Abstract
Objective
To investigate whether the administration of the ultra-filtration extract from Danggui Buxue Decoction (当归补血汤, EDBD) was able to protect cardiomyocytes from oxidative injury of rats induced by hydrogen peroxide (H2O2) and its potential mechanism.
Methods
Myocardial cells from 1- to 2-day-old neonatal rats were cultured in Dulbecco’s modified Eagle’s medium low-glucose and Ham’s F12 medium (1:1), and the cellular injury was induced by H2O2. The ultra-filtration extract mixture from Angelica sinensis and Hedysarum polybotrys was given in three doses of 3.75, 7.5, and 15 mg/mL. Morphological changes of cardiomyocytes were observed by microscope. Survival rate of myocardial cells was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The cardiomyocyte damages were estimated by detecting lactate dehydrogenase (LDH) and creatine kinase (CK) releases in the medium, superoxide dismutase (SOD) activities, and intracellular malondialdehyde (MDA) and myeloperoxidase (MPO) contents. The levels of caspase-3 and heat shock protein 70 (hsp70) mRNA expression in cardiomyocytes were measured by reverse transcription polymerase chain reaction.
Results
The EDBD could protect the cardiomyocytes from H2O2 injury in a dosedependent manner (3.75, 7.50, and 15.00 mg/mL). The EDBD could significantly decrease LDH and CK leakages and intracellular MDA and MPO contents, increase SOD activity, up-regulate hsp70 expression, and down-regulate caspase-3 expression.
Conclusion
The EDBD has protection on cardiomyocytes injured by H2O2 through improving cell antioxidant ability, up-regulating hsp70 expression, and inhibiting caspase-3 activity.
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References
Li GR, Lu CY, Yang SY. Study on Isolation of Angelica Sinensis Dliv. Polysaccharide and its effects on anti-active oxygen free radicals. J Central-south Institute Technol (Chin) 2002;20:747–749.
Deng HJ, Guo YS, Qu YL, Yang XH, Qin L, Wei YM. A study of scavenging action of radix Angelica sinesis recen, wine-Angelica sinesis and oil-Angelica sinesis on oxygen free radical in vivo. Chin Tradit Herbal Drugs (Chin) 2009;40:784–787.
Shang P, Qian AR, Yang TH, Jia M, Mei QB, Cho CH, Zhao WM, Chen ZN. Experimental study of anti-tumor effects of polysaccharides from Angelica sinensis. World J Gastroenterol 2003;9:1963–1967.
Yao BT, ZHAO JX, Wang XX, Lei FF. Experimental study on anti-tumor activity of total hedysarum polybotrys saccharide in vivo. China Tradit Chin Med Pharm (Chin) 2008;23:627–629.
Li YZ, Huang J, Guo HC, Ren B. Chemical constituents from Hedysarum polybotrys and their antitumor activities. Chin Tradit Herbal Drugs (Chin) 2009;40:1195–1198.
Sun YL, Tang J, Gu XH, Li DY. Water-soluble polysaccharides from Angelica sinensis (Oliv.) Diels: preparation, characterization and bioactivity. Int J Biol Macromol 2005;36:283–289.
Guo LW. Significant demand and key problems of membrane science and technology applying to pharmaceutical industry of Chinese material medica. Chin Tradit Herbal Drugs (Chin) 2009;40:1849–1855.
Cao YT, Guo LW, Shi DL, Pan LM, Zhu HX. Advances in studies on ceramic membrane applied in Chinese material medica refining. Chin Tradit Herbal Drugs (Chin) 2010;41:314–317.
Dong Y, Gao RC, Pan Q, Yan XN. Ultrafiltration and nanofiltration membrane separation technology in extracting and purifying oligosaccharides of Rehmannia glutinosa. Chin Tradit Herbal Drugs (Chin) 2008;39:359–363.
Chen SH. A study of using UF in refining Chinese traditional medicine. Water Purific Technol (Chin) 2004;23(2):16–18.
Li YD, ed. An experimental study on angiogenesis promoting effect of the ultra-filtration extract from Angelica Sinensis, Hedysarum Polybotrys and their mixture in ischemic myocardium. Nanjing: Nanjing University of Chinese Medicine; 2005:9–10.
Simpson P, Savion S. Differentiation of rat myocytes in single cell cultures with and without proliferating nonmyocardialcell. Circ Res 1982;50:101–116.
Xiao WM, Jiang BM, Shi YZ, Liu MD, Tang DL, Xin K, et al. Heat shock proteins protect cardiomyocytes against apoptosis induced by hydrogen peroxide through interference with two signal transduction pathways. Chin J Arterioscl (Chin) 2003;11:283–286.
Zhang HB, Wan YT, Wang LZ, Liao CL, Yang HB, Chen LY. Protection and regulation of Hsp70 in cellular stress responses. China Pract Med 2009;4:228–229.
Latchman DS. Heat shock proteins and cardiac protection. Cardiovasc Res 2001; 51: 637–646.
Jiang B, Xiao W, Shi Y, Liu MD, Xiao XZ. Heat shock pretreatment inhibited the release of Smac/DIABLO from mitochondria and apoptosis induced by hydrogen peroxide in cardiomyocytes and C2C12 myogenic cells. Cell Stress Chaper 2005;10:252–262.
Faubel S, Edelstein CL. Caspases as drug targets in ischemic organ injury. Curr Drug Targets Immune Endocr Metabol Disord 2005;5:269–287.
Shen J, Lee W, Gu Y, Tong Y, feng PC, Tong L. Ginkgo biloba extract (EGb761) inhibits mitochondria-dependent caspase pathway and prevents apoptosis in hypoxiareoxygenated cardiomyocytes. Chin Med 2011;23(6):8.
Misra MK, Sarwat M, Bhakuni P, Tuteja R, Tuteja N. Oxidative stress and ischemic myocardial syndromes. Med Sci Monit 2009;15:209–219.
Orrenius S. Reactive oxygen species in mitochondriamediated cell death. Drug Metab Rev 2007;39:443–455.
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Supported by the Key Projects of the National Science and Technology Pillar Program during the Eleventh Five-Year Plan Period (No. 2007BAI37B01) and Major Science and Technology Program of Gansu Province (No. 092NKDA017)
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Li, Yd., Ma, Yh., Zhao, Jx. et al. Protection of ultra-filtration extract from Danggui Buxue Decoction (当归补血汤) on oxidative damage in cardiomyocytes of neonatal rats and its mechanism. Chin. J. Integr. Med. 17, 854–859 (2011). https://doi.org/10.1007/s11655-011-0897-6
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DOI: https://doi.org/10.1007/s11655-011-0897-6