论文部分内容阅读
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annexin Ⅴ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340 nm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 effects of Astragaloside Ⅳ on the proliferation of abnormal vascular smooth muscle cells (VSMCs). Methods VSMCs obtained from thoracic aorta of SD rats were cultured in vitro. The changes of cell proliferation, cell apoptosis, NO and Ca concentration before and after Astragaloside Ⅳ administration in VSMCs were measured. Cell proliferation was measured with tetrazolium salt WST-1 assay. Cell apoptosis was assayed with flow cytometry through the detection of annexin Ⅴ. NO and in-tracellular Ca concentration were measured by using confocal laser scanning microscopy with diaminofluorescein diacetate and Flue-3/AM, respectively. Cell aldose reductase (AR) activity was measured with absorbent spectrum of NADPH at 340 nm. Results WST assay showed that cell proliferation induced by serum was significantly inhibited by Astragaloside Ⅳ(P<0.01 ). Analysis on flow cytometry revealed that Astragaloside Ⅳ could enhance the apoptotic rate of VSMCs (P<0.01). The AR activity of VSMCs was markedly inhibited by both Astragaloside Ⅳ and an AR inhibitor, epalrestat (P<0.01). NO production was significantly enhanced after the administration of Astragaloside Ⅳ or interleukin-1β. Furthermore, Astragaloside Ⅳ increased cytosolic Ca significantly. Conclusion Astragaloside Ⅳ may inhibit the proliferation of VSMCs associated with serum by inhibiting AR activity, augmenting apoptosis, and increasing the levels of cytosolic Ca and NO production in VSMCs.