肝素和PDGF对人肾小球系膜细胞基质合成和分泌的影响

Effect of heparin and PDGF on synthesis and secretion of extracellular matrix in human cultured mesangial cell

目的：探讨肝素和血小板源性生长因子（ＰＤＧＦ）对体外培养的人肾小球系膜细胞（ＭｓＣ）胶原合成和分泌以及纤连蛋白（ＦＮ）、α１（ＩＶ）型胶原ｍＲＮＡ表达的影响。方法：应用３Ｈ－脯氨酸掺入、胶原酶消化法测定胶原含量；采用Ｎｏｒｔｈｅｒｎｂｌｏｔ法检测ＦＮ、α１（ＩＶ）型胶原ｍＲＮＡ。结果：对照组、肝素组及ＰＤＧＦ组细胞内胶原含量分别为（１．２５０±０．５０１）％，（０．８０９±０．１６１）％和（２．６９０±０．５９７）％，而其培养液中的含量分别为（１．６０８±０．５１２）％，（０．１２２±０．０８９）％和（３．８７０±０．６４７）％。上述结果显示肝素对该细胞合成和分泌胶原有抑制作用（Ｐ＜０．０５，Ｐ＜０．０１），而ＰＤＧＦ则具有明显促进作用（Ｐ均＜０．０１）。Ｎｏｒｔｈｅｒｎｂｌｏｔ分析结果显示肝素可抑制ＭｓＣＦＮ、α１（ＩＶ）型胶原ｍＲＮＡ的表达，而ＰＤＧＦ则可明显增强其表达。结论：肝素对人肾小球ＭｓＣ基质的合成和分泌有抑制作用，而ＰＤＧＦ则具有促进作用，其作用机制均发生在基因转录水平上。

Purpose To study the effect of heparin and plateletderived growth factor(PDGF) on synthesis and secretion of total collagen and expression of fibronectin (FN), α1 (IV) collagen mRNA in human cultured mesangial cells (MsC). Methods 3Hproline incorporation with collagenase digestion for mesurement of total collagen, and Northern blot analysis for detection of FN, α1 (Ⅳ) collagen mRNA. Results The total amount of collagen, newly formed and secreted by MsC were (1250±0501)%, (0809±0161)%, (2690±0597)% and (1608±0512)%, (0122±0089)%, (3870±0647)% in control, heparin and PDGF groups, respectively. Collagen values in heparin group were significantly decreased compared with normal group (P<005 or P<001), but increased in PDGF group (P<001). Northern blot analysis revealed that the expression of FN, α1(Ⅳ) collagen mRNA on MsC in heparin group were inhibited as compared to normal serum group, but enhanced in PDGF group. Conclusions The ECM synthesis and secretion are inhibited by heparin in human MsC, but stimulated by PDGF.