右归丸对膝骨关节炎模型鼠软骨组织显微结构及热休克蛋白90α蛋白表达的影响

目的:观察右归丸对膝骨关节炎(knee osteoarthritis,KOA)模型大鼠软骨组织显微结构和热休克蛋白90α(heat shock protein 90α,Hsp90α)蛋白表达的影响,进一步揭示右归丸防治KOA的机制。方法:将大鼠随机分为假手术对照组、KOA模型组、硫酸氨基葡萄糖组和右归丸(高、中、低剂量)组,每组10只。采用改良Hulth法制备大鼠KOA模型,分别予相应药物灌胃8周。HE染色法观察软骨组织的形态学变化,并进行Makin评分;应用免疫组化法检测各组大鼠软骨组织Hsp90α、纤连蛋白(fibronectin,Fn)和Ⅱ型胶原(collagen typeⅡ,COL-Ⅱ)的表达;RT-qPCR法检测各组大鼠软骨组织Hsp90α、白细胞介素1β(interleukin-1β,IL-1β)、基质金属蛋白酶3(matrix metalloproteinase-3,MMP-3)和MMP-13的mRNA表达水平,应用Western blot法检测各组大鼠软骨组织Hsp90α和COL-Ⅱ的表达。结果:与假手术组比较,KOA模型组大鼠软骨组织Makin评分明显升高,软骨组织H...

Effects of Youguiwan on microstructure and Hsp90 of chondrocytes in rats with knee osteoarthritis

AIM: To observe the effect of Youguiwan(YGW) on the expression of heat shock protein 90α(Hsp90α) in articular cartilage tissues of knee osteoarthritis(KOA) rats, and to further reveal its mechanism. ME-THODS: SD rats(n=60) were randomly divided into 6 groups: sham control(SC) group, model(M) group, glucosamine sulfate(GS) group, and high-, middle-and low-dose YGW(YGW-H, YGW-M and YGW-L) groups. The modified Hulth method was used to make KOA models for 6 weeks. The rats were gavaged with corresponding drugs for 8 weeks. HE staining was used and Makin score was evaluated. The expression of Hsp90α, fibronectin(Fn) and collagen type Ⅱ(COL-II) was determined by immunohistochemistry. The expression of interleukin-1β(IL-1β), matrix metalloproteinase(MMP)-3 and MMP-13 was analyzed by RT-qPCR. The protein expression of Hsp90α and COL-II was determined by Western blot. RESULTS: As compared with SC group, the Makin score was obviously raised in M group, the expression levels of Hsp90α, IL-1β, MMP-3 and MMP-13 were evidently increased, the expression levels of COL-II and Fn were evidently decreased(P<0.01), articular cartilage was seriously damaged, and the chondrocytes were disarranged. As compared with M group, the Makin score and the expression of Hsp90α were obviously decreased in YGW-H group, the protein expression of Fn was evidently increased in YGW-H group, the protein expression of COL-II was evidently increased in YGW-H and YGW-M groups, the mRNA expressions of IL-1β, MMP-3 and MMP-13 were evidently decreased in YGW-H, YGW-M and YGW-L groups(P<0.05 or P<0.01), cartilage structure tended to be normal, the chondrocytes distribution was uneven, and articular cartilage surface was not smooth. CONCLUSION: Youguiwan is effective for treatment of KOA. Youguiwan protects against articular cartilage degeneration by down-regulating the expression of Hsp90α and up-regulating the expression of Fn, and reducing the secretion of inflammatory factors and the degradation of extracellular matrix.