富含微量元素矿泉水对人血管平滑肌细胞生长增殖的影响

摘要

Objective To study the influence of mineral water rich in microelement on proliferation of human vascular smooth muscle cells(HVSMCs) in vitro. Method Ten samples of human great saphenous vein of patients undergoing coronary artery bypass surgery were obtained. Two kinds of DMEM media including Jinchuan mineral water (MW-DMEM)and double distilled water(DDW-DMEM) were observed. The free tissue formation time and the "peak-valley" time of HVSMCs cultured in these 2 kinds of DMEM media were observed and recorded.A cell proliferating model of HVSMCs was induced by angiotensin Ⅱ. 3-(4,5-dimethylthiazol-2-yl)-2,5-dipheny-ltetrazolium bromide (MTT) was used to determine the cells growth. The cell was confirmed to be HVSMCs by im-munohistochemistry and electromicroscopy analysis. Results The HVSMCs in MW-DMEM group emigrating from the tissue explanted within (3.7±0.2) days while the HVSMCs in DDW-DMEM group explanted within (8.5±0.6)days. The time of HVSMCs forming a 'peak-trough state' from a adherence state in MW-DMEM group was (8.3±0.2)days and it was (7.1±0.5)days in DDW-DMEM group. The result showed that H VSMCs proliferation induced by Ang Ⅱ could be inhibited by Jinchuan mineral water. HVSMCs were confirmed under electron microscope of the integrity of cell morphology and the surface is rich in microvilli. HVSMCs cytoplasm was rich in myofilaments and was in a longitudinal and parallel arrangement in intracellular dense areas Anti-α-actin antibodies with immuno-histochemical staining showed lots of stained buffy bundles in cell cytoplasm. Conclusions The results show that Jinchuan mineral water rich in microelement and mineral can relieve injury of HVSMCs. Meanwhile HVSMCs prolif-eration induced by Ang Ⅱ can be inhibited by Jinchuan mineral water.%目的 研究富含微量元素矿泉水对体外培养血管平滑肌细胞(HVSMCs)生长增殖的影响.方法 无菌取12例冠状动脉旁路移植术患者术中废弃的大隐静脉,应用金山矿泉水配制的DMEM培养液(MW-DMEM)和双蒸水配制的DMEM培养液(DDW-DMEM)分别培养平滑肌细胞,分为金川矿泉水组和双蒸水组.观察记录2种培养液培养的HVSMCs游出组织块的时间及细胞呈"峰"-"谷"样时间;建立血管紧张素Ⅱ诱导HVSMCs增殖模型,采用MTT比色法观察2种培养液对Ang Ⅱ诱导HVSMCs增殖抑制作用;应用电镜和免疫组织化学染色法观察鉴定HVSMCs.结果 金川矿泉水组培养的HVSMCs贴壁时间为(3.7±0.2)d,双蒸水组为(8.5±0.6)d;金川矿泉水组培养的HVSMCs自贴壁后到长成"峰"-"谷"样形态的时间为(8.3±0.2)d,双蒸水组为(7.1±0.5)d;MTT比色法测定金川矿泉水组培养的HVSMCs的增殖活度明显降低;电镜观察MW-DMEM培养的HVSMCs细胞形态完整,表面有丰富的微绒毛.HVSMCs胞浆内富含肌丝,纵向平行排列,胞内有密体;抗-actin平滑肌肌动蛋白免疫组织化学染色后可见胞浆内染成褐色呈细丝网状.结论 富含微量元素和矿物质的金川矿泉水具有修复血管平滑肌细胞损伤和抑制血管紧张素Ⅱ诱导HVSMCs增殖的作用.