Dopamine (DA) is a free radical scavenger that attenuates apoptosis. We studied the effects of normal saline (NS) and DA on diaphragm apoptotic protein expression following 60 min of inspiratory resistance loading in rats. We tested for 27 apoptotic-related proteins and found 12 in the diaphragm. Of the 12 proteins, superoxide dismutase copper zinc (SOD [CuZn]) and proprioceptive event related potential (PERP) were significantly higher in the DA group than in the NS and sham groups (p = .002, p = .007). DA group diaphragms had significantly greater expression of SOD (CuZn) than the NS (p = .005) and sham group diaphragms (p = .003). Likewise, the DA group had significantly greater expression of PERP than the NS group (p = .008). These results suggest that DA decreases diaphragm apoptosis through elevated expression of SOD (CuZn). The identification of 12 apoptotic-related proteins will assist investigators as they study diaphragm apoptosis.
Callahan, L.A., Nethery, D., Stofan, D., DiMarco, A., & Supinski, G. (2001). Free radical-induced contractile protein dysfunction in endotoxin-induced sepsis. American Journal of Respiratory Cell and Molecular Biology, 24, 210-217.
2.
Callahan, L.A., & Supinski, G.S. (2005). Downregulation of diaphragm electron transport chain and glycolytic enzyme gene expression in sepsis. Journal of Applied Physiology, 99, 1120-1126.
3.
Chae, I.H., Park, K.W., Kim, H.S., & Oh, B.H. (2004). Nitric oxide-induced apoptosis is mediated by Bax/Bcl-2 gene expression, transition of cytochrome C, and activation of caspase-3 in rat vascular smooth muscle cells. Clinica Chimica Acta , 341, 83-91.
4.
Chao, C., Saito, S., Kang, J., Anderson, C.W., Appella, E., & Xu, Y. (2000). p53 transcriptional activity is essential for p53-dependent apoptosis following DNA damage. EMBO Journal , 19, 4967-4975.
5.
Childs, A.C., Phaneuf, S.L., Dirks, A.J., Phillips, T., & Leeuwenburgh, C. (2002). Doxorubicin treatment in vivo causes cytochrome C release and cardiomyocyte apoptosis, as well as increased mitochondrial efficiency, superoxide dismutase activity, and Bcl-2:Bax ratio. Cancer Research, 62, 4592-4598.
6.
Didion, S.P., & Faraci, F.M. (2005). Ceramide-induced impairment of endothelial function is prevented by CuZn superoxide dismutase overexpression. Arteriosclerosis, Thrombosis, and Vascular Biology, 25, 90-95.
Falk, D.J., Deruisseau, K.C., Van Gammeren, D.L., Deering, M.A., Kavazis, A.N., & Powers, S.K. (2006). Mechanical ventilation promotes redox status alterations in the diaphragm. Journal of Applied Physiology, 101, 1017-1024.
9.
Fujimura, N., Sumita, S., Aimono, M., Masuda, Y., Shichinohe, Y., Narimatsu, E., et al. (2000). Effect of free radical scavengers on diaphragmatic contractility in septic peritonitis. American Journal of Respiratory and Critical Care Medicine, 162, 2159-2165.
10.
Gosker, H.R., Wouters, E.F., van der Vusse, G.J., & Schols, A.M. (2000). Skeletal muscle dysfunction in chronic obstructive pulmonary disease and chronic heart failure: Underlying mechanisms and therapy perspectives. American Journal of Clinical Nutrition, 71, 1033-1047.
11.
Gutteridge, J.M., & Mitchell, J. (1999). Redox imbalance in the critically ill. British Medical Bulletin, 55, 49-75.
12.
Ihrie, R.A., Reczek, E., Horner, J.S., Khachatrian, L., Sage, J., Jacks, T., et al. (2003). PERP is a mediator of p53-dependent apoptosis in diverse cell types. Current Biology, 13, 1985-1990.
13.
Itoh, M., Oh-Ishi, S., Hatao, H., Leeuwenburgh, C., Selman, C., Ohno, H., et al. (2004). Effects of dietary calcium restriction and acute exercise on the antioxidant enzyme system and oxidative stress in rat diaphragm. American Journal of Physiology: Regulatory, Integrative and Comparative Physiology, 287, R33-R38.
14.
Jaffer, S.O.L., Sunkara, S., Sabo, E., & Burstein, D.E. (2007). Immunohistochemical detection of antiapoptotic protein X-linked inhibitor of apoptosis in mammary carcinoma . Human Pathology, 38, 864-870.
15.
Kleinberg, L., Lie, A.K., Florenes, V.A., Nesland, J.M., & Davidson, B. (2007). Expression of inhibitor-of-apoptosis protein family members in malignant mesotheliomas. Human Pathology, 38, 986-994.
16.
Kolbeck, R.C., She, Z.W., Callahan, L.A., & Nosek, T.M. (1997). Increased superoxide production during fatigue in the perfused rat diaphragm. American Journal of Respiratory and Critical Care Medicine, 156, 140-145.
17.
Laghi, F., Cattapan, S.E., Jubran, A., Parthasarathy, S., Warshawsky, P., Choi, Y.S., et al. (2003). Is weaning failure caused by low-frequency fatigue of the diaphragm? American Journal of Respiratory and Critical Care Medicine, 167, 120-127.
18.
Li, X., Moody, M.R., Engel, D., Walker, S., Clubb, F.J., Jr., Sivasubramanian, N., et al. (2000). Cardiac-specific overexpression of tumor necrosis factor-alpha causes oxidative stress and contractile dysfunction in mouse diaphragm. Circulation, 102, 1690-1696.
19.
Mayr, M., Hu, Y., Hainaut, H., & Xu, Q. (2002). Mechanical stress-induced DNA damage and rac-p38MAPK signal pathways mediate p53-dependent apoptosis in vascular smooth muscle cells. Journal of the Federation of American Societies for Experimental Biology, 16, 1423-1425.
20.
Nethery, D., Callahan, L.A., Stofan, D., Mattera, R., DiMarco, A., & Supinski, G. (2000). PLA(2) dependence of diaphragm mitochondrial formation of reactive oxygen species. Journal of Applied Physiology , 89, 72-80.
21.
Nowak, M., Koster, C., & Hammerschmidt, M. (2005). PERP is required for tissue-specific cell survival during zebrafish development. Cell Death and Differentiation, 12, 52-64.
22.
Pelech, S., Sutter, C., & Zhang, H. (2003). Kinetworks protein kinase multiblot analysis. Methods in Molecular Biology, 218, 99-111.
23.
Pierce, J.D., Clancy, R.L., Smith-Blair, N., & Kraft, R. (2002). Treatment and prevention of diaphragm fatigue using low-dose dopamine. Biological Research for Nursing, 3, 140-149.
24.
Pierce, J.D., Goodyear-Bruch, C., Hall, S., & Clancy, R.L. (2006). Effect of dopamine on rat diaphragm apoptosis and muscle performance. Experimental Physiology, 91, 731-740.
25.
Pierce, J.D., Goodyear-Bruch, C., Jegathesan, J., & Clancy, R. (2004). Diaphragm apoptosis: Alleviation by dopamine. Critical Care Medicine, 32(Suppl), A121.
26.
Pierce, J.D., Jegathesan, J., Arnett, M.D., Reed, G.D., Masure, R.D., & Clancy, R.D. (2004). Dopamine alleviation of reactive oxygen species mediated diaphragm fatigue. Journal of the Federation of American Societies for Experimental Biology, 18, Abstract no. 702.702. 2004 Experimental Biology meeting abstract.
27.
Powers, S.K., Kavazis, A.N., & DeRuisseau, K.C. (2005). Mechanisms of disuse muscle atrophy: Role of oxidative stress. American Journal of Physiology: Regulatory , Integrative and Comparative Physiology, 288, R337-344.
28.
Qin, Y.M., Auh, S., Blokh, L., Long, C., Gagnon, I., & Hamann, K.J. (2007). TNF-alpha induces transient resistance to Fas-induced apoptosis in eosinophilic acute myeloid leukemia cells. Cellular and Molecular Immunology, 4, 43-52.
29.
Reczek, E.E., Flores, E.R., Tsay, A.S., Attardi, L.D., & Jacks, T. (2003). Multiple response elements and differential p53 binding control PERP expression during apoptosis. Molecular Cancer Research, 1, 1048-1057.
30.
Sakudo, A., Lee, D.C., Saeki, K., Nakamura, Y., Inoue, K., Matsumoto, Y., et al. (2003). Impairment of superoxide dismutase activation by N-terminally truncated prion protein (PrP) in PrP-deficient neuronal cell line. Biochemical and Biophysical Research Communications , 308, 660-667.
31.
Simon, H.U., Haj-Yehia, A., & Levi-Schaffer, F. (2000). Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis, 5, 415-418.
32.
Simpson, J.A., Van Eyk, J., & Iscoe, S. (2004). Respiratory muscle injury, fatigue and serum skeletal troponin I in rat. Journal of Physiology, 554(Part 3), 891-903.
33.
Siu, P.M., Pistilli, E.E., Butler, D.C., & Alway, S.E. (2005). Aging influences cellular and molecular responses of apoptosis to skeletal muscle unloading. American Journal of Physiology: Cell Physiology, 288, C338-C349.
34.
Smith-Blair, N.J., Pierce, J.D., & Clancy, R.L. (2003). The effect of dobutamine infusion on fractional diaphragm thickening and diaphragm blood flow during fatigue. Heart and Lung, 32, 111-120.
35.
Stangel, M., Zettl, U.K., Mix, E., Zielasek, J., Toyka, K.V., Hartung, H.P., et al. (1996). H2O2 and nitric oxide-mediated oxidative stress induce apoptosis in rat skeletal muscle myoblasts. Journal of Neuropathology and Experimental Neurology, 55, 36-43.
36.
Supinski, G., Stofan, D., Ciufo, R., & DiMarco, A. (1997). N-acetylcysteine administration alters the response to inspiratory loading in oxygen-supplemented rats. Journal of Applied Physiology, 82, 1119-1125.
37.
Tews, D.S., Behrhof, W., & Schindler, S. (2005). Expression patterns of initiator and effector caspases in denervated human skeletal muscle. Muscle and Nerve , 31, 175-181.
38.
Vassilakopoulos, T., & Petrof, B.J. (2004). Ventilator-induced diaphragmatic dysfunction. American Journal of Respiratory and Critical Care Medicine, 169, 336-341.
39.
Wei, Y.H., & Lee, H.C. (2002). Oxidative stress, mitochondrial DNA mutation, and impairment of antioxidant enzymes in aging. Experimental Biology and Medicine (Maywood), 227, 671-682.
40.
Yen, G.C., & Hsieh, C.L. (1997). Antioxidant effects of dopamine and related compounds . Bioscience, Biotechnology, and Biochemistry, 61, 1646-1649.
41.
Yokoyama, I., Negita, M., Liu, D.G., Nagasaka, T., Kobayashi, T., Hayakawa, A., et al. (2002). Prevention of free-radical induced apoptosis by induction of human recombinant Cu, Zn-SOD in pig endothelial cells. Transplant International, 15, 220-225.
42.
Yu, Y., Wang, J., Yuan, H., Qin, F., Zhang, N., Li, Y.Y., et al. (2006). Characterization of human dopamine responsive protein DRG-1 that binds to p75NTR-associated cell death executor NADE. Brain Research, 1100, 13-20.
43.
Zhao, L.M., Xiong, S.D., Niu, R.J., Xu, Y.J., & Zhang, Z.X. (2003). Effect of Shen-Mai injection and aminophylline on diaphragmatic muscle cell apoptosis and related gene expression in rats with chronic hypoxia. Zhonghua Jie He He Hu Xi Za Zhi, 26, 606-609.
