophic cardiomyopathy and dilated cardiomyopathy.

The experiments illustrated that β-glucosidase activity inhibited by antibiotics suppressed the hydrolysis reaction of IA₁ in the intestinal tract. IA₁ and IA play a synergistic role in exerting effects.

Insulin resistance (IR) is a pathological state closely associated with various diseases, with hepatic insulin resistance playing a pivotal role. Insulin resistance can be improved by Ginsenoside Rg1 (GRg1), which is known as one of the most biologically active compounds found in ginseng. Nevertheless, the precise role and mechanisms of GRg1 in ameliorating hepatic insulin resistance are still unknown.

We wanted to demonstrate the impact of GRg1 on hepatic insulin resistance and explore the underlying mechanisms in our work.

We mimicked insulin resistance conditions by culturing HepG2 cells in 30 mM glucose for 24 h. The effects of GRg1 on cellular glucose consumption and the key kinases of the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT)/glycogen synthase kinase-3β (GSK-3β) pathway were evaluated. To assess if the PI3K/AKT/GSK-3β pathway is crucial for GRg1’s protective effect against insulin resistance, the compound LY294002, which inhibits PI3K, was employed.

In HepG2 cells, GRg1 markedly increased glucose uptake while exhibiting no cytotoxicity. Additionally, GRg1 activated the PI3K/AKT/GSK-3β pathway, as indicated by the increased phosphorylation of insulin receptor substrates-1 (IRS-1), AKT, and GSK-3β. Treatment with LY294002 significantly reversed the promotive effects of GRg1 on cellular glucose consumption and PI3K/AKT/GSK-3β pathway activation.

Taken together, our present study revealed GRg1 exerted a protective effect against hepatic insulin resistance induced by high glucose through PI3K/AKT/GSK-3β pathway, suggesting its potential as a beneficial therapeutic medication.

Asthma is a prominent non-communicable inflammatory disease that affects both children and the elderly. Younger people are more prone to asthma, and most prescribed anti-asthmatic medicines relieve symptoms but do not cure the condition completely. We investigated the ability of a phytochemical plumbagin to alleviate ovalbumin (OVA)-induced asthma in BALB/c mice.

The allergic asthma-induced mice were treated with two different doses of 25 and 50 mg/kg bwt plumbagin, and to compare the efficacy of plumbagin, a standard drug dexamethasone treatment was given. OVA-specific IgE and eotaxin were quantified to determine the induction of asthma and the inhibitory role of plumbagin. Total leukocyte and differential count were done to assess the effect of plumbagin on inflammatory cells. Inflammatory cytokines inducing both atopic and non-atopic asthma were quantified to examine the efficacy of plumbagin against allergic and non-allergic-induced asthma. Nitric oxide (NO) and myeloperoxidase activity were measured to investigate the anti-asthmatic potential of plumbagin. The antioxidant potency of plumbagin was assessed by quantifying the levels of antioxidants and the oxidative stress marker malondialdehyde. Lung weight index and histopathological analysis of lung tissue were done to confirm the ameliorative potency of plumbagin against OVA allergen-induced asthma.

Plumbagin treatment significantly decreased the status of OVA-specific IgE and eotaxin, thereby prevented the eosinophilic infiltration. It also inhibited the synthesis of both atopic and non-atopic inducing inflammatory cytokines. Plumbagin treatment also increased the levels of antioxidants and prevented the lung tissue damage, which was evidenced with our histopathology study of lung tissue.

Overall, our finding confirms that plumbagin is persuasively alleviated OVA allergen-induced asthma complications in mice model and may be an alternative for currently available anti-asthmatic drugs.

Adverse effects of benzene (BZ) exposure toxicities were documented by many researchers worldwide. BZ exposure causes many hematological abnormalities that might be treated with naturally occurring phytocompounds.

In this experimental study, we evaluated the anti-inflammatory effect of L-carvone and thymoquinone together (LCTQ) against BZ-induced inflammatory toxicities in Sprague-Dawley (SD) rats.

BZ exposure could cause an excess formation of immature blood cells to enter the peripheral bloodstream. Phytocompounds namely L-carvone (LC) and thymoquinone (TQ) have confirmed anti-inflammatory effects against various diseases.

Rats were divided into four different groups such as control, LCTQ group, BZ pathologic group, and treatment group (LCTQ+BZ). After 10 weeks of the experimental period, body weight changes, hematological parameters, pro-inflammatory cytokines, oxidative stress, RBC antioxidants, bone marrow cellular abnormalities, and bone marrow DNA fragmentation were evaluated.

BZ toxicity showed abnormal loss of body weight, altered hematological parameters, increased pro-inflammatory cytokines, abnormal cellular oxidative status, and DNA damage. LCTQ treatment showed significant (p < 0.05) increase in body weight, normalized hematological parameters such as red blood cells, hemoglobin, white blood cells (lymphocytes and eosinophils), platelets (PLT), and hematocrit with RBC parameters, reduction of pro-inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6), controlled oxidative stress, normalized enzymatic antioxidants in the RBC cells, normalized nucleated cells, megakaryocyte cells, and controlled DNA fragmentation were observed.

The current study showed an anti-inflammatory effect of LCTQ through the control of inflammation against benzene-induced toxicities in rats.

Previous research has highlighted the regulatory role of miR-182-5p in targeting TLR4 during the pathogenesis of allergic rhinitis. In a different context, TLR4 has been identified as a crucial factor in the development of lung ischemia-reperfusion injury, where its upregulation is believed to initiate the injury process. Additionally, miR-199a-3p has been shown to possess cardioprotective properties in simulated ischemia/reperfusion (I/R) injury models.

HE and TUNEL were performed to evaluate the cardiac injury and cellular apoptosis of I/R mice under distinct conditions. Real-time PCR was used to analyze the expression of microRNAs (miRNAs) and mRNAs under differential conditions.

Pretreatment by Shexiang Tongxin Dropping (SXTXD) has been shown to significantly augment the therapeutic efficacy of MSC-derived exosomes (MSC-EXOs) in attenuating cardiac injury in I/R mice. MSC-EXOs effectively restored the repressed miR-182/miR-199a-3p expressions and activated TLR4/CD44 expressions in I/R mice, while SXTXD pretreatment remarkably strengthened the efficiency of MSC-EXOs. Moreover, SXTXD pretreatment notably reinforced the capability of MSC-EXOs to maintain cardiac parameters, including iNOS and interleukin-1β (IL-1β). Furthermore, the luciferase assay indicated that miR-182-5p and miR-199a-3p effectively suppressed the luciferase activities of TLR4 and CD44, respectively, through binding to the 3´ UTR. The overexpression of miR-182-5p and miR-199a-3p significantly suppressed the expression of TLR4 and CD44 in H2C6 and RAW264.7 cells.

In conclusion, our investigation indicates that MSC-derived exosomes, pretreated with SXTXD, hold promise in mitigating cardiac I/R injury by modulating inflammatory responses through the miR-182-5p/TLR4 axis and miR-199a-3p/CD44 axis. These findings suggest potential therapeutic strategies for addressing I/R-related cardiac complications.

Extracts of Chenopodium hybridum L. leaves and stems exerted a significant anti-proliferative effect on human A2780 ovarian cancer cells, but C. hybridum active components have not been reported.

Here, a method is described for screening of C. hybridum extracts for potential bioactive components that inhibit A2780 cell proliferation. First, the spectrum–effect relationship between UPLC-Q-Exactive MS chromatograms and C. hybridum extract antiproliferative effect against A2780 cells was established to evaluate extract bioactive components using partial least squares (PLS) analysis.

The results indicated that the optimal reflux extraction process for preparing C. hybridum extracts with antiproliferative activity involved a suspension of C. hybridum material in 8 volumes of 70% ethanol followed by heating and refluxing twice for 60 min/reflux step and then repeating the extraction and pooling of both the extracts. Chromatographic results revealed five compounds with potential anti-ovarian cancer activities based on inhibition of A2780 cell proliferation: isorhamnetin-3-O-β-D-furanosyl(1→2)-O-[α-L-rhamnpyranosyl(1→6)]-β-D-glucopyranoside, kaempferol-3-O-β-D-glucopyranoside-7-O-α-L-pyranoside, kaempferol-3-O-[α-L-rhamnopyranosyl(1″→2″)]-β-D-galactopyranoside, quercetin-3,7-di-rhamnose, and isorhamnetin-3-acacia disaccharide. Network pharmacological screening revealed nine core cellular targets that potentially interacted with these compounds.

These results were verified through molecular docking studies that supported the involvement of these compounds in observed C. hybridum A2780 cell antiproliferative effects, thus indicating C. hybridum active components may have value in ovarian cancer treatments.

Several studies have demonstrated the anti-inflammatory properties of gastrodin. However, its specific role and mechanism in the context of osteoarthritis (OA) remain unclear.

The objective of this study was to explore the function and mechanism of gastrodin in both in vivo and in vitro OA models.

In this study, the targets of gastrodin against osteoarthritis were analyzed using bioinformatics methods. The effect of gastrodin on neutrophil infiltration and inflammatory cytokines in OA synovial tissue was evaluated using H&E staining. The levels of inflammatory cytokines were determined using ELISA and Q-PCR. The extracellular matrix degradation-related proteins and PI3K/AKT/FoxO1 signaling pathway were tested using western blotting.

Bioinformatics analysis predicted that the target of gastrodin against OA might act on the FoxO pathway. The use of gastrodin resulted in a significant reduction in the infiltration of neutrophils in the synovial tissue of rats, as observed through H&E staining. Additionally, gastrodin was found to attenuate IL-1β-induced inhibition of chondrocyte viability, pro-inflammatory cytokine production, and extracellular matrix degradation. This effect can be attributed to the suppression of PI3K/AKT/FoxO1 by gastrodin.

These findings indicate that gastrodin is a treatment for OA.

In the study, we investigated the effects of an Eriocheir sinensis water extract on a reflux esophagitis-induced rat model.

The crab has been used as a traditional medicine and food source in many countries worldwide for a long time, and the crab contains various useful substances such as chitin, protein, calcium carbonate, etc.

All rats were fasted for 24 h and then orally pretreated with saline, Eriocheir sinensis (100 mg/kg), or ranitidine (40 mg/kg). Reflux esophagitis was induced by pylorus and forestomach ligation. Morphological changes in the mucosal damage in the esophagus were analyzed by the ImageJ program. Also, the expression of the inflammatory proteins and cytokine in the esophagus was measured by western blot assay to demonstrate the protective effects of Eriocheir sinensis in reflux esophagitis model. Histological analysis of esophagus was performed by hematoxylin & eosin (H&E) staining.

In the reflux esophagitis induction group, the esophageal tissue damage caused by the induction of reflux was 60% of the whole. Eriocheir sinensis administration significantly ameliorated esophageal mucosal damage by 40%, also determined by histological evaluation of reflux esophagitis in rats. Eriocheir sinensis was also found to downregulate the expression levels of proteins such as cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), and tight junction protein (claudin-5) compared to the reflux esophagitis induction group. In addition, Eriocheir sinensis markedly attenuated the activation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and the phosphorylation of the inhibitor of NF-κB (IκBα).

These results indicated that Eriocheir sinensis suppressed the development of esophagitis and modulated inflammation by regulating NF-κB activation. Based on these findings, we concluded that Eriocheir sinensis can protect the esophageal mucosa from reflux esophagitis.

Osteosarcoma (OS) is a highly metastatic primary bone malignancy, and the treatment options remain inadequate. Hence, exploring innovative natural medications is required. Prunetin (PRU) is an isoflavone that has been a proven anticancer agent in numerous cancer cell lines. However, the activity of PRU against OS remains uncertain.

Here, we studied the anticancer activity of PRU (20 and 25 µM) on human OS cells MG-63 and investigated its latent mechanism. The PRU activity of MG-63 cells cytotoxicity, intracellular ROS, metastasis, apoptosis, anti-apoptotic proteins, MAPK/STAT-3, and AKT signaling pathways was assessed by MTT assay, DCFH-DA, DAPI, PI, AO/EB, cell adhesion, and RT-PCR analysis. Findings unveiled that PRU could constrain MG-63 cell viability and adhesion through elevated intracellular ROS and elicited apoptosis.

Likewise, PRU (20 and 25 µM) avert the MG-63 cell proliferation, which stimulates apoptosis by the enhancement of Bax and caspases, while it diminishes Bcl-2 in a dose-dependent way. Furthermore, PRU could reduce Pin-1, and anti-apoptotic elements, as well as trigger apoptotic signaling pathways. Our data established that PRU alleviates MG-63 cell proliferation and metastasis via ROS-mediated apoptosis, which triggers MAPKs/STAT3 and AKT pathways, suggesting that PRU is a promising natural remedy for OS. In order to comprehend the therapeutic target for cancer, we assessed the effect of PRU on the expression of Pin1, which is thought to be over-expressed in many human malignancies. According to our findings, PRU specifically suppressed Pin1 expression to reduce the expression of Akt, STAT3, P38, JNK, P65, and IL-6. We evaluated the impact of PRU on the expression of Pin1, which is allegedly over-expressed in many human malignancies, to better understand the therapeutic target for cancer. Researchers state that PRU inhibited the expression of Akt, STAT3, P38, JNK, P65, and IL-6 in particular, by suppressing Pin1 expression.

Together, these results suggest that PRU may be an effective treatment for bone cancer in people by preventing Pin1 expression.

Epilepsy is a chronic neurological condition with various underlying mechanisms. It is known to affect a multitude of people across the globe, regardless of age and gender. Seizures associated with epilepsy involve the participation of stimulatory glutamatergic mechanisms along with inflammation and oxidative damage.

In this investigation, the anti-epileptic effect of scutellarein, a phytochemical compound isolated from Erigeron breviscapus (Vant.), has been evaluated in the pentylenetetrazol (PTZ) kindling epilepsy model in mice.

The experimental mice were categorized into six groups with six animals in each. The first control group was given normal saline. The second group was provided only PTZ through an intraperitoneal route to induce seizures. The third and fourth groups received two oral doses of scutellarein (10 and 20 mg/kg) before 30 min of PTZ induction. Diazepam was intraperitoneally administered to the fifth group as a positive control. The impact of scutellarein on the duration and initiation of clonic and tonic convulsion, mortality, kindling, mobility, and immobility duration in PTZ-induced rodents was estimated. Also, the impact of scutellarein on oxidative stress markers and antioxidant and inflammatory marker levels was also evaluated.

Scutellarein treatment was able to reduce PTZ-induced seizures in mice. In PTZ animals, scutellarein lowered the seizure severity by suppressing the onset and duration of convulsions. Scutellarein successfully modulated the PTZ-provoked changes in gamma-aminobutyric acid (GABA), glutamate, and dopamine levels, as well as Ca²⁺ ATPase and Na⁺ K⁺ ATPase activity.

Furthermore, it remarkably reduced the oxidative stress markers and decreased the contents of nuclear factor-kappa B (NF-κB), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) in PTZ animal brain tissues, confirming its anti-convulsant potential.