中国骨质疏松杂志

芍药苷促进小鼠成骨分化抗骨质疏松作用的实验研究

Experimental study on the effect of paeoniflorin in promoting osteogenic differentiation and anti-osteoporosis in mice

DOI：10.3969/j.issn.1006-7108.2020.12.005

中文关键词: 芍药苷 MC3T3-E1细胞成骨分化卵巢摘除术

英文关键词:paeoniflorin MC3T3-E1 cells osteogenic differentiation ovariectomy

基金项目:辽宁省自然科学基金指导计划（2019-ZD-0761）；辽宁省教育厅科学研究课题（JC2019016）

作者	单位
杨立宇郭然牟帅张一奇杨礼庆付勤*	中国医科大学附属盛京医院骨科，辽宁沈阳 110003

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中文摘要:

目的探讨芍药苷对MC3T3-E1成骨细胞分化以及小鼠骨质疏松模型的影响。方法体外细胞实验分为对照组、不同剂量芍药苷干预组。通过CCK-8法检测芍药苷对MC3T3-E1成骨细胞活力的影响；采用碱性磷酸酶（ALP）染色以及活性检测芍药苷促进MC3T3-E1成骨分化能力；通过茜素红染色检测芍药苷促矿化能力；运用荧光定量PCR、Western blot检测Runx2、OPG、RANKL、Col1α1的mRNA以及蛋白表达情况。选取8周龄C57BL/6小鼠24只，分为假手术组、骨质疏松模型组、药物干预组。选取各小鼠左侧股骨远端以及胫骨近端的区域进行苏木素-伊红（HE）染色、Runx2免疫组织化学以及micro-CT扫描。结果中、高剂量芍药苷干预组可以促进MC3T3-E1细胞ALP的活性（P<0.05）；高剂量芍药苷能够促进MC3T3-E1细胞的矿化能力（P<0.01）；同时中、高剂量能够促进OPG、Runx2蛋白的表达（P<0.05），抑制RANKL的表达（P<0.05）。与假手术组比较，OVX组骨微结构破坏明显，Runx2蛋白表达显著减少（P<0.01）；芍药苷干预后骨质疏松模型小鼠骨小梁数量以及厚度显著增加（P<0.01），骨小梁间隔减少明显（P<0.01），Runx2蛋白提升明显（P<0.01）。结论芍药苷能够促进成骨细胞的分化，上调成骨分化基因，改善骨质疏松模型小鼠的骨微结构，具有抗骨质疏松治疗的潜在价值。

英文摘要:

Objective To investigate the effect of paeoniflorin on osteoblastic differentiation of MC3T3-E1 and osteoporosis in mice. Methods In vitro experiments cells were divided into control group and different concentrations of paeoniflorin intervention groups. CCK-8 method was used to detect the effect of paeoniflorin on the activity of MC3T3-E1 osteoblasts. Alkaline phosphatase (ALP) staining and activity detection of paeoniflorin effects on the osteogenic differentiation of MC3T3-E1 were utilized. Alizarin red staining was used to detect the effect of paeoniflorin on the mineralization capacity of MC3T3-E1. MRNA and related proteins such as Runx2, OPG, RANKL, Col1α1 were detected by quantitative fluorescence PCR and Western blot. Twenty-four 8-week-old C57BL/6 mice were randomly divided into three groups, namely Sham group, OVX model group and OVX+Pa model intervention group. After 2 months of intervention, the left distal femur and proximal tibia regions of each mouse were selected for hematoxylin-eosin (HE) staining, Runx2 immunohistochemistry, and micro-CT scanning. Results Compared with the control group, medium and high dose paeoniflorin group could promote the activity and expression of ALP in MC3T3-E1 cells (P<0.05). High dose paeoniflorin could promote the mineralization ability of MC3T3-E1 cells (P<0.01). Meanwhile, paeoniflorin could promote the expression of osteogenic differentiation genes OPG and Runx2(P<0.05), and inhibited the expression of RANKL gene and protein (P<0.05). In vivo, results showed that OVX group induced significant bone microstructure destruction and significantly decreased Runx2 protein expression compared with Sham group (P<0.01). The number and thickness of bone trabeculae were significantly attenuated in the paeoniflorin group (P<0.01),the bone trabeculae space was significantly decreased (P<0.01), and the expression of Runx2 protein was increased compared with the osteoporotic mice (P<0.01). Conclusion Paeoniflorin can promote the differentiation of osteoblasts, elevate the expression of related bone formation genes, improve the bone microstructure of osteoporosis model mice. It has the potential value of anti-osteoporosis treatment.

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