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
Department of Pediatrics, Second Xiangya Hospital, Central South University, Changsha 410011, China.
Abstract：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］
WU Li-Yuan,DANG Xi-Qiang,HE Xiao-Jie et al. 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[J]. CJCP, 2009, 11(06): 476-480.
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