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 receptor α (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 ERα antagonist 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, ERα and 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, ERα and 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 ERα and black cohosh medicated serum enhanced the osteoblastic bone formation only via activiation of ERα.

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）