摘要
目的:探究补肾活血方(BSHX)对实验性膝骨性关节炎兔退变软骨的保护作用及其可能的机制。方法:将新西兰兔分为对照(control)组、模型(model)组、BSHX(1.86 g/kg)组、Hedgehog通路激活剂SAG(20 mg/kg)组和BSHX+SAG组,每组6只。测定兔膝关节宽度,处死兔后观察膝关节软骨大体情况,对软骨退变情况进行评定;HE染色和番红O染色观察膝关节软骨组织病理形态学变化;ELISA法检测软骨组织中肿瘤坏死因子α(TNF-α)和白细胞介素1β(IL-1β)水平;免疫组化法检测软骨组织中基质金属蛋白酶13(MMP-13)和II型胶原(Col-II)蛋白表达;Western blot法检测软骨组织中Shh、Ptch1、Smo和Gli1蛋白表达。结果:与control组相比,model组膝关节宽度增大,软骨退变评分、软骨分级和Mankin评分升高,软骨组织中TNF-α和IL-1β水平升高,MMP-13阳性表达率升高,Col-II阳性表达率降低,Shh、Ptch1、Smo和Gli1蛋白表达升高(P<0.05)。与model组相比,BSHX组膝关节宽度减小,软骨退变评分、软骨分级和Mankin评分降低,软骨组织中TNF-α和IL-1β水平降低,MMP-13阳性表达降低,Col-II阳性表达升高,Shh、Ptch1、Smo和Gli1蛋白表达降低;SAG组膝关节宽度增大,软骨退变评分、软骨分级和Mankin评分升高,软骨组织中TNF-α和IL-1β水平升高,MMP-13阳性率表达升高,Col-II阳性表达率降低,Shh、Ptch1、Smo和Gli1蛋白表达升高(P<0.05)。BSHX+SAG组膝关节宽度,软骨退变评分,软骨分级,Mankin评分,软骨组织中TNF-α和IL-1β水平,MMP-13阳性表达率,以及Shh、Ptch1、Smo和Gli1蛋白表达均低于SAG组,Col-II阳性表达率高于SAG组(P<0.05)。结论:补肾活血方对实验性膝骨性关节炎兔退变软骨具有保护作用,其机制可能与抑制Hedgehog通路有关。
AIM:To explore the protective effect of Bushen-Huoxue prescription(BSHX)on degenerative cartilage in rabbits with experimental knee osteoarthritis and its possible mechanism.METHODS:New Zealand rabbits were divided into control group,model group,BSHX(1.86 g/kg)group,Hedgehog pathway activator SAG(20 mg/kg)group and BSHX+SAG group,with 6 rabbits in each group.The knee joint width of the rabbits was measured.After the rabbits were killed,the cartilage of knee joint was observed,and the cartilage degeneration was assessed.The histopathological changes of knee cartilage were observed by HE staining and safranine O staining.The levels of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in the cartilage tissue were measured by ELISA.Immunohistochemical method was used to detect the expression of matrix metalloproteinase-13(MMP-13)and collagen type II(Col-II)in the cartilage tissue,and the protein expression levels of Shh,Ptch1,Smo and Gli1 in the cartilage tissue were determined by Western blot.RESULTS:Compared with control group,the knee joint width,the cartilage degeneration score,the cartilage grading,the Mankin score,the TNF-αand IL-1βlevels in the cartilage tissue,and the MMP-13 positive expression rate in the cartilage tissue were increased,Col-II positive expression rate was decreased,and the protein expression of Shh,Ptch1,Smo and Gli1 was increased in model group(P<0.05).Compared with model group,the knee joint width,the cartilage degeneration score,the cartilage grading,the Mankin score,the levels of TNF-αand IL-1βin the cartilage tissue,and the MMP-13 positive expression rate in the cartilage tissue were decreased,Col-II positive expression rate was increased,and the protein expression of Shh,Ptch1,Smo and Gli1 was decreased in BSHX group(P<0.05).Compared with model group,the knee joint width,the cartilage degeneration score,the cartilage grading,the Mankin score,the levels of TNF-αand IL-1βin the cartilage tissues,and the MMP-13 positive expression rate in the cartilage tissue were increased
作者
贺自克
王上增
沈锦涛
HE Zi-ke;WANG Shang-zeng;SHEN Jin-tao(Department of Arthrology,Henan Hospital of Traditional Chinese Medicine,Zhengzhou 450000,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2020年第4期700-706,共7页
Chinese Journal of Pathophysiology
基金
河南省中医药科学研究专项课题(No.2019ZY2059)。
