摘要
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Hsp90α的蛋白表达显著升高,IL-1β、MMP-3和MMP-13的mRNA表达明显升高,COL-Ⅱ和Fn的蛋白表达明显降低(P<0.01),关节软骨边缘严重破坏,软骨细胞排列紊乱。与KOA模型组相比,右归丸高剂量干预组大鼠软骨组织Makin评分和Hsp90α的蛋白表达均明显降低,Fn的蛋白表达明显升高,右归丸中、高剂量组COL-Ⅱ的蛋白表达明显升高,右归丸各干预组IL-1β、MMP-3和MMP-13的mRNA表达均明显降低(P<0.05或P<0.01),软骨结构趋于正常,软骨细胞分布仅偶见不均,关节软骨表面欠光滑。结论:右归丸通过下调Hsp90α的蛋白表达并上调Fn的蛋白表达来减缓炎症因子的分泌和细胞外基质的降解,从而有效保护KOA模型大鼠关节软骨,延缓关节软骨退变。
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.
作者
安方玉
颜春鲁
刘永琦
王继龙
赵磊
夏鹏飞
骆亚莉
王国文
赵崇博
邓婕
AN Fang-yu;YAN Chun-lu;LIU Yong-qi;WANG Ji-long;ZHAO Lei;XIA Peng-fei;LOU Ya-li;WANG Guo-wen;ZHAO Chong-bo;DENG Jie(Gansu University of Traditional Chinese Medicine,Lanzhou 730000,China;Provincial Key Laboratory for Molecular Medicine of Major Diseases and the Study on Prevention and Treatment of Traditional Chinese Medicine,Gansu University of Traditional Chinese Medicine,Lanzhou 730000,China;Key Laboratory of Medicinal Chemistry and Quality Research,Colleges of Gansu Province,Lanzhou 730000,China;Key Laboratory of Dunhuang Medicine,Ministry of Education,Lanzhou 730000,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2020年第1期157-163,共7页
Chinese Journal of Pathophysiology
基金
甘肃省高等学校科研项目(No.2018A-043)
甘肃省中医药管理局科研项目(No.GZK-2017-2)
敦煌医学与转化省部共建教育部重点实验室开放基金资助项目(No.DHYX17-10)
甘肃省高校中(藏)药化学与质量研究省级重点实验室开放基金资助项目(No.zzy-2018-01)
甘肃省高校重大疾病分子医学与中医药防治研究重点实验室开放基金资助项目(No.FZYX17-18-11)
甘肃省高校大学生就业创业能力提升工程项目(No.6-1)
