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Effects of clearance of superoxide anion by catechin on the expression of NO and eNOS and apoptosis in endothelial progenitor cells induced by angiotensin II
WU Li-Yuan, DANG Xi-Qiang, HE Xiao-Jie, YI Zhu-Wen
Chinese Journal of Contemporary Pediatrics ›› 2009, Vol. 11 ›› Issue (06) : 476-480.
PDF(1160 KB)
PDF(1160 KB)
Effects of clearance of superoxide anion by catechin on the expression of NO and eNOS and apoptosis in endothelial progenitor cells induced by angiotensin II
OBJECTIVE: To evaluate the effect of clearance of superoxide anion (O2—?) by catechin on the expression of nitrogen monoxidum (NO) and endothelial nitricoxide synthase (eNOS) and apoptosis in endothelial progenitor cells (EPCs) induced by angiotensin II (Ang II). METHODS: The marrow endothelial progenitor cells of Sprague-Dawley rats were isolated and assigned to control (no treatment), Ang II treatment and Ang II + catechin treatment groups. After 48 hrs of culture, the concentration of O2—?in the supernate was measured by the NBT method, and NO concentration in the supernate was measured by the nitrate reductase method; the apoptosis rate of EPCs was detected by the TUNEL method; the mRNA expression of eNOS was detected by RT-PCR; the protein expression of eNOS was detected by Western blot analysis. RESULTS: Ang II of 10-6 mol/L was determined as the suitable concentration for cell induction by the MTT test. Catechin of 400 mg/L was determined as an advisable intervention dosage. The apoptosis rate of EPCs in the control, the Ang II and the Ang II+catechin treatment groups were 24.8±1.2‰, 541.8±7.7‰ and 168.7±3.5‰, respectively, and there were significant differences among the three groups (P<0.01). The O2—? concentration in the Ang II and the Ang II+catechin treatment groups (81.7±3. 6 and 62.3±2. 2 U/L respectively) was significantly higher than that in the control group (33.7±2.8 U/L) (P<0.01). An increased NO concentration was also found in the Ang II (189. 8±9.0 μmol/L) and the Ang II+catechin treatment groups (276.4±10.1 μmol/L) compared with that in the control group (105.8±9.8 μmol/L) (P<0.01). There were significant differences in the concentrations of O2—? and NO between the Ang II and the Ang II+catechin treatment groups (P<0.05). The mRNA (P<0.05) and protein expression (P<0.01) of eNOS in the Ang II and the Ang II+catechin treatment groups increased significantly compared with those in the control group. The Ang II+catechin treatment group showed increased eNOS protein expression compared with the Ang II group (P<0.05). CONCLUSIONS: Ang II may induce the generation of O2—?, inactivate NO and increase gene and protein expression of eNOS in EPCs. Catechin might decrease the apoptosis of EPCs through the effective clearance of O2—? and the reduction of NO inactivation and of eNOS protein uncoupling.［Chin J Contemp Pediatr, 2009, 11 (6):476-480］
Catechin / Angiotensin II / Endothelial progenitor cell / Superoxide anion / Nitrogen monoxidum / Rats
［1］Asahara T, Murohara T, Sullivan A, Silver M, van der Zee R, Li T, et al. Isolation of putative progenibr endothelial cells for angiogenesis［J］.Science, 1997, 275(5302):964-967.
［2］Ingram DA, Mead LE, Moore DB, Woodard W, Fenoglio A, Yoder MC. Vessel wall-derived endothelial cells rapidly proliferate because they contain a complete hierarchy of endothelial progenitor cells［J］.Blood, 2005, 105(7):2783-2786.
［3］Urbich C, Dmmeler S. Endothelial progenitor cells Characterization and role in vascular biology［J］.Circ Res, 2004, 95(4):343-353.
［4］Papaharalambus CA, Griendling KK. Basic mechanisms of oxidative stress and reactive oxygen species in cardiovascular injury［J］.Trends Cardiovasc Med, 2007, 17(2):48-54.
［5］Thum T, Fraccarollo D, Schultheiss M, Froese S, Galuppo P, Widder JD, et al. Endothelial nitric oxide synthase uncoupling impairs endothelial progenitor cell mobilization and function in diabetes［J］. Diabetes, 2007, 56(3):666-674.
［6］何小解，许自川，郑洪，党西强，易著文，曾雪琪.大鼠骨髓内皮祖细胞分离与体外培养条件研究［J］.世界科技研究与发展，2008，30（2）：198-201.
［7］何小解，易著文，党西强，吴小川, 何庆南, 卢向阳,等.儿茶素对肾病综合征大鼠血清氧自由基与抗氧化酶的影响［J］.肾脏病与透析肾移植杂志, 2002, 11( 4):342-346.
［8］林红英，沈子龙，张奕华，计晓斌，王伟强，彭司勋. NO调控剂（S，S）-ZX-5及其对映体对血管内皮细胞NO释放及eNOS酶活性影响研究［J］.中国药科大学学报，2006, 37(6):531-534.
［9］马吉祥,苏军英,李会庆,王显军, 颜云.大豆异黄酮诱导胃癌细胞凋亡作用研究［J］. 中国公共卫生, 2003, 19(4):434-436.
［10］吴杰，陈香美.血管病变在1005例IgA肾病中的意义及其影响因素［J］.中华医学杂志, 2003, 83(4):289-293.
［11］Zhang C, Yang J, Jennings LK. Leukocyte-derived myeloperoxidase amplifies high-glucose-induced endothelial dysfunction through interaction with high-glucose-stimulated, vascular non-leukocyte-derived reactive oxygen species［J］. Diabetes, 2004, 53（11）:2950-2959.
［12］Griendling KK, Sorescu D, Ushio-Fukai M. NAD（P）H oxidase:role in cardiovascular biology and diseaee［J］.Circ Res, 2000, 86（5）：494-501.
［13］Brosnan MJ, Hamilton CA, Graham D, Lygate CA, Jardine E, Dominiczak AF. Irbesartan lowers superoxide levels and increases nitric oxide bioavailability in blood vessels from spontaneously hypertensive stroke-prone rat［J］.J Hypertens, 2002, 20(2)：281-286.
［14］Manishit, Hano T, Nishio I. Estrogen reduces angiotensin-Ⅱ induced acceleration of senesce1nce in endothelial progenitor cells［J］. Hypertens Rev, 2005, 28(3):263-271.
［15］Dernbach E, Urbich C, Brandes RP, Hofmann WK, Zeiher AM, Dimmeler S. Antioxidative stress-associated genes in circulating progenitor cells: Evidence for enhanced resistance againstoxidative stress［J］.Blood, 2004, 104(12):3591-3597.
［16］Sarr M, Chataigneau M, Martins S, Schott C, El Bedoui J, Oak MH，et al. Red Wine polyphenols prevent angiotensinⅡ-induced hypertension and endothelial in rats：role of NADPH oxidase［J］. Cardivasc Res, 2006, 71(4)：794-802.
［17］Maeda K, Kuzuya M, Cheng XW, Asai T, Kanda S, TamayaMori N，et al. Green tea polyphenol inhibit the cultured smooth Muscles cell Invasion through the basement barrier［J］. Atherosolerosis, 2003, 166(1):23-30.
［18］何小解，卢向阳，刘永乐，易著文，田云.儿茶素对肾病大鼠一氧化氮和蛋白尿生成的影响［J］.食品科学，2003，24（1）：121-124.
