补肾法防治原发性骨质疏松症的细胞、分子作用机理研究

Cellullar and molecular effective mechanism of kidney invigoration on primary osteoporosis

目的:在骨细胞及分子水平观察了补肾法防治骨质疏松作用环节。方法:以建立的成骨细胞、破骨细胞及骨髓基质细胞培养体系进行体外实验,采用MTT比色分析法及3H-胸腺嘧啶核苷渗入法测定成骨细胞增殖及分化;采用流式细胞仪观察成骨细胞的凋亡;采用原位分子杂交研究TGF-β1、MMP-9 mRNA的表达。结果:补肾法可促进成骨细胞的增殖及分化;部分保护由肿瘤坏死因子α诱导的成骨细胞凋亡,使成骨细胞转化生长因子β1(TGF-β1)mRNA的表达增强;抑制破骨细胞的骨吸收活性;降低破骨细胞基质金属蛋白酶9(MMP-9)mRNA的表达。结论:补肾法具有抑制骨吸收,促进骨形成的双重药效作用,其机理部分是在mRNA水平上,通过对骨细胞内的TGF-β1、MMP-9等影响骨代谢的细胞因子及蛋白酶的调控而达到的。

Objective：To study the effective mechanism of kidney invigoration on osteoporosis in bone cell and molecular level respectively.Methods：This study took the culture systems of osteoblast,osteoclast,and marrow matrical cell were used in vitro.Chalorimetric MMT,3H-thymidine incorporation was detected to determine the condition of cell prolifera tion and differentiation.The flow cytometry was used to determine the condition of the osteoblastic cell apoptosis.And using in situ hybridization on cultured cell studied the expression of TGF-β1 mRNA in osteoblasts and MMP-9 mRNA inosteoclasts.Results：Kindney invigoration might promote proliferate of osteoblast,protect apoptosis of osteoporostic cell induced by TNF-α,boost up the expression of mRNA in TGF-β1,prohibit bone resorption activity of osteoclast,reduce mRNA expression of MMP-9 osteoclast.Conclusions：It indicated that kidney invigoration not only can depress bone resorption,but also improve bone formation,which means it has dual pharmic effect.The partial mechanism of preventing and treating osteoporosis with kidney invigoration are which might regulate some cyctokines and protein enzymes such as TGF-β1,IL-6,MMP-9 in cell at mRNA level.