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目的探讨ｒａｆ１基因高表达对肾小球系膜细胞增殖的调控作用。方法应用细胞直接计数、流式细胞仪及特异底物磷酸化法分别检测ｒａｆ１基因转染细胞的增殖率、细胞周期分布及与细胞外信号调节的蛋白酶（ＥＲＫ）和ｃｊｕｎ蛋白酶（ＪＮＫ）的活性。结果ｒａｆ１基因高表达细胞早期生长正常，但于后期生长迟缓，细胞数目（２２．０±２．５）×１０５个／孔与对照组（３３．４±２．７）×１０５个／孔相比较显著减少（Ｐ＜０．０５）；细胞于生长第５天时停滞于Ｇ０／Ｇ１期（８１．３±３．７），（６２．３±２．４）％，（Ｐ＜０．０１），而Ｓ期细胞数量（８．３±０．９）％明显少于对照组（２７．０±１．５）％，（Ｐ＜０．０１）；其ＥＲＫ活性［（２１４±１６）ｐｍｏｌ·ｍｇ－１·ｍｉｎ－１］显著高于对照组［（８２±９）ｐｍｏｌ·ｍｇ－１·ｍｉｎ－１，Ｐ＜０．０５］，但ＪＮＫ活性则较对照组明显降低约５０％。结论在鼠肾小球系膜细胞中，Ｒａｆ１蛋白激酶确可作为ＥＲＫ的上游信号激活细胞增殖信号，但其持续高表达则能导致细胞生长周期停滞，这一作用很可能与ＪＮＫ活性抑制或细胞周期调控蛋白的变化有关。
Objective To investigate the regulatory effect of raf1 gene overexpression on mesangial cell proliferation. Methods Using cell direct counting, flow cytometry and specific substrate phosphorylation were detected raf 1 gene transfected cells proliferation rate, cell cycle distribution and extracellular signal-regulated proteinase (ERK) and c jun protease ( JNK) activity. Results The cells with high expression of raf1 gene grew normally at early stage, but grew slowly at the later stage. The number of cells (22.0 ± 2.5) × 105 cells / well and control group (33.4 ± 2.7) × 105 cells / (P <0.05). The cells were arrested in G0 / G1 phase (81.3 ± 3.7), (62.3 ± 2.4)% on day 5 .01), while the number of S phase cells (8.3 ± 0.9)% was significantly lower than that of the control group (27.0 ± 1.5)%, (P <0.01) 16) pmol · mg-1 · min-1] was significantly higher than that of the control group [(82 ± 9) pmol · mg-1 · min-1, P <0.05] 50%. Conclusions Raf1 protein kinase can activate the cell proliferation signal as the upstream signal of ERK in rat glomerular mesangial cells, but sustained high expression of Raf1 protein kinase can lead to cell cycle arrest, which may be related to the inhibition of JNK activity Or changes in cell cycle regulatory proteins.