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目的 :探讨宁血益肾颗粒对肾小球系膜细胞凋亡及凋亡相关基因bax、bcl- 2表达的影响 ,以阐明其治疗系膜增生性肾病有效的作用机制 ,为临床利用该药治疗以系膜增生为核心病理环节的多种肾脏疾病提供实验理论依据。方法 :采用肾小球系膜细胞培养方法进行体外研究 ,用不同剂量的宁血益肾颗粒刺激 4～ 6代系膜细胞 16小时 ,MTT法测定系膜细胞增殖指数 ;TUNEL法定量观察凋亡细胞 ;流式细胞术检测凋亡相关基因bax、bcl- 2的表达。结果 :加入宁血益肾颗粒 16小时后 ,系膜细胞出现了典型的凋亡征象 ,诸如细胞内深棕色颗粒形成、核固缩、碎裂等 ;凋亡率显著高于对照组 (P <0 .0 1) ;同时血尿停各组bax、bcl- 2表达亦明显高于对照组 (P <0 .0 1或P <0 .0 5 )。观察其对凋亡、凋亡相关基因的影响 ,为中医治疗以系膜增生为主要病理改变的多种肾脏疾病 (原发性系膜增生性肾病、紫癜性肾炎、IgA肾病、狼疮性肾病等 )提供新的实验依据。结论 :宁血益肾颗粒可能通过调控bax和bcl- 2基因表达诱导系膜细胞凋亡 ,促进肾小球增殖性病变消散。
Objective: To investigate the effect of Ningxue Yishen Granules on the apoptosis of mesangial cells and the expression of apoptosis-related genes bax and bcl-2, so as to elucidate the effective mechanism of mesangial proliferative nephropathy, and to use this drug clinically. It provides experimental theoretical basis for the treatment of various renal diseases with mesangial hyperplasia as the core pathological aspect. METHODS: Mesangial cell culture method was used for in vitro study. Mesangial cells were stimulated with different doses of Ningxue Yishen Granules for 4 to 6 generations for 16 hours. Medullary cell proliferation index was measured by MTT assay; apoptosis was quantified by TUNEL method. The expression of apoptosis-related genes bax and bcl-2 was detected by flow cytometry. RESULTS: After 16 hours of Ningxue Yishen Granules, the mesangial cells showed typical signs of apoptosis, such as the formation of deep brown particles, nuclear condensation, and fragmentation, and the apoptosis rate was significantly higher than that of the control group (P < The expression of bax and bcl-2 was also significantly higher in the hematuria group than in the control group (P <0.01 or P <0.05). Observe its effect on apoptosis-related and apoptosis-related genes, and treat various kidney diseases (main mesangial proliferative nephropathy, purpura nephritis, IgA nephropathy, lupus nephropathy, etc.) in which TCM treats mesangial hyperplasia as the main pathological change. ) Provide new experimental evidence. Conclusion: Ningxue Yishen Granules may induce apoptosis of mesangial cells through regulating bax and bcl-2 gene expression, and promote the dissipation of glomerular proliferative lesions.