正交设计探讨续苓健骨方对快速骨丢失模型大鼠的组方优化研究  

An orthogonal design study on the optimization of the Xuling Jiangu Formula for inducing rapid bone loss in rat models

作　　者：陈赛楠 李生强 谢丽华 陈玄 黄景文 叶云金 葛继荣 CHEN Sainan;LI Shengqiang;XIE Lihua;CHEN Xuan;HUANG Jingwen;YE Yunjin;GE Jirong(Key Research Laboratory of Osteoporosis Syndrome Genomics,Fujian Academy of Chinese Medical sciences,Fuzhou 350003;Fujian Key Laboratory of Integrated Traditional Chinese and Western Medicine for Osteoporosis(Fujian Academy of Chinese Medical Sciences,Rehabilitation Hospital Affiliated to Fujian University of TCM),Fuzhou 350003;Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122;Fujian Provincial Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou 350122,China)

机构地区：[1]福建省中医药科学院骨质疏松证候基因组学研究室,福建福州350003 [2]福建省中西医结合防治骨质疏松重点实验室(福建省中医药科学院、福建中医药大学附属康复医院),福建福州350003 [3]福建中医药大学中西医结合研究院,福建福州350122 [4]福建省中西医结合老年性疾病重点实验室,福建福州350122

出　　处：《中国骨质疏松杂志》2024年第7期1006-1012,共7页Chinese Journal of Osteoporosis

基　　金：福建省卫生健康委中医药科研课题(2021zyjc21);福建省属公益类科研院所基本科研专项(2022R1003003,2022R1003002)。

摘　　要：目的 通过正交设计观察续苓健骨方中的药物对卵巢切除术(ovariectomy, OVX)模型大鼠骨密度、骨微结构和骨代谢影响的主次作用,并探讨疗效较佳优化组别的潜在作用机制。方法 大鼠随机分为假手术组、模型组、续苓健骨方组、优化1组、优化2组、优化3组、优化4组、优化5组、优化6组、优化7组和优化8组,每组10只,采用OVX制备大鼠快速骨丢失模型,连续灌胃12周后取材备检。双能X线骨密度仪检测胫骨近端骨密度,micro-CT检测胫骨微观结构,ELISA检测血清骨吸收金指标CTX和骨形成金指标PINP的含量,Real-time PCR和Western blot检测股骨OPG、RANKL mRNA和蛋白的表达水平。结果 续苓健骨方组、优化4组、6组和7组骨密度均显著升高(P<0.05),优化4和7组Tb.N和BV/TV显著升高(P<0.001),续苓健骨方组和优化1~8组血清CTX、PINP均不同程度降低,续苓健骨方组和优化7组OPG mRNA和蛋白表达水平显著升高、RANKL mRNA和蛋白表达水平显著降低。结论 优化4组、6组和7组是治疗效果较好的组别,可通过调控OPG/RANKL发挥抗骨质疏松作用,续断、骨碎补、女贞子、红花和川芎是续苓健骨方取效的主要有效药物。Objective To investigate the primary and secondary impacts of the drug Xuling Jiangu Formula on bone mineral density(BMD),structure,and bone metabolism in ovariectomized rats experiencing rapid bone loss with an orthogonal design.The aim of this study is to elucidate the mechanism of action of the drug.Methods A total of 110 female SD rats were randomly assigned to sham surgery group,OVX group,Xuling Jiangu Formula group,and optimization groups 1-8.The model was established with bilateral ovariectomy.After a 12-week intervention of drug gavage,samples were collected to measure tibial bone density using DXA and tibial microstructure using micro-CT.ELISA was employed to detect serum levels of collagen type I cross-linked C-terminal peptide(S-CTX)and procollagen type I N-terminal precursor peptide(PINP).Real-time PCR and Western blotting were utilized to measure the mRNA and protein expression levels of OPG and RANKL in the femur.Results The result indicated a significant increase in BMD in Xuling Jiangu Formula group,optimized group 4,group 6,and group 7 compared to that in the model group(P<0.05).Tb.N and BV/TV levels in optimized groups 4 and 7 increased significantly(P<0.001).The serum levels of CTX and PINP exhibited varying degrees of reduction in both Xuling Jiangu Formula group and optimized group 1-8.Conversely,the expression levels of OPG mRNA and protein increased significantly,while the expression levels of RANKL mRNA and protein decreased significantly in Xuling Jiangu Formula group and optimized group 7.Conclusion Group 4,6,and 7 demonstrate superior therapeutic effects,suggesting the potential in exerting anti-osteoporosis effects through the regulation of OPG/RANKL.The primary effective drugs in this regard include Dipsacus,Drynaria,Llucidum,Safflower,and Chuanxiong.

关 键 词：绝经后骨质疏松症 续苓健骨方 正交设计 OPG/RANKL 中医中药 

分 类 号：R285.5[医药卫生—中药学] R-332[医药卫生—中医学]