论文部分内容阅读
目的探讨去天门冬氨酸血管紧张素Ⅰ(DAA-Ⅰ)对大鼠心肌微血管内皮细胞(CMECs)缺血/再灌注损伤的影响。方法分离培养大鼠CMECs,建立模拟缺血/再灌注模型,完全随机分组,分为对照组、模拟缺血/再灌注组(SI/R组)、模拟缺血/再灌注+DAA-Ⅰ(2.5、3.75、5.0μmol/L)组。MTT检测细胞增殖能力,细胞划痕实验检测细胞迁移能力,TUNEL法检测细胞凋亡,采用Western blot检测GRP78/Bip、CHOP/GADD153、pro-caspase-12、cleaved-caspase-12蛋白的表达。结果与对照组相比较,SI/R组CMECs增殖能力明显降低(P<0.01),凋亡率显著上升(24.85%±0.67%比3.55%±0.11%,P<0.01)。与SI/R组相比,SI/R+DAA-Ⅰ组细胞增殖能力明显升高(P<0.01)并呈剂量依赖性,细胞迁移率上升(P<0.01)并呈剂量依赖性,凋亡率明显下降(15.18%±0.40%比24.85%±0.67%,P<0.01)并呈剂量依赖性。与对照组相比较,SI/R组和SI/R+DAA-Ⅰ组的GRP78、CHOP/GADD153、cleaved-caspase-12表达明显上升(均为P<0.01)。与SI/R组相比,SI/R+DAA-Ⅰ组的GRP78、CHOP/GADD153、cleaved-caspase-12表达明显降低(均为P<0.01)。结论 DAA-Ⅰ可显著抑制缺血/再灌注损伤诱导的CMECs凋亡,促进CMECs存活,改善细胞功能,其保护作用可能与下调ERS相关蛋白的表达有关。
Objective To investigate the effects of aspartate angiotensin Ⅰ (DAA-Ⅰ) on ischemia / reperfusion injury in rat cardiac microvascular endothelial cells (CMECs). Methods CMECs were isolated and cultured in vitro. The models of simulated ischemia / reperfusion were established and randomly divided into control group, SI / R group, simulated ischemia / reperfusion + DAA-Ⅰ 2.5, 3.75, 5.0 μmol / L). Cell proliferation was assessed by MTT. Cell migration was assessed by cell scratch assay. Apoptosis was detected by TUNEL assay. The expressions of GRP78 / Bip, CHOP / GADD153, pro-caspase-12 and cleaved-caspase-12 were detected by Western blot. Results Compared with the control group, the proliferation of CMECs in SI / R group was significantly decreased (P <0.01), and the apoptosis rate was significantly increased (24.85% ± 0.67% vs 3.55% ± 0.11%, P <0.01). Compared with SI / R group, the proliferation of SI / R + DAA-Ⅰ group was significantly increased (P <0.01) and in a dose-dependent manner, the cell migration rate increased (P <0.01) The rate was significantly decreased (15.18% ± 0.40% vs 24.85% ± 0.67%, P <0.01) in a dose-dependent manner. Compared with the control group, the expressions of GRP78, CHOP / GADD153 and cleaved-caspase-12 in SI / R group and SI / R + DAA-Ⅰ group were significantly increased (all P <0.01). Compared with SI / R group, the expressions of GRP78, CHOP / GADD153 and cleaved-caspase-12 in SI / R + DAA-Ⅰ group were significantly decreased (all P <0.01). Conclusion DAA-Ⅰ can significantly inhibit the apoptosis of CMECs induced by ischemia / reperfusion injury and promote the survival of CMECs and improve the cell function. The protective effect of DAA-Ⅰ may be related to the down-regulation of ERS-related protein expression.