论文部分内容阅读
目的:研究L-精氨酸(L-arg)对人肾系膜细胞增生和胞外基质成分胶原产生的影响。方法:采用MTT法、细胞免疫化学检测增殖细胞核抗原(PCNA)表达以及流式细胞仪等方法观察L-arg对系膜细胞增殖的影响;L-arg对系膜细胞胞外基质产生的影响分别采用放免法、羟化脯氨酸比色法以及RT-PCR等方法测定前胶原Ⅲ、总胶原以及胶原Ⅳ mRNA的表达。结果:L-arg呈剂量和时间依赖性抑制人系膜细胞增殖;细胞免疫化学法显示L-arg导致细胞总数下降,但PCNA阳性比例相对增高,用流式细胞仪进一步证实L-arg治疗组细胞主要处于细胞周期的S及G_2-M期。另外,L-arg显著抑制培养的系膜细胞上清液中总胶原和前胶原Ⅲ合成(分别为P<0.05和P<0.01)以及细胞对胶原Ⅳ mRNA的表达(P<0.01)。结论:L-arg可抑制人肾系膜细胞增生和胞外基质成分产生,此提示L-arg对慢性肾脏纤维化可能有潜在的治疗价值。
Objective: To investigate the effect of L-arginine on the proliferation of human mesangial cells and the collagen production of extracellular matrix. Methods: The effect of L-arg on the proliferation of mesangial cells was observed by MTT assay, the expression of proliferating cell nuclear antigen (PCNA) by flow cytometry and flow cytometry. The effects of L-arg on the production of extracellular matrix The expressions of procollagen Ⅲ, total collagen and collagen Ⅳ mRNA were detected by radioimmunoassay, hydroxyproline colorimetry and RT-PCR. Results: L-arg inhibited the proliferation of human mesangial cells in a dose- and time-dependent manner. Immunocytochemistry showed that L-arg resulted in a decrease in the total number of cells but a positive proportion of PCNA. Flow cytometry further confirmed that the L-arg treatment group Cells are mainly in the cell cycle S and G_2-M phase. In addition, L-arg significantly inhibited the total collagen and procollagen III synthesis (P <0.05 and P <0.01, respectively) and the collagen IV mRNA expression (P <0.01) in cultured mesangial cell supernatant. CONCLUSION: L-arg inhibits the proliferation of human mesangial cells and the production of extracellular matrix components, suggesting that L-arg may have potential therapeutic value in chronic kidney fibrosis.