L-精氨酸对人肾系膜细胞增生及胞外基质产生的影响

摘要

AIM: To investigate the effect of L-arginine (L-arg) on the proliferation of human mesangial cells and production of collagen. METHODS: The influence of L-arg on the cell proliferation was determined by MTT assay, immunocytochemical detection of expression of proliferative cell nuclear antigen (PCNA), and flow cytometrical analysis of cell cycle. Procollagen Ⅲ and total collagen level in the supematant and expression of collagen Ⅳ mRNA in human mesangial cells were determined by radioimmunoassay, hydroxyproline colorimetric assay,and reverse transcription polymerase chain reaction (RT-PCR).RESULTS: L-Arg induced inhibition of human mesangial cell lines (HMCL) in a concentrationand time-dependent manner. Immunocytochemical study for PCNA showed the number of cells was decreased,though the percentage of PCNA positive cells was increased in L-arg-treated group. Flow cytometrical analysis showed that cells in S and G2-M phases were markedly increased in L-arg-treated group compared with those in control group. Furthermore, L-arg significantly inhibited the production of procollagen Ⅲ and total collagen in the supernatants determined by radioimmunoassay and hydroxyproline colorimetric assay (P＜0.05 and 0.01, respectively) and inhibited the expression of collagen Ⅳ mRNA determined by RT-PCR (P＜0.01). CONCLUSION: L-arg could exert an inhibitory effect on the proliferation of human mesangial cells and production of rnrnextracellular components, which strongly suggested its potential therapeutic role in the chronic renal scarring.%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G2-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.