二仙汤对骨髓间充质干细胞骨向分化和脂向分化的影响  

Effects of Erxian Decoction on lipid-differentiation and bone-differentiation from bone mesenchymal stem cells

作　　者：刘波[1] 周年 夏循礼[3] 吴文婷 廖夫生[3] 徐彭[3] 张丽[3] 黄思敏 LIU Bo;ZHOU Nian;XIA Xun-li;WU Wen-ting;LIAO Fu-sheng;XU Peng;ZHANG Li;HUANG Si-min(Jiangxi University of Traditional Chinese Medicine,Nanchang 330004,China;Jiangxi Institute for Drug Control,Nanchang 330029,China)

机构地区：[1]江西中医药大学药学院药理教研室,南昌330004 [2]江西省药品检验检测研究院,南昌330029 [3]江西中医药大学,南昌330004

出　　处：《中华中医药杂志》2020年第6期2829-2833,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81560723,No.81860802)。

摘　　要：目的:探讨二仙汤对骨髓间充质干细胞(BMSC)骨向分化和脂向分化的调节作用。方法:制备二仙汤含药血清,作用于P3大鼠BMSC,以MTT法检测大鼠BMSC增殖的作用;分别加入骨向诱导剂和脂向诱导剂,以改良钙钴法和ρNPP法检测碱性磷酸酶活力,以茜素红染色法检测矿化钙结节形成,以油红O染液半定量方法检测脂质累积;以Western Blot法检测骨形成蛋白-2(BMP-2)、骨保护蛋白(OPG)与CCAAT增强子结合蛋白家族(C/EBPα)、过氧化物酶体增殖物激活受体γ(PPARγ)等蛋白表达。结果:二仙汤能促进大鼠BMSC增殖,提高碱性磷酸酶活性及矿化钙结节形成,提高BMP-2和OPG的表达(P<0.05,P<0.01);二仙汤能减少脂质累积(P<0.05,P<0.01),抑制PPARγ和C/EBPα的表达(P<0.01)。结论:二仙汤促进大鼠BMSC骨向分化,抑制大鼠BMSC脂向分化,通过双向调节促进骨形成。Objective:To investigate the regulation effects of Erxian Decoction on lipid and bone-differentiation from bone mesenchymal stem cells(BMSC).Methods:Based on the P3 BMSC isolated from rats,the serum containing Erxian Decoction was prepared for treating.The proliferation of BMSC was detected by MTT.When the osteogenic or adipogenic medium were added respectively,the alkaline phosphatase activity was examined by the improved calcium cobalt method andρNPP method,the formation of mineralized calcium nodules was examined by alizarin red staining,and the accumulation of lipid was also detected by the oil red O dying.The protein expressions of BMP-2,OPG,PPARγand C/EBPαwere detected by Western Blot.Results:The proliferation of BMSC in rats was promoted by the serum containing Erxian Decoction.The activity of alkaline phosphatase of BMSC and the formation of mineralized calcium nodules were increased.The expressions of BMP-2 and OPG were also improved(P<0.05,P<0.01).The accumulation of lipid in BMSC was reduced by the serum containing Erxian Decoction(P<0.05,P<0.01).The expressions of PPARγand C/EBPαwas also decreased(P<0.01).Conclusion:Erxian Decoction promotes the bone-differentiation of BMSCs in rats,inhibits the lipid-differentiation of BMSCs in rats,and promotes bone formation through two-way regulation.

关 键 词：二仙汤 骨向分化 脂向分化 骨髓间充质干细胞 血清药理学 

分 类 号：R285.5[医药卫生—中药学]