孕酮对人牙周膜细胞增殖及成骨分化的调控作用

Progesterone modulates the proliferation and differentiation of the human periodontal ligament cells

目的:通过细胞免疫组织化学方法检测人牙周膜细胞(hPDLCs)中孕激素受体(PR)的表达,研究孕酮在hPDLCs增殖及成骨分化过程中的作用。方法:原代培养hPDLCs,取第4～6代细胞用于实验。分为空白对照组,孕酮组,抑制剂组。用含有不同浓度(1,10,100 nM)孕酮的培养液培养hPDLCs 1,2,3,4,5,6,7 d,采用MTT法检测在孕酮作用下hPDLCs生长曲线的变化情况。检测普通培养液和成骨诱导培养液中的各组细胞在孕酮作用后茜素红染色钙化结节面积的变化。结果:PR在hPDLCs的胞质和胞核中均有表达,且在胞核的表达明显高于胞质。孕酮干预hPDLCs后,细胞的增殖高于对照组,孕激素受体抑制剂能明显抑制hPDLCs的生长。孕酮能够促进茜素红染色钙化结节的形成。在成骨诱导液中,孕酮同样有促进成骨的作用。与普通培养组相比较,孕酮对成骨诱导组的成骨促进作用更为明显。结论:人牙周膜细胞中有PR的表达,孕酮能够促进人牙周膜细胞的增殖和成骨分化。

Objective：To detect the expression of progesterone receptor（PR） in human periodontal ligament cells（hPDLCs） and investigate the proliferation and bone-sparing effects of progesterone.Method：The hPDLCs were stimulated with progesterone（1,10,100 nM） for 1,2,3,4,5,6 and 7 days.MTT assay was performed to assess the effects of progesterone on cell growth curve.To quantify the effects of progesterone on osteogenic differentiation of hPDLCs,alizarin red was performed in basic culture medium and osteogenic medium.Result：The positive signal was strongly expressed in the nuclei but weaker in cytoplasm of hPDLCs.Progesterone can stimulate the proliferation of the hPDLCs,and the PgR antagonist（RU486） can block the effect of progesterone.In both basic culture medium and osteogenic medium,progesterone activated alizarin red-positive nodules.The amount of mineralized matrix increased in the progesterone-treated group in osteogenic medium.Conclusion：Progesterone can stimulate the proliferation and differentiation of the hPDLCs.