血小板衍生性生长因子对体外培养人肾小球系膜细胞生长的影响

目的观察血小板衍生性生长因子（ＰＤＧＦ）对体外培养的人肾小球系膜细胞（ＭｓＣ）生长、基质合成和分泌以及ｃ┐ｍｙｃ癌基因表达的影响。方法体外培养的ＭｓＣ培养液中掺入５┐溴脱氧尿嘧啶（ＢｒｄＵ）,采用ＢｒｄＵ单克隆抗体免疫组化法检测ＭｓＣ的增殖情况;应用［３Ｈ］脯氨酸掺入酶消化法测定ＭｓＣ细胞内、外胶原蛋白总量;采用Ｎｏｒｔｈｅｒｎ印迹法检测纤维连接蛋白（ＦＮ）、Ⅳ型胶原、核癌基因ｃ┐ｍｙｃｍＲＮＡ表达结果。结果（１）ＢｒｄＵ掺入法的阳性细胞标记指数在对照组为１９．５％,ＰＤＧＦ组为３４．５％（Ｐ＜０．０１）;（２）ＭｓＣ经ＰＤＧＦ作用后,细胞内、外胶原蛋白量分别为２．６９±０．６０％和３．８７±０．６５％,较正常对照组１．２５±０．５０％和１．６１±０．５１％明显增加（Ｐ＜０．０１）;（３））Ｎｏｒｔｈｅｒｎ印迹法显示ＰＤＧＦ组细胞的ＦＮ、Ⅳ型胶原及ｃ┐ｍｙｃｍＲＮＡ的表达明显高于正常组。结论ＰＤＧＦ不仅可刺激肾小球ＭｓＣ的增殖和胶原蛋白合成,而且可从基因转录水平增加ＦＮ、Ⅳ型胶原及ｃ┐ｍｙｃ癌基因的表达。由此推断ＰＤＧＦ在肾小球疾病的发生、发展中可能起着重要作用

Effect of platelet derived growth factor on the growth of human glomerular mesangial cells in vitro

OBJECTIVE: To study the effect of plateletderived growth factor (PDGF) on the growth of human glomerular mesangial cells (MsC) in vitro. METHODS: Immunohistochemistry for the detection of BrdU positive cells; 3H] proline incorporation for the bioactive assay of total amount of collagen; and Northern blot analysis for the determination of fibronectin (FN) alpha 1 (IV) collagen and c-myc mRNA expression. RESULTS: The cell labelling index (LI) of BrdU incorporation in the PDGF group is 34.5% higher than that in the normal group, which was 19.5% (P < 0.01), The total amount of collagen secreted by MsC within cells and outside of cells in the PDGF group were 2.69 +/- 0.60% and 3.87 +/- 0.65% respectively, much higher than that of the normal group, which were 1.25 +/- 0.50% and 1.61 +/- 0.51% respectively (P < 0.01). Northern blot analysis revealed an increase of FN, Col IV, c-myc mRNA levels in the PDGF group. CONCLUSION: PDGF can stimulate cell proliferation, synthesis of collagen and enhance FN, Col IV, c-myc mRNA expression. It is possible that PDGF may play an important role in the development and progression of glomerulopathy.