艾灸通过调控肠道菌群相关血清素代谢抑制绝经后骨丢失  

Moxibustion Inhibits Postmenopausal Bone Loss by Regulating the Metabolism of Gut Microbiota-Related Serotonin

作　　者：黄显皓 Camilo Alberto Pinzon Galvis 胥惠民 鲁凌云 杨晗 李宁[1] 文谦[1] HUANG Xianhao;Camilo Alberto Pinzon Galvis;XU Huimin;LU Lingyun;YANG Han;LI Ning;WEN Qian(Division of Internal Medicine,Institute of Integrated Traditional Chinese and Western Medicine,West China Hospital,Sichuan University,Chengdu 610041,China)

机构地区：[1]四川大学华西医院中西医结合中心中西医结合内科,成都610041

出　　处：《四川大学学报(医学版)》2025年第1期129-136,共8页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(No.82204847);四川省科技厅重点研发项目(No.2023YFS0332)资助。

摘　　要：目的探究中医艾灸疗法对卵巢切除的雌激素缺乏小鼠骨代谢调控作用并探究其机制。方法将12周龄雌性C57BL/6J小鼠随机分为假手术组(SHAM)、卵巢切除术组(OVX)、卵巢切除术合并广谱抗生素干预组(OVX-A)、卵巢切除术后艾灸干预组(OVX-M)及卵巢切除术后广谱抗生素干预同时艾灸组(OVX-A-M),每组5只。手术后4周,分别给予广谱抗生素和(或)艾灸干预4周后处死取材。采用Micro-CT检测各组小鼠股骨远端骨量并进行参数分析〔包括骨体积分数(bone volume/tissue volume,BV/TV)、骨小梁数目(trabecular number,Tb.N)、骨小梁厚度(trabecular thickness,Tb.Th)、骨小梁分离度(trabecular separation,Tb.Sp)、皮质骨厚度(cortical thickness,Ct.Th)〕。采用色氨酸靶向代谢组学检测血清色氨酸代谢物水平,qPCR检测骨髓血清素(serotonin,SER)受体Htr2a和Htr2b mRNA水平。此外,采用不同浓度梯度(0、0.01、0.1、1、10μmol/L)血清素干预原代骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs),采用碱性磷酸酶(ALP)染色观察BMSCs成骨分化情况,同时采用qPCR检测细胞成骨相关基因Colla1及血清素受体Htr2a和Htr2b mRNA水平。结果与SHAM小鼠相比,OVX小鼠骨微结构破坏,BV/TV降低〔(10.57±2.82)%vs.(4.20±0.96)%,P<0.01〕,Tb.N减少〔(3.16±0.11)vs.(2.25±0.15),P<0.01〕,Tb.Sp增加〔(0.31±0.01)vs.(0.45±0.03),P<0.01〕,血清色氨酸代谢物SER水平降低;与OVX小鼠相比,艾灸干预的OVX-M小鼠骨微结构改善,BV/TV增加到(7.51±1.42)%(P<0.05),血清SER水平及骨髓Htr2a基因表达水平升高(P<0.05)。而给予广谱抗生素干扰肠道菌群后,艾灸逆转OVX小鼠骨丢失的作用消失,与OVX-M小鼠相比,OVX-A-M小鼠血清SER水平下降(P<0.001)。体外实验发现,SER可促进BMSCs成骨分化,在0.1μmol/L浓度时效果最佳,且0.1μmol/L SER可提高成骨相关基因Colla1及Htr2a表达水平(P<0.05)。结论艾灸疗法可抑制绝经后骨丢失,其作用可能通过调控�Objective To investigate the regulatory effect of moxibustion,a traditional Chinese medicine therapy,on bone metabolism in ovariectomized(estrogen-deficient)mice and to explore its underlying mechanisms.Methods Female C57BL/6J mice of 12 weeks old were randomly assigned to five groups,including a sham operation control group(SHAM),an ovariectomy group(OVX),a group given ovariectomy and broad-spectrum antibiotics(OVX-A),a group given ovariectomy and moxibustion(OVX-M),and a group given ovariectomy,broad-spectrum antibiotics,and moxibustion(OVX-A-M),with 5 mice in each group.Then,4 weeks post-surgery,the mice in each group received broadspectrum antibiotics and/or moxibustion intervention for an additional 4 weeks.After that,the mice were sacrificed,and samples were collected.Micro-CT was used to assess bone volume parameters in the distal femurs,including bone volume/tissue fraction(BV/TV),trabecular number(Tb.N),trabecular thickness(Tb.Th),trabecular separation(Tb.Sp),and cortical thickness(Ct.Th).Targeted metabolomics was used to measure serum tryptophan metabolites,and qPCR was performed to quantify serotonin(SER)receptors Htr2a and Htr2b mRNA levels in bone marrow.In addition,primary bone marrow mesenchymal stem cells(BMSCs)were treated with serotonin of varying concentration gradients(0,0.01,0.1,1,and 10μmol/L).Alkaline phosphatase(ALP)staining was performed to assess osteogenic differentiation,while qPCR was performed to assess the expression of Colla1,an osteogenesis-related gene,and serotonin receptors Htr2a and Htr2b.Results Compared with the SHAM mice,the OVX mice exhibited significant deterioration in bone microarchitecture,showing decreased BV/TV([10.57±2.82]%vs.[4.20±0.96]%,P<0.01),reduced Tb.N([3.16±0.11]vs.[2.25±0.15],P<0.01),increased Tb.Sp([0.31±0.01]vs.[0.45±0.03],P<0.01),and decreased levels of serum tryptophan metabolite SER.Compared with the OVX mice,the OVX-M mice showed a notable improvement in bone microarchitecture,with BV/TV increasing to(7.51±1.42)%(P<0.05),and elevated levels of

关 键 词：绝经后骨质疏松 艾灸 肠道菌群 血清素 

分 类 号：R245.81[医药卫生—针灸推拿学]