尿毒清对晚期糖基化终末产物作用下肾小球系膜细胞影响

目的观察尿毒清对晚期糖基化终末产物（advanced glycosylation end products, AGEs）作用下肾小球系膜细胞的影响,探讨中药尿毒清在治疗糖尿病肾脏疾病（diabetic kidney dis-ease,DKD）中对肾小球系膜细胞的保护作用。方法采用冻干牛血清白蛋白和糖制备无内毒素的AGE-BSA及人工培养牛肾小球系膜细胞,培养时分为3组,在加入终浓度为0.25 mg/ml AGEs 的同时给予浓度为0.2 mg/ml的尿毒清成分溶液为尿毒清组,加入终浓度为0.25 mg/ml AGEs 为 AGEs组,同时设空白对照组,应用 MTT比色法,观察尿毒清组对体外 AGEs 培养的牛肾小球系膜细胞增殖的影响；以系膜细胞与 Alcain Blue（AB,阿利新蓝）结合量为指标,观察尿毒清组对 AGEs导致的系膜细胞膜表面电荷的影响；利用 AO/EB染料观察尿毒清组对 AGEs 诱导作用下的牛肾小球系膜细胞凋亡的影响,并利用荧光显微镜观察尿毒清的抑制作用。结果尿毒清组可以有效地抑制 AGEs对牛肾系膜细胞的促进增殖作用；对抗 AGEs 诱导的牛肾小球系膜细胞表面电荷的影响；减少 AGEs诱导的牛肾小球系膜细胞凋亡。结论尿毒清组能减少对晚期糖基化终末产物作用下肾小球系膜细胞增殖和凋亡,保护肾小球系膜细胞表面电荷。

Influence of Niaoduqing on glomerular mesangial cells with the action of Advanced glycation end products

Objective To observe the effect of Niaoduqing on glomerular mesangial cells with the action of advanced glycosylation end products （AGEs）,and study the protective effect of tradition-al Chinese medicine Niaoduqing on glomerular mesangial cells of the diabetic nephropathy. Methods Endotoxinfree AGE-BSA were prepared by lyophilized bovine serum albumin and glucose, and bovine glomerular mesangial cells were cultured. During cell culture,three groups were set up：Niaoduqing group,given 0.25 mg/ml AGEs and 0.2 mg/mL Niaoduqing component solution simulta-neously;AGEs group,given 0.25 mg/mL AGEs;and blank control group.MTT colorimetric assay was used to observe the effects of Niaoduqing on proliferation of bovine glomerular mesangial cells in vitro cultured with AGEs.By using the binding quantity of mesangial cells and alcain blue （AB）as in-dex,the effects of Niaoduqing on AGEs-induced mesangial cell membrane surface charge were ob-served.The effects of Niaoduqing on AGEs-induced apoptosis of bovine glomerular mesangial cells were examined by AO/EBdye under the fluorescence microscopy.Results Niaoduqingcouldeffective-ly inhibit AGEs-induced proliferation of mesangial cells and reduce AGEs-induced bovine mesangial cell surface charge, and decrease AGEs-induced apoptosis of bovine glomerular mesangial cells.Conclusions Niaoduqing can reduce glomerular mesangial cell proliferation and apoptosis under the action of AGEs,protecting the surface charge on mesangial cells.