苗药验方对膝骨关节炎家兔血清β-catenin、TNF-α水平的影响

Effect of Miao medicine prescription on the levels ofβ-catenin and TNF-αin rabbits with knee osteoarthritis

目的通过观察苗药验方对膝骨关节炎家兔血清β-连环蛋白(β-catenin)、肿瘤坏死因子-α(TNF-α)的影响,探讨苗药验方防治膝骨关节炎软骨退变的作用机制。方法36只家兔,随机分为正常对照组、模型组、苗药熏蒸组、苗药离子导入组,每组9只。除正常对照组外,其余三组家兔右膝关节腔内注射木瓜蛋白酶制造膝骨关节炎模型,造模成功后,苗药熏蒸组与苗药离子导入组家兔分别经苗药熏蒸治疗及苗药离子导入治疗20 d,于治疗后第10天及第20天进行膝关节改良Lequesne MG评分,第21天采集四组家兔血液样本,运用酶联免疫吸附测定(ELISA)检测法检测四组家免血清中β-catenin、TNF-α水平。结果正常对照组、模型组、苗药熏蒸组、苗药离子导入组家兔第10天的改良Lequesne MG评分分别为(0.22±0.44)、(6.78±0.97)、(4.56±1.67)、(3.56±1.01)分,第20天的改良Lequesne MG评分分别为(0.33±0.71)、(6.11±0.78)、(2.67±0.71)、(2.44±0.88)分。模型组、苗药熏蒸组、苗药离子导入组家兔第10、20天的改良Lequesne MG评分均高于正常对照组,差异有统计学意义(P<0.01);苗药熏蒸组、苗药离子导入组家兔第10、20天的改良Lequesne MG评分均低于模型组,差异有统计学意义(P<0.01);苗药熏蒸组、苗药离子导入组家兔第20天的改良Lequesne MG评分均低于本组第10天,差异有统计学意义(P<0.05)。正常对照组、模型组、苗药熏蒸组、苗药离子导入组家兔血清β-catenin水平分别为(46.05±10.50)、(83.06±4.76)、(68.12±10.70)、(69.63±5.68)pg/ml,血清TNF-α水平分别为(34.78±10.23)、(64.28±8.44)、(49.20±7.14)、(57.14±6.00)pg/ml。模型组、苗药熏蒸组、苗药离子导入组家兔血清β-catenin、TNF-α水平均高于正常对照组,差异有统计学意义(P<0.01);苗药熏蒸组家兔血清β-catenin、TNF-α水平均低于模型组,苗药离子导入组家兔血清β-catenin水平低于模型组,差异有统计学意义(P<0.01);苗药离子导入组家兔血清TNF-α水平高于苗药熏蒸组,差异具有统计学意义(P<0.05)。结论苗药验方可能通过下调血清β-catenin、TNF-α的水平,抑制炎症反应,改善症状,延缓膝骨关节炎的进展。

Objective To observe the effect of Miao medicine proven prescription on the levels ofβ-catenin and tumor necrosis factor-α(TNF-α)in rabbits with knee osteoarthritis,so as to discuss the mechanism of Miao medicine prescription in preventing cartilage degeneration of osteoarthritis.Methods A total of 36 rabbits were randomly divided into normal control group,model group,Miao medicine fumigation group,Miao medicine iontophoresis group,with 9 rabbits in each group.In addition to the normal control group,the remaining three groups were injected papain into the right knee joint cavity to make knee osteoarthritis model.After successful modeling,the rabbits in the Miao medicine fumigation group and Miao medicine iontophoresis group were treated with Miao medicine fumigation and Miao medicine iontophoresis for 20 d.Modified Lequesne MG score of the knee joint was performed on the 10th and 20th day after treatment.The blood samples from each group of rabbits on the 21st day were collected,and the enzyme-linked immunosorbent assay(ELISA)was used to detect the levels ofβ-catenin and TNF-αin the serum of rabbits in each group.Results The modified Lequesne MG scores of the rabbits in the normal control group,model group,Miao medicine fumigation group and Miao medicine iontophoresis group on the 10th day were(0.22±0.44),(6.78±0.97),(4.56±1.67),and(3.56±1.01)points,and the modified Lequesne MG scores on the 20th day were(0.33±0.71),(6.11±0.78),(2.67±0.71),and(2.44±0.88)points.On the 10th and 20th day,the modified Lequesne MG scores of the rabbits in the model group,Miao medicine fumigation group and Miao medicine iontophoresis group were higher than those in the normal control group,and the differences were statistically significant(P<0.01).On the 10th and 20th day,the modified Lequesne MG scores of the rabbits in the Miao medicine fumigation group and Miao medicine iontophoresis group were lower than those in the model group,and the differences were statistically significant(P<0.01).The modified Lequesne MG score of the rabbits in the Miao medicine fumigation group and Miao medicine iontophoresis group on the 20th day was lower than that on the 10th day in this group,and the difference was statistically significant(P<0.05).The serumβ-catenin levels in the normal control group,model group,Miao medicine fumigation group and Miao medicine iontophoresis group were(46.05±10.50),(83.06±4.76),(68.12±10.70)and(69.63±5.68)pg/ml,and the serum TNF-αlevels were(34.78±10.23),(64.28±8.44),(49.20±7.14)and(57.14±6.00)pg/ml.The serum levels ofβ-catenin and TNF-αin the model group,Miao medicine fumigation group and Miao medicine iontophoresis group were higher than those in the normal control group,and the differences were statistically significant(P<0.01).The serumβ-catenin and TNF-αlevels of the rabbits in the Miao medicine fumigation group were lower than those in the model group,and the serumβ-catenin level in the rabbits in the Miao medicine iontophoresis group were lower than that in the model group,and the differences were statistically significant(P<0.01).The serum TNF-αlevel of the rabbits in the Miao medicine iontophoresis group was higher than that in the Miao medicine fumigation group,and the difference was statistically significant(P<0.05).Conclusion Miao medicine prescription may inhibit the inflammatory response,improve symptoms and delay the progression of knee osteoarthritis by down-regulating serumβ-catenin and TNF-αlevels.