基于“肠-骨”轴探讨药食同源复方强骨宁缓解绝经后骨质疏松症的作用机制  

Exploration on the mechanism of the combination of medicine and food homogeneous compound Qianggu Ning in the remission of postmenopausal osteoporosis based on the‘gut-bone'axis

作　　者：李宁 吴月滢 裴环 李律宇 任煜 李蕾 王斯 袁嘉丽 LI Ning;WU Yueying;PEI Huan;LI Lvyu;REN Yu;LI Lei;WANG Si;YUAN Jjiali(The First Clinical Medical College of Yunnan University of Chinese Medicine,Kunming 650500,China;Yunnan Provincial Key Laboratory of Integrated Traditional Chinese and Western Medicine for Chronic Disease in Prevention and Treatment,Yunnan University of Chinese Medicine,Kunming 650500,China;Kunming Municipal Hospital of Traditional Chinese Medicine,Third Affiliated Hospital of Yunnan University of Chinese Medicine,Kunming650500,China)

机构地区：[1]云南中医药大学第一临床医学院,昆明650500 [2]云南中医药大学云南省中西医结合慢病防治重点实验室,昆明650500 [3]昆明市中医医院,云南中医药大学第三附属医院,昆明650500

出　　处：《中华中医药杂志》2024年第5期2308-2315,共8页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：云南省科技计划项目-重大科技专项计划-生物种业和农产品精深加工专项(No.202102AE090031);云南省教育厅科学研究基金项目(No.2022Y354);云南省科技厅科技人才与平台计划-云南省中医药学分子生物学重点实验室(No.2019DG016);云南省科技厅-中医联合专项面上项目(No.202101AG070053)。

摘　　要：目的:从“肠-骨”轴角度探索强骨宁缓解绝经后骨质疏松症(PMOP)的作用机制。方法:采用双侧卵巢去势(OVX)法模拟PMOP模型,将雌性C57BL/6J小鼠随机分为假手术组(Sham组),卵巢去势组(OVX组),强骨宁低、中、高剂量组,连续灌胃给药8周后收集小鼠股骨、结肠、新鲜粪便。Micro-CT检测小鼠股骨骨密度变化,Masson染色观察股骨骨微结构与胶原纤维变化,ELISA检测血清骨转换因子和结肠中炎症因子的表达水平,免疫荧光检测结肠中Claudin-1、ZO-1、Occludin表达,16S rDNA高通量测序检测分析粪便中微生物的物种变化,CG-MS检测粪便中短链脂肪酸(SCFAs)的含量。结果:与Sham组比较,OVX组小鼠股骨骨密度显著降低,骨小梁稀疏、断裂,骨转换率提高;肠道炎症水平升高,肠屏障损伤,肠道菌群多样性呈减少趋势,粪便中SCFAs含量减少。与OVX组比较,强骨宁干预后OVX小鼠骨密度及股骨骨微结构明显改善,骨转换率降低;肠道炎症水平下降,肠道紧密连接蛋白表达水平恢复,肠道菌群中g_Akkermansia、g_Parabacteroides、g_Alistipes等优势菌属丰度显著增加(P<0.05),g_Ruminococcus菌属丰度显著减少(P<0.05),SCFAs表达量增加。结论:强骨宁可缓解OVX小鼠的骨丢失及其伴随的肠道炎症及屏障损伤,其作用机制可能与调控肠道菌群与SCFAs的变化密切相关。Objective:To explore the mechanism of Qianggu Ning(QGN)in relieving postmenopausal osteoporosis(PMOP)from the perspective of the‘gut-bone'axis.Methods:The bilateral ovariectomized(OVX)method was used to simulate a PMOP model.Female C57BL/6J mice were randomly divided into sham surgery group(Sham group),ovariectomized group(OVX group),and QGN low,medium,and high dose group(QGN-L,QGN-M,QGN-G).After continuous gavage for 8 weeks,the femur,colon,and fresh feces of the mice were collected.Micro-CT was used to detect changes in femoral bone density in mice,Masson staining was used to observe changes in femoral bone microstructure and collagen fibers,ELISA was used to detect the expression levels of serum bone turnover factors and inflammatory factors in the colon,immunofluorescence was used to detect the expression of Claudin-1,ZO-1,and Occludin in the colon,16S rDNA high-throughput sequencing was used to analyze changes in microbial species in feces,and CG-MS was used to detect the content of SCFAs in feces.Results:Compared with the Sham group,the OVX group mice showed a significant decrease in femoral bone density,sparse and fractured bone trabeculae,and an increase in bone turnover rate.The level of intestinal inflammation increases,the intestinal barrier is damaged,the diversity of intestinal microbiota shows a decreasing trend,and the content of SCFAs in feces decreases.Compared with the OVX group,QGN intervention significantly improved bone density and femoral bone microstructure in OVX mice,and reduced bone turnover rate.The level of intestinal inflammation decreased,the expression level of intestinal tight junction protein recovered,and the abundance of dominant bacterial genera such as g_Akkermansia,g_Paraacteroides,and g_Alistipes in the gut microbiota significantly increased(P<0.05),while the abundance of g_Ruminococcus bacteria significantly decreased(P<0.05),and the expression level of SCFAs increased.Conclusion:QGN can alleviate bone loss and associated intestinal inflammation and barrier damage in OVX

关 键 词：“肠-骨”轴 强骨宁 肠屏障 16S rDNA 短链脂肪酸 绝经后骨质疏松症 

分 类 号：R285.5[医药卫生—中药学]