论文部分内容阅读
目的 :探讨连豆清脉方对ANGⅡ诱导内皮细胞凋亡模型AT1R、AT2R、caspase8和caspase9 mRNA表达的影响。方法 :体外培养大鼠动脉内皮细胞株(RAOEC),不同浓度梯度连豆清脉方煎剂干预ANGⅡ诱导RAOEC凋亡,采用流式细胞仪检测ANGⅡ诱导凋亡改变,RT-PCR荧光相对定量检测AT1R、AT2R、caspase8和caspase9 mRNA水平变化,组间差异统计性分析采用Students T-test。结果 :ANGⅡ诱导模型组内皮细胞24 h凋亡率为(30.4±5.2)%,连豆清脉方(1000μg)干预组内皮细胞24 h凋亡率为(11.2±3.4)%,连豆清脉方能够部分抑制ANGⅡ诱导内皮细胞作用,差异具有统计学意义(P<0.05)。RT-PCR荧光相对定量显示ANGⅡ组AT1R、AT2R、caspase8和caspase9 mRNA水平显著升高(P<0.05),连豆清脉方(1000μg)干预组AT1R和AT2R mRNA表达水平上调,caspase9 mRNA水平显著下调(P<0.05)。结论 :连豆清脉方脉方能够干预caspase 9 mRNA表达,影响caspase 9活化所导致的下游凋亡级联反应,抑制ANGⅡ诱导的内皮细胞凋亡。
Objective: To investigate the effect of Liantou Qingmai Decoction on expression of AT1R, AT2R, caspase8 and caspase9 mRNA in endothelial cell apoptosis induced by angiotensin Ⅱ. Methods: Rat RAOEC was cultured in vitro. Angiotensin II (Angiotensin II) -mediated apoptosis was induced by different concentrations of Lvhuoqingmaifang decoction. Angiotensin II (AngⅡ) -mediated apoptosis was detected by flow cytometry. Relative quantification AT1R, AT2R, caspase8 and caspase9 mRNA levels, statistical analysis of differences between groups using Students T-test. RESULTS: The apoptotic rate of endothelial cells in ANGⅡ-induced group was (30.4 ± 5.2)% at 24 h, that of LIAOHUANGMAI (1000μg) at 24 h was (11.2 ± 3.4)%, Could partly inhibit the effect of ANGⅡ on endothelial cells, the difference was statistically significant (P <0.05). Relative quantification of RT-PCR showed that the mRNA levels of AT1R, AT2R, caspase8 and caspase9 in ANGⅡ group were significantly increased (P <0.05), while the expression of AT1R and AT2R mRNA was up-regulated and the level of caspase9 mRNA in ANGⅡ group was significantly decreased (P <0.05). Conclusion: Liantou recipe can interfere with the expression of caspase 9 mRNA, affect the downstream cascade of caspase 9 activation and inhibit the apoptosis of endothelial cells induced by angiotensin Ⅱ.