Yan Chun-Lu, Li Sheng-Hua, An Fang-Yu, Liu Yong-Qi, Xia Peng-Fei, Ma Zheng-Min, Niu Yan-Qiang, Liu Peng-Yao
Teaching Experiment Training Center, Gansu University of Traditional Chinese Medicine, Lanzhou 730000, China.
Provincial-Level Key Laboratory for Chemistry and Quality Control of Chinese Tibetan Medicine, Lanzhou 730000, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2018 Feb 8;34(2):177-181. doi: 10.12047/j.cjap.5603.2018.043.
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.
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 (=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.
Compared with the sham group, the articular cartilage was severely damaged, the Mankin score was increased significantly (<0. 05), the mRNA and protein expression levels of DKK1 in cartilaginous tissue were markedly decreased(<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were increased significantly in model group(<0.05). Compared with model group, the articular cartilage lesions was light (<0.05), the Mankin Score was decreased significantly(<0.05), and the mRNA and protein levels of DKK1 in cartilaginous tissue were increased(<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were decreased in Yougui pill high-dose group and glucosamine sulfate group (<0.05).
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.
观察右归丸（中药）对膝骨关节炎（KOA）大鼠Wnt信号通路相关因子的影响，探讨其保护机制。
将60只SPF级SD大鼠随机分为假手术组、模型组、硫酸氨基葡萄糖组、右归丸高剂量、中剂量、低剂量治疗组（每组10只）。采用改良Hulth法建立KOA模型，持续6周。右归丸高、中、低剂量组大鼠分别按20、10、5 g/kg的剂量每日灌胃给药1次，共8周，假手术组和模型对照组给予等体积生理盐水，硫酸氨基葡萄糖组给予等体积硫酸氨基葡萄糖（1.7 g/kg·d），持续8周。末次给药后取出膝关节，显微镜下观察软骨组织病理变化。采用RT-PCR法分析大鼠软骨组织中Dickkopf同源物1（DKK1）、Wnt诱导分泌蛋白1（WISP1）、Wnt1、低密度脂蛋白受体相关蛋白5（LRP5）和β-连环蛋白的mRNA水平，采用Western blot法检测软骨组织中DKK1、WISP1、Wnt1、LRP5和β-连环蛋白的蛋白含量。
与假手术组相比，模型组关节软骨严重损伤，Mankin评分显著升高（P<0.05），软骨组织中DKK1的mRNA和蛋白表达水平显著降低（P<0.05），而WISP、Wnt1、LRP5和β-连环蛋白的表达显著升高（P<0.05）。与模型组相比，右归丸高剂量组和硫酸氨基葡萄糖组关节软骨损伤较轻（P<0.05），Mankin评分显著降低（P<0.05），软骨组织中DKK1的mRNA和蛋白水平升高（P<0.05），而WISP、Wnt1、LRP5和β-连环蛋白的表达降低（P<0.05）。
右归丸通过抑制Wnt信号通路中WISP、Wnt1、LRP5、β-连环蛋白的表达，增加DKK1细胞因子的表达，对KOA具有保护作用。
