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右归丸对鼠膝骨性关节炎的治疗作用及Wnt信号通路相关因子表达的影响
引用本文:颜春鲁,李盛华,安方玉,刘永琦,夏鹏飞,马正民,牛彦强,柳鹏瑶.右归丸对鼠膝骨性关节炎的治疗作用及Wnt信号通路相关因子表达的影响[J].中国应用生理学杂志,2018,34(2):177-181.
作者姓名:颜春鲁  李盛华  安方玉  刘永琦  夏鹏飞  马正民  牛彦强  柳鹏瑶
作者单位:1. 甘肃中医药大学教学实验实训中心, 兰州 730000; 2. 微生物与免疫教研室, 兰州 730000; 3. 科研实验中心, 兰州 730000; 4. 中医临床学院, 兰州 730000; 5. 临床学院, 兰州 730000; 6. 甘肃省高校中藏药化学与质量研究省级重点实验室, 兰州 730000
基金项目:甘肃省中医药管理局科研项目(GZK-2017-2);兰州市城关区科技局项目(2016-7-16);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项目(zzy-2015-04);甘肃中医药大学教学研究与教学改革项目(ZH-201606)
摘    要:目的:分析右归丸对膝骨性关节炎(KOA)大鼠Wnt信号通路相关因子表达的影响,探讨右归丸对KOA大鼠的保护机制。方法:SPF级SD大鼠60只,按照体重法随机分为假手术组、模型组、硫酸氨基葡萄糖组、右归丸高、中、低剂量组(n=10)。采用改良Hulth法复制膝骨关节炎大鼠模型。右归丸高中低剂量组分别按20、10、5 g/kg灌服相应的药物,硫酸氨基葡萄糖组按0.17 g/kg灌服硫酸氨基葡萄糖,假手术组和模型组灌服等体积的生理盐水,干预8周。末次给药后摘取膝关节,通过膝关节病理切片观察各组大鼠软骨组织病理改变;采用RT-PCR法对各组大鼠软骨组织DKK1、WISP1、Wnt1、β-catenin和LRP5 mRNA的表达水平进行对比分析;通过Western blot法检测各组大鼠软骨组织DKK1、WISP1、Wnt1、LRP5和β-catenin的蛋白质含量的变化。结果:与假手术组比较,模型组大鼠关节软骨受损严重,Mankin评分明显升高(P<0.05);DKK1 mRNA表达水平和蛋白质表达水平明显降低(P<0.05);WISP1、Wnt1、β-catenin、LRP5 mRNA表达水平及蛋白质表达水平明显升高(P<0.05)。与模型组比较,右归丸高剂量组和硫酸氨基葡萄糖组关节软骨病变明显减轻;Mankin评分明显减轻(P<0.05);大鼠软骨组织中DKK1 mRNA表达水平和蛋白表达水平明显升高,WISP1、Wnt1、β-catenin、LRP5 mRNA及蛋白表达水平显著降低(P<0.05)。结论:右归丸通过抑制Wnt信号通路中WISP1、Wnt1、β-catenin、LRP5的表达,促进DKK1细胞因子的表达,发挥对KOA的保护作用。

关 键 词:右归丸  膝骨性关节炎  Wnt信号通路  保护作用  大鼠  
收稿时间:2017-05-24

The therapeutic effects of Yougui pill on knee osteoarthritis and the expression of Wnt signal pathway related factors in rats
YAN Chun-lu,LI Sheng-hua,AN Fang-yu,LIU Yong-qi,XIA Peng-fei,MA Zheng-min,NIU Yan-qiang,LIU Peng-yao.The therapeutic effects of Yougui pill on knee osteoarthritis and the expression of Wnt signal pathway related factors in rats[J].Chinese Journal of Applied Physiology,2018,34(2):177-181.
Authors:YAN Chun-lu  LI Sheng-hua  AN Fang-yu  LIU Yong-qi  XIA Peng-fei  MA Zheng-min  NIU Yan-qiang  LIU Peng-yao
Affiliation:1. Teaching Experiment Training Center, Gansu University of Traditional Chinese Medicine, Lanzhou 730000, China; 2. Department of Microbiology and Immunology, Lanzhou 730000, China; 3. Scientific Research Experiment Center, Lanzhou 730000, China; 4. Clinical College of traditional Chinese Medicine, Lanzhou 730000, China; 5. Clinical College, Lanzhou 730000, China; 6. Provincial-Level Key Laboratory for Chemistry and Quality Control of Chinese Tibetan Medicine, Lanzhou 730000, China
Abstract:Objective:To observe the effects of Yougui pill (Traditional Chinese Medicine) on the related factors of Wnt signal pathway of rats with knee osteoarthritis (KOA), and explore its protective mechanism. Methods:Sixty SPF SD rats were randomly divided into the sham-operative group, model group, glucosamine sulfate group, high-dose, middle-dose, low-dose of Yougui pill treated group (n=10). KOA model was established by modified Hulth method for six weeks. The rats in the high, middle and low-dose of Yougui pill group were treated with Yougui pills at the doses of 20,10 and 5 g/kg respectively by gastrogavage once a day for 8 weeks, while equal volume of normal saline was given to those in the sham and model control group and an equal volume of glucosamine sulfate (1.7 g/kg·d) was given to those in glucosamine sulfate group for 8 weeks. The knee joint was removed after the last dose of drug. The pathological changes of cartilaginous tissues were observed under a microscope. The mRNA levels of Dickkopf homolog 1(DKK1), Wnt induced secreted protein 1(WISP1), Wnt1, low density lipoprotein receptor related protein 5(LRP5) and beta -catenin in rats cartilaginous tissues were analyzed by using RT-PCR method, and the protein contents of DKK1, WISP1, Wnt1, LRP5 and beta-catenin in cartilaginous tissues were detected by Western blot. Results:Compared with the sham group, the articular cartilage was severely damaged, the Mankin score was increased significantly (P<0. 05), the mRNA and protein expression levels of DKK1 in cartilaginous tissue were markedly decreased(P<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were increased significantly in model group(P<0.05). Compared with model group, the articular cartilage lesions was light (P<0.05), the Mankin Score was decreased significantly(P<0.05), and the mRNA and protein levels of DKK1 in cartilaginous tissue were increased(P<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were decreased in Yougui pill high-dose group and glucosamine sulfate group (P<0.05). Conclusion:Yougui pill has protective effects on the KOA by inhibiting the expressions of WISP, Wnt1, LRP5, beta-catenin and increasing the expression of DKK1 cytokine in the Wnt signaling pathway.
Keywords:Yougui pill  osteoarthritis of knee  Wnt signal pathway  protection funtion  rats  
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