当归补血汤对高糖条件下系膜细胞增殖及细胞外基质表达的影响

Effects of Astragalus and Angelica Mixture on the Proliferation of Glomerular Mesangial Cells and Expression of Extracellular Matrix Cultured in High Glucose

目的:研究当归补血汤对培养在高糖条件下系膜细胞(GMC)增殖及细胞上清液中层粘连蛋白(LN)、Ⅳ胶原纤维(ColⅣ)、纤维连接蛋白(FN)的表达的影响,探讨其在糖尿病肾病(DN)防治中的意义。方法:①采用酶联免疫吸附试验(ELISA)测定经体外培养的大鼠肾小球系膜细胞在各种处理因素的作用下上清液中LN、Ⅳ型胶原及FN的表达情况;②利用四甲基偶氮唑蓝(MTT)比色法动态观察各种处理因素的作用下各组GMC增殖情况。结果:高糖能明显刺激体外培养的GMC增殖,同时促进GMC分泌FN、LN和ColⅣ(P<0.05或P<0.01)。而当归补血汤能抑制GMC增殖及GMC分泌FN、LN和ColⅣ,并呈一定量效关系(P<0.05或P<0.01),即随糖或药物浓度的增加,GMC增殖、GMC分泌FN、LN和ColⅣ的能力也相应增加或减少。结论:高糖可促进GMC增殖、细胞外基质(ECM)蛋白分泌,当归补血汤能明显抑制高糖条件下GMC增殖、及其分泌FN、LN和ColⅣ,说明当归补血汤能直接抑制GMC增殖及分泌ECM,从而发挥预防肾小球纤维化及硬化的发生、发展,进而延缓DN的进展。

Objective： To investigate the effect of astragalus and angelica mixture（Angelicae Sinensis Decoction for Supplementing Blood,a traditional herbal remedies of China） on glomerular mesangial cell（GMC） proliferation and the expression of laminin（LN）,collagen type Ⅳ（ColⅣ）,fibronectin（FN） in vitro under high glucose.Methods： ①Enzyme linked immunoadsordent assay（ELISA） was adopted to determine the expression of LN,ColⅣ and FN in cultured rat glomerular mesangial cells;②The thiazolyl blue tetrazolium bromide（MTT） colorimetric method was used to observe the glomerular mesangial cell proliferation. Results： In vitro cultured glomerular mesangial cells were obviously proliferating under the stimulation of high glucose,which also promote LN,ColⅣand FN secretion by glomerular mesangial cells（P〈0.05 or P〈0.01）;but astragalus and angelica mixture can inhibit GMCs proliferating and secreting LN,ColⅣ and FN in a dose-effect manner.Conclusion： High glucose can promote GMC proliferation and secretion of extracellular matrix, but Angelicae Sinensis Decoction for Supplementing Blood could inhibit it,which could be helpful for preventing of the progression of diabetic nephropathy.