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目的观察阿托伐他汀钙对血管紧张素Ⅱ的促人肠系膜动脉平滑肌细胞增殖和其诱导的平滑肌细胞内NADPH氧化酶-活性氧通路的干预作用。方法用组织贴块法培养的第2～4代人肠系膜动脉平滑肌细胞作为实验标本;以四甲基偶氮唑盐(MTF)法比色测定各组平滑肌细胞增殖率;流式细胞仪检测被双氢乙酰乙酸-二氯荧光黄(DCFH-DA)标记的细胞内活性氧;反转录聚合酶链式反应(RT-PCR)方法检测细胞内p22phox蛋白mRNA的表达变化。结果 (1)与空白对照组比较,加入血管紧张素Ⅱ药物组的细胞增殖率、细胞内活性氧物质生成量、细胞内p22phox mRNA的表达均明显增高(P<0.01)。(2)与浓度为10~(-6)mol/L血管紧张素Ⅱ孵育组比较,血管紧张素Ⅱ和阿托伐他汀钙共孵育组的细胞增殖率[阿托伐他汀组(0.242±0.018)比血管紧张素Ⅱ组(0.359±0.024),P<0.01]、细胞内活性氧[阿托伐他汀组(130±29)比血管紧张素Ⅱ组(180±34),P<0.01]、细胞内p22phox蛋白mRNA的表达[阿托伐他汀组(0.47±0.07)比血管紧张素Ⅱ组(0.71±0.05),P<0.01]均降低。结论阿托伐他汀钙可以部分抑制血管紧张素Ⅱ的促人肠系膜动脉平滑肌细胞增殖和其诱导的细胞内NADPH氧化酶-活性氧通路的作用。
Objective To observe the effect of atorvastatin calcium on the proliferation of human mesenteric artery smooth muscle cells induced by angiotensin Ⅱ and the intervention of NADPH oxidase - reactive oxygen species pathway in smooth muscle cells. Methods The second to fourth generation of human mesenteric artery smooth muscle cells cultured in tissue block method were used as experimental specimens. The proliferation of smooth muscle cells in each group was determined by MTT colorimetric assay. Flow cytometry The intracellular reactive oxygen species labeled with DCFH-DA and the expression of p22phox mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR). Results (1) Compared with the blank control group, the cell proliferation rate, intracellular reactive oxygen species production and the expression of intracellular p22phox mRNA in the group receiving angiotensin Ⅱ were significantly increased (P <0.01). (2) Compared with the group incubated with angiotensin Ⅱ at a concentration of 10 -6 mol / L, the cell proliferation rate of the group incubated with angiotensin Ⅱ and atorvastatin calcium [atorvastatin group (0.242 ± 0.018 ) Was significantly higher than that of the angiotensin Ⅱ group (0.359 ± 0.024, P <0.01). The intracellular reactive oxygen species in the atorvastatin group (130 ± 29) and the angiotensin Ⅱ group (180 ± 34) The expression of intracellular p22phox protein mRNA in the atorvastatin group (0.47 ± 0.07) was significantly lower than that in the angiotensin Ⅱ group (0.71 ± 0.05), P <0.01. Conclusions Atorvastatin calcium can partially inhibit the proliferation of human mesenteric artery smooth muscle cells induced by angiotensin Ⅱ and the effect of its induction of intracellular NADPH oxidase - reactive oxygen species pathway.