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中国应用生理学杂志 ›› 2018, Vol. 34 ›› Issue (2): 177-181.doi: 10.12047/j.cjap.5603.2018.043

• 研究论文 • 上一篇    下一篇

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

颜春鲁1,6, 李盛华4, 安方玉1, 刘永琦2, 夏鹏飞3, 马正民4, 牛彦强5, 柳鹏瑶4   

  1. 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
  • 收稿日期:2017-05-24 修回日期:2017-10-31 出版日期:2018-03-28 发布日期:2018-05-22
  • 通讯作者: 李盛华 E-mail:yanchl1979@126.com
  • 基金资助:
    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)

The therapeutic effects of Yougui pill on knee osteoarthritis and the expression of Wnt signal pathway related factors in rats

YAN Chun-lu1,6, LI Sheng-hua4, AN Fang-yu1, LIU Yong-qi2, XIA Peng-fei3, MA Zheng-min4, NIU Yan-qiang5, LIU Peng-yao4   

  1. 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
  • Received:2017-05-24 Revised:2017-10-31 Online:2018-03-28 Published:2018-05-22
  • Supported by:
    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

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.

Key words: Yougui pill, osteoarthritis of knee, Wnt signal pathway, protection funtion, rats

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