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
陈赛楠 1李生强 2谢丽华 2陈玄 2黄景文 2叶云金 2葛继荣2
作者信息
- 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;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;福建中医药大学中西医结合研究院,福建 福州 350122;福建省中西医结合老年性疾病重点实验室,福建 福州 350122
- 2. 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;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
- 折叠
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.
Abstract
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Key words
postmenopausal osteoporosis/Xuling Jiangu Formula/orthogonal design/OPG/RANKL/traditional Chinese medicine引用本文复制引用
基金项目
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)
福建省属公益类科研院所基本科研专项(2022R1003002)
出版年
2024
NETL
NSTL
万方数据