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目的:研究当归补血汤对培养在高糖条件下肾小球系膜细胞(GMC)增殖及GMC中转化生长因子-β1mRNA(TGF-β1mRNA)及核转录因子(NF-кB)蛋白的表达的影响,探讨其在糖尿病肾病(DN)防治中的意义。方法:①利用噻唑蓝(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;②用荧光定量PCR的方法检测各种处理因素的作用下各组GMC中TGF-β1mRNA表达的情况;③用Western-blot的方法检测各种处理因素的作用下各组GMC中NF-кB蛋白的表达情况。结果:高糖能明显刺激体外培养的GMC增殖,同时细胞中TGF-β1mRNA及NF-кB蛋白表达也增加(P<0.05)。而当归补血汤能明显抑制GMC增殖及细胞中TGF-β1mRNA及NF-кB蛋白表达也相应减少(P<0.05或P<0.01),并呈一定量效关系,即随糖或药物浓度的增加,GMC增殖及细胞中TGF-β1mRNA及NF-кB蛋白表达也相应增加或减少。结论:高糖可促进GMC增殖及GMC中TGF-β1mRNA和NF-кB蛋白的表达增加,当归补血汤能明显抑制高糖条件下GMC增殖,抑制细胞中TGF-β1mRNA及NF-кB蛋白表达增加,说明当归补血汤能直接抑制GMC增殖、GMC中TGF-β1mRNA及NF-кB蛋白表达增加,从而发挥预防和治疗肾小球硬化和间质纤维化的发生、发展,进而延缓DN的进展。
Objective: To study the effect of Dangguibuxuetang on the proliferation of glomerular mesangial cells (GMCs) and the expression of TGF-β1mRNA and NF-κB in GMCs under high glucose conditions , To explore its significance in the prevention and treatment of diabetic nephropathy (DN). Methods: ① MTT assay was used to observe the proliferation of GMCs under various treatment conditions; ② The expression of TGF-β1 mRNA in GMCs was detected by fluorescence quantitative PCR ; ③ Western-blot method was used to detect the expression of NF-кB in GMCs under various treatment factors. RESULTS: High glucose significantly stimulated the proliferation of GMC in vitro and the expression of TGF-β1mRNA and NF-κB protein also increased (P <0.05). However, Dangguibuxuetang could significantly inhibit the proliferation of GMC and decrease the expression of TGF-β1mRNA and NF-κB protein (P <0.05 or P <0.01) in a dose-dependent manner. With the increase of glucose or drug concentration, GMC proliferation and cell TGF-β1 mRNA and NF-κB protein expression corresponding increase or decrease. Conclusion: High glucose can promote the proliferation of GMC and the expression of TGF-β1mRNA and NF-κB protein in GMC. Dangguibuxuetang can obviously inhibit the proliferation of GMC under high glucose and inhibit the expression of TGF-β1mRNA and NF- It indicates that Dangguibuxuetang can directly inhibit the proliferation of GMC, and increase the expression of TGF-β1mRNA and NF-κB in GMC, so as to prevent and treat the occurrence and development of glomerulosclerosis and interstitial fibrosis and further delay the progression of DN.