摘要
目的:体外研究高糖对人肾脏间质成纤维细胞c-met表达的影响,并观察黄芪药物血清对其的调节作用。方法:体外培养人肾脏间质成纤维细胞,按培养液不同分为正常对照组、高糖组和甘露醇组。于作用后6、12、24、48和96h,采用逆转录-聚合酶链反应方法检测c-met和转化生长因子β1(transforming growth factor-beta1,TGF-β1)mRNA表达,蛋白印迹分析检测c-met蛋白的表达。制备含药血清,将成纤维细胞分高糖组、对照血清组和10%黄芪含药血清组,作用24h和48h后,检测c-met蛋白和mRNA表达情况。结果:与正常对照组比较,高糖组细胞培养12h时c-met mRNA表达增加(P<0.05),24h达高峰(P<0.01),之后表达逐渐下降;高糖组细胞培养24h时,TGF-β1的表达开始增加(P<0.05),96h达到高峰(P<0.01)。与高糖组以及对照血清组比较,黄芪含药血清可使细胞c-met mRNA和蛋白表达增加(P<0.01,P<0.05)。结论:高糖刺激早期诱导c-met表达,之后表达下降。黄芪可以通过诱导c-met表达延缓糖尿病肾病的进展。
Objective： To investigate the effect of high glucose on the expression of c-met in human kidney fibroblasts in vitro, and to explore the regulation of Radix Astragali.
Methods： A cell culture system of human kidney fibroblasts was developed in vitro, The human kidney fibroblasts were divided into normal control group, high glucose group and mannitol group. Expressions of c-met and transforming growth factor-betal （TGF-β1） mRNAs were detected by reverse transcription polymerase chain reaction （RT-PCR） and the expressions of c-met protein were analyzed by Western blot method after 6-, 12-, 24-, 48- and 96-hour culture. The human kidney fibroblasts were also cultured with 10 % Radix Astragati containing serum; the expressions of c-met mRNA and protein were detected after 24- and 48-hour culture. Results： Compared with the normal control group, expression of c-met mRNA in the high glucose group was significantly increased after 12-hour culture （P〈0. 05）, arriving at the peak after 24-hour culture （P〈0. 01）. The level of TGF-β1 mRNA was higher in the high glucose group than that in the normal control group after 24- hour culture （ P〈0. 05）, arriving at the peak after 96-hour culture （ P〈0. 01）. Forty-eight hours after treating with 10% Radix Astragali containing serum, the levels of c-met mRNA and protein in fibroblasts were increased, and were higher than those in the high glucose group （P〈0. 01, P〈0. 05）.
Conclusion： High glucose can induce the expressions of c-met mRNA and protein in earlier period, and then inhibit the expressions. Radix Astragali can up-regulate the expressions of c-met mRNA and protein of human kidney fibroblasts, which may be one of its action mechanisms in delaying the progression of diabetic nephropathy.
出处
《中西医结合学报》
CAS
2008年第5期482-487,共6页
Journal of Chinese Integrative Medicine
基金
上海市优秀青年医学人才培养计划[No沪卫人(2004)80]
