消肿止痛合剂对大鼠膝骨性关节炎软骨中PI3K/Akt信号通路影响的实验研究

Experimental study on the effect of Xiao Zhong Zhi Tong mixture on PI3K/Akt signaling pathway in cartilage of knee osteoarthritis in rats

目的观察消肿止痛合剂对大鼠膝骨性关节炎软骨中磷脂酰肌醇3激酶(PI3K)蛋白和丝氨酸苏氨酸蛋白激酶(AKT)蛋白、天冬氨酸蛋白水解酶3(Caspase-3)表达的影响及其作用机制。方法将80只SD大鼠随机分为4组:观察组(消肿止痛合剂组)、对照组(硫酸氨基葡萄糖片组)、模型组、假手术组,每组20只。除假手术外,其余三组通过改良Hulth法行膝骨关节炎造模。药物连续干预8周后,取各组大鼠膝关节软骨,采用ELISA方法检测血清中TNF-α、IL-1β、iNOS水平,实时荧光定量聚合酶链式反应(Real-time PCR)检测各组软骨中PI3K、AKT、Caspase-3mRNA的表达情况,蛋白质印迹法(Western blot)检测各组软骨细胞中PI3K、AKT、Caspase-3蛋白的表达。结果消肿止痛合剂可显著减低KOA大鼠血清TNF-α、IL-1β、iNOS水平,显著下调PI3K、AKT、Caspase-3基因及蛋白表达。结论消肿止痛合剂对KOA关节软骨损伤的保护可能与PI3K/AKT信号通路相关。

Objective To observe the effect of elimination of swelling and analgesic mixture on the expression of phosphatidylinositol 3 kinase(PI3K)protein and serine protein kinase(AKT)protein and aspartate protein hydrolase 3(Caspase-3)in the cartilage of rats with osteoarthritis of the knee.Methods 80 SD rats were randomly divided into 4 groups:the observation group(anti swelling and analgesic mixture group),the control group(Glucosamine Sulfate Tablets group),the model group and the sham operation group(20 rats in each group),except for the false operation,the other 3 groups were made by modified Hulth method for knee osteoarthritis.After 8 weeks of continuous intervention,the cartilage of the knee joint was taken and the levels of TNF-alpha,IL-1 beta and iNOS were detected by ELISA method.The expression of PI3K,AKT and Caspase-3mRNA in cartilage of each group was detected by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR),and Western blot(Western blot)was used to detect the soft tissue of each group.The expression of PI3K,AKT and Caspase-3 protein in bone cells.Results the swelling swelling and pain relieving mixture can significantly reduce the levels of serum TNF-alpha,IL-1 beta and iNOS in KOA rats,and significantly reduce the expression of PI3K,AKT and Caspase-3 genes and protein.Conclusion the protective effect of Xiao Zhong Zhi Tong mixture on KOA articular cartilage injury may be related to PI3K/AKT signaling pathway.