肾髓安康饮干预小鼠膝骨关节炎的效果和安全性及作用机制研究  

Efficacy,safety,and mechanism of Shensui Ankang Yin(肾髓安康饮)against knee osteoarthritis in mice:an experimental study

作　　者：李雪枫 王玉杭 邹凯奥 温经渊 陈文哲 袁文华 肖鲁伟[2] 金红婷[2] LI Xuefeng;WANG Yuhang;ZOU Kaiao;WEN Jingyuan;CHEN Wenzhe;YUAN Wenhua;XIAO Luwei;JIN Hongting(The First Clinical Medical College of Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang,China;Zhejiang Provincial Hospital of Traditional Chinese Medicine,Hangzhou 310006,Zhejiang,China)

机构地区：[1]浙江中医药大学第一临床医学院,浙江杭州310053 [2]浙江省中医院,浙江杭州310006

出　　处：《中医正骨》2025年第2期4-12,共9页The Journal of Traditional Chinese Orthopedics and Traumatology

基　　金：国家自然科学基金项目(82274280);浙江省基础公益研究计划项目(LR23H270001)。

摘　　要：目的:探讨肾髓安康饮干预小鼠膝骨关节炎(knee osteoarthritis,KOA)的效果、安全性及作用机制。方法:将30只C57BL/6J小鼠随机分为假手术组、模型组和肾髓安康饮组。模型组和肾髓安康饮组小鼠采用内侧半月板失稳术在右后肢构建KOA模型,假手术组小鼠切开右后肢皮肤、暴露膝关节后缝合。自造模手术后第2天起,肾髓安康饮组小鼠以肾髓安康饮浓缩液(生药浓度1.6 g·mL^(-1))灌胃,每次0.5 mL,每天1次,连续灌胃8周;模型组和假手术组小鼠以等量生理盐水灌胃。干预8周后,分别采用Up-down法和热板实验测定小鼠机械痛阈值和热痛阈值,采用DigiGait步态分析系统分析小鼠步态;采用Micro-CT扫描仪扫描小鼠右膝关节,测定胫骨平台软骨下骨的骨体积分数;制备小鼠膝关节软骨组织切片,采用阿尔辛蓝-苏木素/橙黄G染色观察膝关节软骨退变情况,采用国际骨关节炎研究学会(Osteoarthritis Research Society International,OARSI)评分标准评价膝关节软骨退变情况,采用免疫组织化学染色法检测膝关节软骨中Ⅱ型胶原、基质金属蛋白酶(matrix metalloproteinase,MMP)-13的表达情况;制备小鼠膝关节软骨下骨和L_(4)右侧背根神经节组织切片,采用免疫荧光染色法检测小鼠膝关节软骨下骨和L_(4)右侧背根神经节内神经生长因子(nerve growth factor,NGF)的表达情况;取小鼠心脏、肝脏、脾脏、肺脏、肾脏,制备组织切片,采用HE染色法观察小鼠内脏组织病变情况。结果:①疼痛阈值测定结果。模型组小鼠50%缩足阈值低于假手术组(P=0.000),右后足反应时间短于假手术组(P=0.000);肾髓安康饮组小鼠50%缩足阈值高于模型组(P=0.000),右后足反应时间长于模型组(P=0.000)。②步态分析结果。模型组小鼠右后肢的摆动相时间长于假手术组(P=0.001),支撑相时间短于假手术组(P=0.000),步幅和足底接触面积均小于假手术组(P=0.000,P=0.000);肾髓安康�Objective:To observe the outcomes and safety of Shensui Ankang Yin(肾髓安康饮,SSAKY)against knee osteoarthritis(KOA)in mice,and to explore its mechanism.Methods:Thirty C57BL/6J mice were selected and randomized into sham-operated group,model group,and SSAKY group.The mice in model group and SSAKY group were subjected to destabilization of the medial meniscus(DMM)on the right hind limbs to build KOA models,while the ones in sham-operated group were merely incised the skin at the corresponding site and then sutured after exposing the knee joints.From day 2 after the modeling surgery,the mice in SSAKY group were intervened by intragastric administration with SSAKY concentrate(the crude drug concentration was 1.6 g/mL),once a day,0.5 mL at a time for consecutive 8 weeks;while the ones in model group and sham-operated group with the same dosage of normal saline.After 8-week intervention,the mechanical and thermal pain thresholds of the mice were measured by Up-down method and hot plate test,respectively,and the gait of the mice was analyzed using the DigiGait gait analysis system.After that,the mice were executed,and the knee joints were harvested from their right hind limbs,and scanned by Micro-CT scanner to measure the bone volume fraction(BVF)of the subchondral bone in tibial plateau.Furthermore,the knee joint cartilage tissue sections were prepared and stained with alcian blue-hematoxylin/orange G(ABH/OG)to observe the degeneration of knee joint cartilage,and then evaluate by using Osteoarthritis Research Society International(OARSI)scoring criteria,apart from that,the expression of typeⅡcollagen and matrix metalloproteinase(MMP)-13 in the knee joint cartilages was detected by employing immunohistochemical staining.In addition,the tissue sections of the subchondral bone of mice knee joints and the right dorsal root ganglion(DRG)of L_(4) were prepared for detecting the expression of nerve growth factor(NGF)by immunofluorescence staining;besides,the heart,liver,spleen,lung and kidney of mice were harvested to

关 键 词：骨关节炎 膝 肾髓安康饮 疼痛 敏化 软骨细胞 神经生长因子 小鼠 

分 类 号：R285.5[医药卫生—中药学]