补阳还五汤对高盐致HK2细胞凋亡保护作用机制研究

Protective Mechanism of Buyang Huanwu Decoction(补阳还五汤) on HK2 Cell Apoptosis Induced by High Salt

目的 探究补阳还五汤水提物冻干粉对高盐诱导的人肾小管上皮细胞(HK2)损伤的保护作用。方法 采用高盐(150 mmol/L)干预HK2细胞,诱导细胞损伤,通过加入不同剂量的补阳还五汤(0.5%、1%、2%)作用后,分别采用细胞增殖毒性检测法(CCK-8法)检测补阳还五汤对细胞的保护作用,流式细胞术检测细胞凋亡及活性氧表达,ELISA检测细胞培养液中白细胞介素-1β(IL-1β)和白细胞介素-18(IL-18)含量。结果 与空白组相比,高盐组细胞数目明显减少,形态出现异常改变,同时培养液中漂浮细胞明显增多,细胞活性显著降低(P<0.05);细胞凋亡结果显示,高盐组细胞凋亡比例显著提高(P<0.05);ROS检测结果表明,高盐组细胞ROS表达显著增高(P<0.05);ELISA检测结果表明,高盐组细胞上清液中IL-1β、IL-18含量显著降低。与高盐组相比,补阳还五汤可以显著促进细胞增殖,抑制ROS表达及高盐诱导的细胞凋亡反应,减少IL-1β、IL-18等反应(P<0.05),治疗作用呈现一定的剂量依赖性。结论 补阳还五汤能够抑制高盐诱导的HK2细胞凋亡而发挥细胞保护作用。

Objective To explore the protective effect of freeze-dried powder of Buyang Huanwu Decoction(补阳还五汤)on high salt induced injury of human renal tubular epithelial cells(HK2).Method High salt(150 mmol/L)was used to intervene HK2 cells and induce the cell damage.After adding different doses of Buyang Huanwu Decoction(0.5%,1%,2%),the protective effect of Buyang Huanwu Decoction on cells was detected by cell proliferation toxicity test(CCK-8 method),cell apoptosis and reactive oxygen species expression were detected by flow cytometry,and the levels of interleukin 1(IL-1),interleukin 1β(IL-1β)and interleukin 18(IL-18)in cell culture medium were detected by ELISA.Result Compared with that of the blank group,the number of cells in the high salt group was decreased significantly,the morphology changed abnormally,the floating cells in the culture medium increased significantly,and the cell activity decreased significantly(P<0.05).The results of apoptosis showed that the proportion of apoptosis in high salt group was increased significantly(P<0.05).The results of ROS detection showed that the expression of ROS in cells of the high salt group was significantly increased(P<0.05).The results of ELISA showed that the levels of IL-18 and IL-1βin cell supernatant of the high salt group were decreased significantly.Compared with the high salt group,Buyang Huanwu Decoction can significantly promote cell proliferation,inhibit ROS expression and high salt inducing apoptosis,and reduce the levels of IL-1β,IL-18 and other reactions(P<0.05),and the therapeutic effect showed a certain dose dependence.Conclusion Buyang Huanwu Decoction can inhibit HK2 cell apoptosis induced by high salt and plays a cytoprotective role.