续苓健骨汤含药血清对MC3T3-E1细胞分化及增殖的影响

目的探讨续苓健骨汤含药血清对MC3T3-E1成骨细胞分化及增殖的影响。方法制备续苓健骨汤含药血清,实验分为空白对照组、含药血清低剂量组、中剂量组和高剂量组。采用CCK-8法和流式细胞术检测续苓健骨汤含药血清对MC3T3-E1细胞增殖和细胞周期的影响;碱性磷酸酶(ALP)活性测定MC3T3-E1细胞的成骨分化能力;茜素红染色检测MC3T3-E1细胞的矿化能力;实时荧光定量PCR检测成骨分化基因Runx2、OC、Bmp2、Col1a1mRNA水平。结果与空白对照组比较,中、高剂量续苓健骨汤含药血清能促进MC3T3-E1细胞增殖、S期细胞比率和细胞增殖指数,并且呈现一定的剂量依赖性;同时中高剂量续苓健骨汤含药血清组能明显提高MC3T3-E1细胞ALP活性(P0.01)和钙化能力(P0.01),促进Runx2、OC、Bmp2、Col1a1 mRNA的表达(P0.05)。结论续苓健骨汤含药血清能促进成骨细胞MC3T3-E1的增殖,并通过上调骨形成相关基因Runx2、OC、BMP2、Col1a1的表达水平,提高MC3T3-E1细胞的成骨能力。

The effect of Xuling strong bone decoction containing serum on the proliferation and differentiation of MC3T3-E1 cells

Objective To study the effect of Xuling strong bone decoction containing serum on the proliferation and differentiation of MC3T3-E1 cells. Methods The serum containing Xuling strong bone decoction medium was prepared. The experiment was divided into blank control group, drug-containing serum low-dose group, medium-dose group, and high-dose group. CCK-8 method and flow cytometry were used to detect the effect on the proliferation and cell cycle of MC3T3-E1 cells. Alkaline phosphatase (ALP) activity was examined to determine the osteogenic differentiation ability of MC3T3-E1 cells. The mineralization of MC3T3-E1 cells was detected with Alizarin red staining. The expression levels of osteogenic differentiation genes Runx2, OC, BMP2, and Col1a1 were detected using real-time fluorescent quantitative PCR. Results Compared with those in the blank control group, the proliferation, S phase cell ratio, and cell proliferation index of MC3T3-E1 cells were promoted in the low-dose and high-dose group in a dose-dependent manner. ALP activity (P<0.01) and calcification ability (P<0.01) were significantly increased in high-dose group. Also, the mRNA expressions of Runx2, OC, BMP2, and Col1a1 were promoted (P<0.05). Conclusion In a certain concentration range, Xuling strong bone decoction containing serum promotes the proliferation and differentiation of MC3T3-E1 osteoblasts.