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

作　　者：杨立宇[1] 郭然[1] 牟帅[1] 张一奇[1] 杨礼庆[1] 付勤[1] YANG liyu;GUO ran;MU Shuai;ZHANG yiqi;YANG liqing;FU Qin(Department of Orthopaedic, Shengjing Hospital of China Medical University, Shenyang 110003,China)

机构地区：[1]中国医科大学附属盛京医院骨科,辽宁沈阳110003

出　　处：《中国骨质疏松杂志》2020年第12期1742-1748,1759,共8页Chinese Journal of Osteoporosis

基　　金：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)。

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。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,i

关 键 词：芍药苷 MC3T3-E1细胞 成骨分化 卵巢摘除术 

分 类 号：R285.5[医药卫生—中药学]