European Journal of Inflammation

Serine protease inhibitors (serpins) are a superfamily of proteins involved in many important biological processes, including inflammation. Serpins dysfunction-related diseases are mainly treated by augmentation therapy using serpins purified from human plasma. Pnserpin from hyperthermophilic archaeon Pyrobaculum neutrophilum showed protease inhibition activity and high stability. In this study, we examined the anti-inflammatory activity of Pnserpin using xylene-induced acute inflammatory model of mouse ear swelling and lipopolysaccharide (LPS)-induced murine RAW 264.7 macrophages cellular model. The inhibition of mouse ear swelling and the production of pro-inflammatory cytokines in mouse serum or in macrophages cell were used to evaluate the anti-inflammatory effect of Pnserpin. Our results showed that Pnserpin could inhibit the xylene-induced mouse ear swelling and suppress the production of pro-inflammatory cytokines in mouse serum and in LPS-induced RAW264.7 cells. This study indicated that Pnserpin might have anti-inflammatory effect in vivo and in vitro.

Bronchiolitis obliterans (BO) is in general a rare and life-threatening form of non-reversible obstructive lung disease in which the bronchioles are compressed and narrowed by fibrosis and/or inflammation. The purpose of this study was to evaluate the clinical features of BO in pediatric patients and explore its risk factors. The medical records of 35 pediatric patients with BO at Guangzhou Women and Children’s Medical Center were evaluated. The age at onset of symptoms was 2–42 months (mean 13.3 ± 8.9 months), with age at diagnosis of 5 months–4 years (mean 17.8 ± 9.0 months). High-resolution computed tomography findings included mosaic pattern (100%), atelectasis (37.1%), air trapping (31.4%), and bronchiectasis (20.0%). Three patients received lung biopsies and mainly exhibited an inflammatory process surrounding the lumen of bronchioles. BO predominantly resulted from post-infectious causes (91.4%) which were primarily caused by adenovirus (50%), followed by Mycoplasma pneumoniae (46.7%) and influenza (20%). Pulmonary function tests (PFTs) showed severe and fixed airflow obstruction, decreased compliance, and increased resistance. No significant difference was found between before and after steroid treatment (P > 0.05). Two patients died owing to severe pulmonary complications, one of whom had inherent immunodeficiency. Our study suggests that the occurrence of BO, especially post-infectious BO, in China is relatively high and might result from primary immunodeficiency diseases in severe cases. Recurrent aspiration pneumonia caused by congenital dysplasia of the larynx and vaccination not on schedule might be potential risk factors for persistent and recurrent BO.

Mitochondrial dysfunction is involved in the process of sepsis and leads to the accumulation of reactive oxygen species (ROS), which breaks cellular homeostasis and activates the downstream inflammatory cascade. The autophagic removal of ROS is a well-established cellular adaptive mechanism. Adiponectin is an adipocytokine that plays an important role in metabolic and inflammatory regulation. In this study, we investigated the anti-inflammatory effect of adiponectin in a sepsis model and its potential association with autophagy. We induced RAW 264.7 macrophages with lipopolysaccharide (LPS) to set up the sepsis model and treated them with adiponectin, an inhibitor of the nucleotide-binding domain and leucine-rich repeat containing family pyrin domain–containing 3 (NLRP3), ROS, Complex I, and an autophagy inhibitor. Flow cytometry and western blot analysis were performed to detect the expression levels of ROS, NLRP3, interleukin-1 beta (IL-1β), microtubule-associated protein 1A/1B-light chain 3II/I (LC3II/I), and adenosine monophosphate–activated protein kinase (AMPK). Expression levels of NLRP3, IL-1β, and ROS were significantly increased following LPS induction, and adiponectin reversed this up-regulation. Meanwhile, adiponectin also enhanced the expression of LC3II/I, an autophagosome marker, but an autophagy inhibitor and AMPK inhibitor depleted (reversed) the anti-inflammatory and antioxidant effect of adiponectin. Taken together, in the LPS-induced sepsis model, adiponectin alleviated the inflammatory reaction by reducing ROS production, possibly by enhancing autophagy via the AMPK pathway. The activation of autophagy may therefore be a key mechanism by which adiponectin ameliorates the inflammatory reactions of sepsis.

Ginger (Zingiber officinale Roscoe) is one of the most commonly used medicinal plants and is extensively used for the treatment of arthritic patients in Traditional Korean Medicine (TKM) due to its various pharmacological properties. In this study, we evaluated the therapeutic effects of ginger on rheumatoid arthritis (RA), particularly focusing on the regulation of Th1, Th2, and Th17 cytokines and the inhibition of matrix metalloproteinase (MMP) release in mice with collagen-induced arthritis (CIA) and primary synovial fibroblasts. RA was induced in male DBA/1J mice via immunization with type II collagen (CII). A ginger extract was prepared in water. The ginger extract (100 and 200 mg/kg) or Mobic (50 mg/kg), as a reference drug, was orally administered to CIA mice once daily for 14 days after arthritis induction. Primary fibroblasts were isolated from the synovial tissues of osteoarthritis patients and then were stimulated with IL-1β and treated with the ginger extract at different concentrations. IL-4, IFN- γ, and IL-17 levels were measured in the serum or spleen and paw tissues of CIA mice and culture media via enzyme-linked immunosorbent assay (ELISA). The mRNA expression of IL-17, MMP-1, MMP-3, and MMP-13 was also detected in paw tissues and synovial fibroblasts through reverse transcription polymerase chain reaction (RT-PCR). Histological changes in the knee joints were observed via hematoxylin and eosin (H&E) and safranin-O staining. The major compounds in the ginger extract were analyzed using high-performance liquid chromatography (HPLC). Treatment with the ginger extract at 100 or 200 mg/kg significantly decreased the levels of IL-4, IFN-γ, and IL-17 and inhibited the expression of IL-17 in the spleen and paw tissues of CIA mice. Ginger extract inhibited the expression of MMP-1, MMP-3, and MMP-13 in the paw tissues of CIA mice and reduced inflammatory bone destruction in joint tissues. In IL-1β-stimulated synovial fibroblasts, the ginger extract significantly decreased the production of IFN-γ and IL-17 via inhibition of mRNA expression. The ginger extract also suppressed the expression of MMP-1, MMP-3, and MMP-13 mRNA. Vanillylacetone, 6-gingerol, 6-shogaol, and 1,4-cineol were identified as the main compounds in the ginger extract. These results indicate that ginger can prevent RA progression by inhibiting the secretion of Th1/Th2 and Th17 cytokines and MMPs, which are involved in the pathogenesis of RA.

Sepsis, which could lead to mitochondrial dysfunction and cellular energy loss, always induces acute liver injury and has a high mortality rate. Tetramethylpyrazine (TMP) is an active extract from the Chinese herb Ligusticum chuanxiong and exhibits anti-sepsis activity. In this study, a rat sepsis model was first established via cecal ligation and puncture (CLP). Then, 48 Sprague Dawley male rats were randomly divided into four groups (12 rats in each group): control group (C), sepsis group (S), TMP treatment group (T), and TMP prevention group (P). Serum aspartate aminotransferase (AST), serum alanine aminotransferase (ALT), mitochondrial aspartate aminotransferase (mAST), and adenosine triphosphate (ATP) levels and mitochondrial membrane potential (MMP) were measured and used as indicators of hepatic dysfunction severity and mitochondrial function. In addition, the activities of Na⁺-K⁺-ATPase, Mg²⁺-ATPase, Ca²⁺-ATPase, and Ca²⁺-Mg²⁺-ATPase in the mitochondrial membrane, the expression level of AQP8 and some inflammatory factors, and the level of oxidative stress were measured to explore potential mechanisms. We found that AQP8 accepts signals from inflammatory factors upon stimulation and during various infections, and low AQP8 expression levels could result in further downstream mitochondrial dysfunction. In conclusion, our data demonstrated that TMP could ameliorate hepatocellular mitochondrial dysfunction by decreasing the inflammatory response and increasing AQP8 protein expression.

The aim of this study was to investigate the mechanisms of combination treatment with FK506 and natamycin on alleviating damage of the cornea in mouse model of fungal keratitis.

In this study, the mouse model of fungal keratitis was created by intrastromal injection with Fusarium solani or Aspergillus flavus. The mice received 5% natamycin eye drops 6–8 times a day, or the mice received 0.05% FK506 eye drops 2 times per day for 21 consecutive days. Corneal damage was evaluated by H&E staining. The protein expression levels of NLRP3 were detected by immunohistochemistry. Moreover, the markers of inflammasome activation including NLRP3, ASC, caspase-1, IL-1β, and IL-18 were detected by western blot.

Histopathological results showed increased corneal thickening, dense inflammatory cell infiltration, and loss of epithelial continuity in the corneas after fungal infection. In addition, NLRP3 positive signals were observed to be obviously increased in the corneas after A. flavus or F. solani infection compared to the control group. Furthermore, the NLRP3 inflammasome is induced by fungal infection, as evidenced by increased protein expression levels of NLRP3, ASC, caspase-1, and downstream cytokines, such as interleukin (IL)-1β and IL-18. However, the corneal damage was alleviated and the activation of the NLRP3 inflammasome was significantly inhibited by drug treatment. Besides, the treatment outcomes were better in combined treatment group than that in single-agent treatment group.

In conclusion, FK506 combined with natamycin alleviate fungi-induced corneal damage by suppressing NLRP3 inflammasome activation.

Several studies have demonstrated the neuroprotective effect of enriched environment (EE) and its positive effect on cognitive performance in pathological conditions, such as neurodegenerative diseases, epilepsy, and traumatic brain injury. However, the immunomodulatory effect of EE in normal rodents is not well characterized. To assess the immunomodulatory effect of EE, we randomly assigned normal mice to EE housing or standard environmental (SE) housing for 3 weeks. Behavioral alterations were evaluated by open field, fear conditioning, and Morris water maze tests. Immunohistochemical staining was performed to assess the expression of behavioral-related proteins, and enzyme-linked immunosorbent assay (ELISA) for brain-derived neurotrophic factor (BDNF), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) was also performed. We also measured the levels of RIP1 and RIP3 proteins using western blotting. EE significantly improved the cognitive performance which was associated with the increased expressions of BDNF, ionized calcium-binding adapter molecule 1 (Iba1), and glial fibrillary acidic protein (GFAP); EE did not influence any morphological changes in the brain tissue in adult mice; however, increased resistance to inflammation induced by TNF-α was observed. These findings indicate that EE can positively influence cognitive and behavioral performance in healthy adult mice by exerting environ-immuno effect on neural function.

Atherosclerosis is a chronic inflammatory disease in the vessel wall. As a representative inflammatory cytokine, C-reactive protein (CRP) participates in the formation and development of atherosclerosis. It is demonstrated that aldosterone induces CRP generation in vascular smooth muscle cells (VSMCs). This study explored the inhibitory effect of eplerenone on aldosterone-induced CRP expression in VSMCs and mechanism. In the in vitro experiments, rat VSMCs were cultured and aldosterone (10 nM) was used as a stimulant for CRP generation. VSMCs were pretreated with eplerenone for 1 h prior to the stimulation. Messenger RNA (mRNA) and protein expression were identified by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot, respectively. The production of reactive oxygen species (ROS) was observed by a fluorescence microscope. In the in vivo experiment, a model of hyperaldosteronism was established by the subcutaneous administration of aldosterone to rats with the osmotic minipumps for 4 weeks. Serum aldosterone and CRP levels were determined with a radioimmunoassay and ELISA (enzyme-linked immunosorbent assay), respectively. The results showed that eplerenone inhibited aldosterone-induced mRNA and protein expression of CRP in VSMCs in vitro and in vivo, and decreased the circulating CRP level of hyperaldosteronism rats. Meanwhile, eplerenone reduced aldosterone-stimulated ROS generation and aldosterone-activated ERK1/2 phosphorylation in VSMCs. In summary, eplerenone inhibits aldosterone-induced CRP generation in VSMCs by regulating the MR-ROS-ERK1/2 signal pathway. These results provide new evidence for the potential anti-inflammatory effect of eplerenone.

Relationships between inflammation and innate immunity in cancer are widely accepted today; however, the precise cell mechanisms mediating these relationships have not yet been elucidated. Interleukin (IL)-17 is a proinflammatory cytokine that has been reported to induce inflammation in patients with autoimmune diseases. Myeloid-derived suppressor cells (MDSC) may contribute to the negative regulation of immune responses during cancer and inflammation. Vascular endothelial growth factor (VEGF) is reported to have multiple biological actions including increasing vascular permeability, neovascularization, and possible inhibition of immune function in malignant diseases. This study investigated the status of systemic inflammation and immune suppression associated with IL-17 and VEGF in patients with breast cancer. IL-17 production and the serum levels of VEGF were also increased in advanced stages of the disease. The production of IL-12, which induces Th1 cells, and the stimulation index (SI), which is a marker of cell-mediated immune function, were both shown to decrease along with disease advancement. Also, the production of IL-17 and the VEGF levels were both positively correlated with the levels of MDSC, the neutrophil-to-lymphocyte ratio (NLR), and C-reactive protein (CRP), and were inversely correlated with IL-12 production and the SI. Nutritional markers, including prealbumin (PA), transferrin (TF), and retinol-binding protein (RBP), were also shown to be significantly lower in patients with high production of IL-17 or high levels of VEGF. These data clearly showed that IL-17 and VEGF, whose levels correlated with each other and with those of MDSC, were significantly associated with disease advancement, systemic inflammation, suppression of cell-mediated immunity including Th1 induction, and malnutrition.

Adhesive capsulitis, commonly described as “frozen shoulder,” is experienced as chronic soreness and discomfort accompanied by an increasing inability to use and manipulate the joint, which can cause disability and interfere with normal activities. The onset of the condition is sudden and the origins obscure. The more we know about how this condition arises and evolves in the human body, the better the medical profession can provide effective management and amelioration. In examining the pathology and pathophysiology of adhesive capsulitis, we found recent evidence that intra-articular use of hyaluronan can provide effective relief. In this letter, we summarized this research to obtain a better understanding of the pathophysiology of adhesive capsulitis and the effects of hyaluronan.

Obesity escalates at an alarming rate worldwide. It is associated with chronic low-grade inflammation, which is implicated in the pathogenesis of type 2 diabetes and cardiovascular diseases. Adipose tissue is a primary site of obesity-induced pro-inflammatory factors such as leptin. Curcuminoids are potent anti-inflammatory agents; however, evidence on the polar fraction of Curcuma longa is scarce and the effect on leptin release has never been investigated. Therefore, we will investigate the influence of aqueous C. longa extract on leptin release from human subcutaneous adipose tissue (SAT). To achieve this, SAT explants were obtained from patients who underwent abdominal surgery. Patients were both males (67%) and females (33%). Their average body mass index (BMI) and age were 33.4 kg/m² and 43 years, respectively. Tissue explants were treated in triplicate with or without 0.1 and 1 mg/mL aqueous C. longa extracts and incubated for 24 and 48 h. Enzyme-linked immunosorbent assay (ELISA) was used to determine the concentration of leptin secreted by SAT. We have shown that 24 h treatment of SAT with 0.1 and 1 mg/mL of C. longa polar extract inhibited the release of leptin, while 48 h treatment of SAT with only 1 mg/mL of C. longa polar extract inhibited leptin release. Concentration of basal leptin released from SAT derived from females was higher than from males. However, this could be because females were morbidly obese. Our data demonstrate for the first time the inhibitory effect of aqueous C. longa extract on leptin release, shedding light on the role of leptin and adipose tissue in the mechanism of C. longa in reducing low-grade inflammation. This avoids the poor solubility and consequently the low bioavailability disadvantages of curcuminoids.

RNF11 (RING finger protein 11) has been indicated to be related to numerous immunological diseases. As a neo-regulator in innate immunity, RNF11 was proved to inhibit antiviral signaling and interferon-β (IFN-β) production in human cell line. Chicken antiviral signaling pathway is different with other species. However, there is little study about the role of RNF11 in chicken antiviral signaling pathway and inflammatory response. This study was designed to identify the function of RNF11 in chicken antiviral signaling pathway and inflammatory response. We overexpressed and interfered RNF11 in chicken embryo fibroblasts (CEFs) and then used quantitative reverse transcription polymerase chain reaction (qRT-PCR) to investigate the poly (I:C)-induced production of IFN-β and inflammatory response factors IL2 and IL10. The results indicated that RNF11 extremely significantly decreased the production of IFN-β and highly significantly increased the messenger RNA (mRNA) levels of IL2 and IL10, but depletion of RNF11 obtained inverse results. Thus, we concluded that RNF11 not only can inhibit antiviral signaling but also plays an important role in inflammatory response in chicken.

The phenomenon that papillary thyroid carcinoma (PTC) is often accompanied by Hashimoto’s thyroiditis (HT) raised interest in further study of immune changes caused by thyroid autoimmunity. We aimed to characterize cytokine profiles in the peripheral blood of patients with PTC with or without HT, compared with nodular goiter (NG) and healthy control (HC) patients, in order to determine the autoimmunity-related differences among these groups. A total of 50 PTC patients were divided into two groups, according to whether or not concurrent HT existed. A total of 20 NG patients and 20 HC subjects were also included as controls. All PTC patients and NG patients who underwent surgical thyroidectomy and pathology examination were included. The serum levels of four cytokines, including interferon gamma (IFN-γ), interleukin (IL)-17, IL-10, and IL-35 were measured using AimPlex bead–based immunoassays. Peripheral IFN-γ and IL-35 increased significantly in PTC patients with concurrent HT. IL-10 and IL-35 increased significantly in three groups (PTC with or without HT and NG patients), while IFN-γ increased only in the two PTC groups. IL-17 was highest in the HC group, but significantly lower in the other three groups. IL-17, IL-10, and IL-35 may be involved in tumor onset and progression. Moreover, IFN-γ and IL-35 may play vital roles in the concurrence of PTC and HT, which could warrant further exploration.

The myeloid-derived suppressor cells (MDSCs) are important tumor-induced immune suppression cells which have gained great attention in recent years. In this study, the levels of MDSCs in peripheral blood of cervical carcinoma (CC) patients, cervical intraepithelial neoplastic (CIN) patients, and healthy donors were analyzed by flow cytometry. It was shown that the proportion of circulating MDSCs was elevated in CC and CIN patients compared to that in healthy donor (HD). Further analyses revealed that the percentage of MDSCs is increased along with the stage of CC, but has no difference among other clinicopathological factors, including age, tumor grade, lymph node invasion, and histopathology. Taken together, our results suggested that MDSC levels are indicative of suppressive functions within the tumor environment of CC. Therefore, large validation studies could help to develop new immunotherapeutic strategies for CC patients by inhibiting or eliminating MDSC functions.

This study aims to investigate the clinical characteristics of tracheobronchial foreign bodies (TFBs) in children and its methods of treatment, providing a theoretical basis for including TFBs into the clinical pathway for children and conducting bronchoscopy performed by senior residents and attending physicians under general anesthesia. The clinical data of 1060 patients diagnosed with TFBs from January 2015 to January 2016 were evaluated. The age, gender, clinical manifestations, foreign body properties, thoracic CT, and three-dimensional reconstruction, preoperative and postoperative complications, hospital stay, surgical and general anesthesia bronchoscopy, and foreign body removal surgery of these patients were analyzed. TFBs frequently occurred in 0- to 3-year-old patients, accounting for 92.5%, and 64.3% of these patients were male. There is no evident difference in foreign bodies detected in the left and right bronchus. Foreign bodies are mainly botanic, accounting for 88.9%, among which peanuts and melon seeds were mostly observed. All pediatric patients received tracheobronchoscopy under general anesthesia, and 97.3% of these surgeries were performed by senior residents and attending physicians. No complication or death occurred after the surgery. TFBs can be treated according to the clinical pathway. The timely and accurate diagnosis of TFBs and its performance under general anesthesia can evidently reduce the mortality rate. Senior residents and attending physicians can be qualified to perform the bronchoscopy after training.