朝藿定A对原代骨髓间充质干细胞向成骨细胞分化的作用  

Effect of epimedin A on the differentiation of primary bone marrow mesenchymal stem cells into osteoblasts in vitro

作　　者：刘颖[1] 柴丽娟[2] 黄菊阳 王琴 宋蕾 周昆[2] 李云章[1] LIU Ying;CHAI Lijuan;HUANG Juyang;WANG Qin;SONG Lei;ZHOU Kun;LI Yunzhang(College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China;Key Laboratory of Chinese Medicine Pharmacology,Institute of Traditional Chinese Medicine,Tianjin University of TCM,Tianjin 300193,China)

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]天津中医药大学中医药研究院,天津市中药药理重点实验室,天津300193

出　　处：《中国兽医学报》2021年第5期980-984,991,共6页Chinese Journal of Veterinary Science

基　　金：国家重大新药创制专项课题基金资助项目(2011ZX09201-201-21,2014ZX09304307-001-005)。

摘　　要：为了研究朝藿定A对原代骨髓间充质干细胞(bone marrow mesenchymal stem cells, BMSCs)向成骨细胞分化的机制,本试验采用正常小鼠骨髓来源的间充质干细胞,通过观察朝藿定A对骨髓间充质干细胞成骨分化、成脂分化和矿化能力的影响,应用ELISA技术,研究其抗骨质疏松作用及相关分子机制。结果显示,朝藿定A给药3 d后,成骨诱导后的BMSCs分泌了更多碱性磷酸酶(alkaline phosphatase, ALP),在朝藿定A给药14 d后,矿化结节显著增多,朝藿定A给药10 d后,朝藿定A低剂量组成脂分化的程度没有明显变化,而高剂量组成脂分化较对照组显著降低。BMSCs经成骨诱导及朝藿定A给药7 d后,给予0.01μmol/L朝藿定A时,上清中骨保护素(osteoprotegerin, OPG)水平显著升高,给予0.1 nmol/L朝藿定A时,成骨细胞表达核转录因子(receptor activator for nuclear factor-κB ligand, RANKL)水平呈上升趋势;而给予1,100μmol/L朝藿定A时,RANKL水平呈下降趋势。从OPG/RANKL的结果来看,给予0.1 nmol/L朝藿定A时OPG/RANKL值较对照组显著升高。说明朝藿定A可以促进BMSCs向成骨细胞分化并抑制其成脂分化,可以促进成骨细胞矿化并提高成骨细胞OPG/RANKL水平从而发挥骨保护作用。To study the mechanism of epimedin A on the differentiation of primary bone marrow mesenchymal stem cells into osteoblasts, mesenchymal stem cells from normal mouse bone marrow were used to observe the effects of epimedin A on bone formation differentiation, adipogenic differentiation and mineralization of bone marrow mesenchymal stem cells, and ELISA was used to study its anti-osteoporosis effect and related molecular mechanisms.Three days after the administration of epimedin A,BMSCs secreted more alkaline phosphatase(ALP) after osteogenic induction.Fourteen days after the administration of epimedin A,the mineralized nodules increased significantly.After 10 d of administration of epimedin A,the degree of adipogenic differentiation in the low-dose group did not change significantly, while the lipid differentiation of high-dose group was significantly lower than that of the control group.Seven days after BMSCs were induced by osteogenic and treated with epimedin A,the OPG level in the supernatant was significantly increased when 0.01 μmol/L epimedin A was administered.When 0.1 nmol/L epimedin A was administered,the RANKL level was increased.The RANKL level showed a downward trend when given 1,100μmol/L epimedin A.The results of OPG/RANKL showed that the value of OPG/RANKL was significantly higher than that of the control group when 0.1 nmol/L epimedin A was given.This study shows that epimedin A can promote the differentiation of BMSCs into osteoblasts and inhibit their adipogenic differentiation,promote osteoblast mineralization and increase the OPG/RANKL levels of osteoblasts to exert bone protection.

关 键 词：朝藿定A 骨髓间充质干细胞 成骨细胞 

分 类 号：S859.7[农业科学—临床兽医学]