基于大麻素2型受体和雌激素受体α研究升麻和黑升麻含药血清对成骨细胞功能的影响  

Effects of Cimicifuga foetida and black cohosh medicated serum on osteoblasts functions based on CB2R and ERα

作　　者：赵子慧 胡思婧 陈高策 叶欣园 刘燕 张泉龙 徐金龙 秦路平 韩婷 张巧艳 ZHAO Zi-hui;HU Si-jing;CHEN Gao-ce;YE Xin-yuan;LIU Yan;ZHANG Quan-long;XU Jin-long;QIN Lu-ping;HAN Ting;ZHANG Qiao-yan(School of Pharmaceutical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China;969 Hospital of PLA Joint Logistics Support Forces,Hohhot 010051,China;School of Pharmacy,Naval Medical University,Shanghai 200433,China)

机构地区：[1]浙江中医药大学药学院,浙江杭州310053 [2]中国人民解放军联勤保障部队第九六九医院,内蒙古呼和浩特010051 [3]中国人民解放军海军军医大学药学院,上海200433

出　　处：《中草药》2023年第18期5941-5951,共11页Chinese Traditional and Herbal Drugs

基　　金：上海市科委科技支撑计划项目(22S21901600);国家自然科学基金资助项目(81973534)。

摘　　要：目的研究升麻Cimicifuga foetida和黑升麻含药血清基于大麻素2型受体(cannabinoid receptor type 2,CB2R)与雌激素受体α(estrogen receptorα,ERα)对成骨细胞骨形成的调控作用。方法采用高分辨质谱法鉴定升麻和黑升麻中的化学成分;升麻与黑升麻含药血清或与CB2R反向激动剂AM630或ERα抑制剂ICI联合处理MC3T3-E1成骨细胞,CCK-8法测定细胞增殖活性;试剂盒测定碱性磷酸酶(alkaline phosphatase,ALP)活性;茜素红染色观察成骨细胞骨矿化结节的形成;Western blotting检测骨形成相关蛋白及CB2R与ERα的表达。结果升麻和黑升麻中分别鉴定了12、14个化学成分,其中有3个共有成分。升麻和黑升麻含药血清可显著增加成骨细胞的增殖、ALP活性和骨矿化结节形成,增加CB2R和ERα及骨形成相关蛋白的表达。15%升麻含药血清对MC3T3-E1成骨细胞增殖、ALP活性、骨矿化结节形成及骨形成相关蛋白表达的促进作用可被AM630和ICI所逆转,15%黑升麻含药血清对MC3T3-E1细胞骨形成的促进作用仅可被ICI所逆转。结论升麻含药血清可通过CB2R与ERα双靶点促进成骨细胞的骨形成,黑升麻含药血清仅通过激活ERα增加成骨细胞的骨形成。Objective To study the regulatory effects of Cimicifuga foetida and black cohosh medicated serum on osteoblastic bone formation based on cannabinoid receptor type 2(CB2R)and estrogen receptorα(ERα).Methods High-resolution mass spectrum was used to identify the chemical constituents in C.foetida and black cohosh.The osteoblastic MC3T3-E1 cells were treated with various concentrations C.foetida and black cohosh medicated serum or the combination with CB2R inverse agonist AM630 or ERαantagonist ICI.CCK-8 method was used to determine the proliferation of MC3T3-E1 cells.The assay kit was used to measure the alkaline phosphatase(ALP)activity.The alizarin red staining was used to detect the formation of bone mineralized nodules in MC3T3-E1 cells.Western blotting was applied to analyze the expressions of CB2R,ERαand related proteins with osteoblastic bone formation.Results A total of 12 chemical constituents were identified in C.foetida,and 14 constituents in black cohosh,with three common constituents.Both C.foetida and black cohosh medicated serum significantly increased the proliferation,ALP activity and the formation of bone mineralized nodules,and also enhanced the expression of CB2R,ERαand related proteins associated with osteoblastic bone formation.Moreover,the promoting effects 15%C.foetida medicated serum on osteoblastic MC3T3-E1 cells could be reversed by AM630 and ICI,while the improvement effects of 15%black cohosh medicated serum on osteoblast only could be reversed by ICI.Conclusion C.foetida medicated serum increased the osteoblastic bone formation via activation of CB2R and ERαand black cohosh medicated serum enhanced the osteoblastic bone formation only via activiation of ERα.

关 键 词：升麻 黑升麻 成骨细胞 大麻素2型受体 雌激素受体Α 升麻消旋体A 7 8-二羟基升麻醇 2′-O-acetyl cimicifugoside H-1 

分 类 号：R285.5[医药卫生—中药学]