益肾散结复方及拆方对阿霉素诱导的HBZY-1肾小球系膜细胞凋亡的影响

Effect of Kidney-reinforcing and Mass-disintegrating Compound and Decomposed Formula on Adriamycin-induced Apoptosis of HBZY-1 Glomerular Mesangial Cells

目的:探讨益肾散结复方及拆方对阿霉素毒性诱导的HBZY-1大鼠肾小球系膜细胞凋亡的影响及可能的调控机制。方法:实验分为以下7组:正常组、阿霉素肾病模型组、盐酸贝那普利对照组、温肾益精化瘀组、滋阴益肾化瘀组、益气化瘀组、益肾散结复方组。将复苏的HBZY-1培养6~8 h待细胞同步化后,阿霉素按照不同浓度加入,筛选最佳建模浓度和接种密度,利用实时无标记细胞分析RTCA系统检测细胞增殖,选择药物处理的最适浓度及时间,培养16 h后采用流式细胞术Annexin V-FITC/PI双染法检测细胞凋亡率。结果:阿霉素能显著诱导HBZY-1细胞凋亡;益肾散结复方组、温肾益精化瘀拆方组在药物处理浓度为1000 mg/L、滋阴益肾化瘀拆方组与益气化瘀拆方组在药物浓度为800 mg/L时,与盐酸贝那普利对照组药物浓度为200 mg/L相比,有较好的抑制细胞凋亡的作用。结论:益肾散结复方及其拆方均可抑制肾小球系膜细胞HBZY-1的晚期凋亡。

Objective:To investigate the effect of Kidney-reinforcing and Mass-disintegrating Compound and its decomposed formula on apoptosis of glomerular mesangial cells induced by Adriamycin in HBZY-1 rats and its possible regulation mechanism.Methods:The experiment was divided into the following 7 groups:normal group,Adriamycin nephropathy model group,benazepril hydrochloride group,warming the kidney to replenish essence group,replenishing Yin to tonify Kidney,remove blood stasis group and Kidney-reinforcing and Mass-disintegrating Compound group;After the resuscitated HBZY-1 was cultured for 6-8h to be synchronized with cells,Adriamycin was added at different concentrations to select the best modeling concentration and seeding density.Using xCELLigence RTCA to detect cell proliferation and select the optimal concentration and time of drug treatment.After 16 h of culture,the cell apoptosis rate was detected by flow cytometry Annexin V-FITC/PI double staining method.Results:Adriamycin significantly induced apoptosis of HBZY-1;When the concentration of Kidney-reinforcing and Mass-disintegrating Compound group and warming the kidney to replenish essence group were 1000 mg/L,the concentration of replenishing Yin to tonify Kidney group and remove blood stasis group were 800 mg/L.they had better inhibition of apoptosis compared with benazepril hydrochloride group.Conclusion:Kidney-reinforcing and Mass-disintegrating Compound and its decomposed formula can inhibit the late apoptosis of glomerular mesangial cells HBZY-1.