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目的探讨从黄芪提取物黄芪甲苷Ⅳ对异常血管平滑肌细胞增殖的抑制作用。方法离体培养大鼠主动脉平滑肌细胞,分别测定黄芪甲苷Ⅳ作用前后异常血管平滑肌细胞的增殖、凋亡、细胞一氧化氮(NO)和钙离子浓度的变化。用WST法测定细胞的增殖;细胞凋亡通过annexinV标记法用流式细胞仪测定;细胞内NO和钙离子用荧光素DAF-2和Flue-3/AM分别标记,运用激光共聚焦显微镜测定其浓度变化;细胞醛糖还原酶的活性通过测定NADPH在340nm的吸收光谱来表征。结果黄芪甲苷Ⅳ显著抑制了由血清诱导的平滑肌细胞增殖(P<0.01);流式细胞测定表明黄芪甲苷Ⅳ促进平滑肌细胞的凋亡(P<0.01);黄芪甲苷Ⅳ和醛糖还原酶抑制剂依帕斯他显著抑制了醛糖还原酶的活性(P<0.01);在黄芪甲苷Ⅳ作用下,平滑肌细胞NO和钙离子浓度增加。结论黄芪甲苷Ⅳ可能通过抑制醛糖还原酶的活性,促进细胞凋亡,增加NO的产生和钙离子浓度的途径来抑制异常血管平滑肌细胞的增殖。
Objective To investigate the inhibitory effect of astragaloside Ⅳ from Astragalus extract on the proliferation of abnormal vascular smooth muscle cells. Methods Rat aortic smooth muscle cells were cultured in vitro and the changes of proliferation, apoptosis, nitric oxide (NO) and calcium concentration in vascular smooth muscle cells were measured before and after astragaloside IV treatment. Cell proliferation was determined by WST method. Apoptosis was determined by annexin V labeling method. Intracellular NO and Ca2 + were labeled with fluorescein DAF-2 and Flue-3 / AM, respectively. Concentration; the cellular aldose reductase activity was characterized by measuring the absorption spectrum of NADPH at 340 nm. Results Astragaloside IV significantly inhibited the proliferation of smooth muscle cells induced by serum (P <0.01). Flow cytometry analysis showed that astragaloside IV promoted the apoptosis of smooth muscle cells (P <0.01); Astragaloside Ⅳ and aldose reductase Enzyme inhibitor Enzyme significantly inhibited the activity of aldose reductase (P <0.01); in Astragaloside IV, the concentration of NO and calcium in smooth muscle cells increased. Conclusion Astragaloside Ⅳ may inhibit the proliferation of abnormal vascular smooth muscle cells by inhibiting the activity of aldose reductase, promoting cell apoptosis, increasing the production of NO and the concentration of calcium ion.