健腰密骨片对成骨细胞增殖分化和BMP信号通路报导基因活性的影响

Effect of “Jianyao Migu Tablets” on Osteoblast Proliferation and Differentiation and BMP Signaling Reporter Gene Activity

目的:探讨健腰密骨片对成骨细胞的影响及作用机制。方法:常规培养转染12×SBE-OC-Luc报导基因的成骨细胞株OCT-1细胞(以下简称12×SBE-OC-Luc-OCT1细胞),采用健腰密骨片含药血清进行干预48 h,并以骨形态发生蛋白(BMP)2纯化蛋白(50 ng/ml)为阳性对照。用MTT法检测成骨细胞增殖率,实时荧光定量RT-PCR法检测成骨细胞中Ⅰ型胶原(typeⅠcollagen)、骨钙素(osteocalcin)和BMP2mRNA的表达以及荧光素酶活性检测法检测成骨细胞12×SBE-OC-Luc的活性。结果:与正常对照组比较,健腰密骨片能明显促进成骨细胞的增殖(P<0.05);显著增加成骨细胞中typeⅠcollagen、osteocalcin和BMP2的mRNA表达量(P<0.05);能明显促进成骨细胞中BMP报导基因12×SBE-OC-Luc的荧光素酶活性(P<0.05)。结论:健腰密骨片可通过促进BMP2、typeⅠcollagen和osteocalcin的表达及BMP信号通路报导基因12×SBE-OC-Luc的活性,从而促进成骨细胞的增殖与分化。

Objective： To explore the effects of＂Jianyao Migu Tablets＂and its mechanism on osteoblasts proliferation and differentiation. Methods： The previously established cell lines OCT-1 which transfected with 12 × SBE-OC-Luc reporter gene（ 12 × SBE-OC-Luc-OCT1 cells） were cultured by the regular methods. The serum of rats was intervened by＂Jianyao Migu Tablets＂for 48 h,and with BMP2 purified protein（ 50 ng /ml） as the positive control. The osteoblast proliferation rate was determined by MTT analysis and the mRNA expressions of Type I Collagen,osteocalcin and BMP2 in osteoblasts were detected by RT-PCR. The fluorescence assay was also performed to detect the activity of 12 × SBE-OC-Luc reporter. Results： Compared with the normal control group,＂Jianyao Migu Tablets＂promoted the osteoblast proliferation significantly（ P 〈0. 05）; and the mRNA expressions of type Ⅰ collagen,osteocalcin and BMP2 was enhanced（ P 〈0. 05）; ＂Jianyao Migu Tablets＂promoted the BMP signaling reporter gene 12 × SBE-OC-Luc luciferase activity（ P 〈0. 05）. Conclusion： ＂Jianyao Migu Tablets＂can promote the proliferation and differentiation of osteoblasts by enhancing the expressions of BMP2,Type I Collagen and osteocalcin and by promoting the BMP signaling reporter gene 12 × SBE-OC-Luc luciferase activity.