蒙古族药忠伦阿汤对CIA大鼠MMP-3,MMP-9表达的影响

Effect of Mongolian Medicine Zhonglun'e Tang on MMP-3,MMP-9 Expressions in Rats with Collagen Induced Arthritis

目的:观察忠伦阿汤的有效成分对胶原诱导性关节炎(CIA)大鼠的基质金属蛋白酶-3(MMP-3),MMP-9的影响,探讨其治疗类风湿关节炎(RA)的作用机制。方法:采用Ⅱ型胶原诱导建立关节炎大鼠模型,并设正常组,模型组,雷公藤多苷片组(9.45 mg·kg-1),忠伦阿汤高、中、低剂量组(5.4,2.7,1.35 g·kg-1),连续给药4周,苏木素-伊红(HE)观察滑膜组织的组织病理形态,酶联免疫吸附法(ELISA)测定大鼠血清MMP-3与MMP-9的含量;免疫组化与蛋白质免疫印迹(Western blot)检测滑膜组织MMP-3与MMP-9的表达。结果:模型组血清MMP-3及MMP-9的含量显著高于正常组(P<0.01);各用药组MMP-3和MMP-9表达较模型组均有不同程度的降低(P<0.01),忠伦阿汤高剂量组与雷公藤多苷片组比较无显著差异;免疫组化结果显示MMP-3,MMP-9阳性表达模型组较正常组明显升高(P<0.01),雷公藤多苷片组及忠伦阿汤高剂量组较模型组MMP-3,MMP-9蛋白表达明显降低,有统计学意义(P<0.05),Western blot分析结果表明用药组MMP-3,MMP-9的蛋白表达明显低于模型组,差异有统计学意义(P<0.05)。结论:忠伦阿汤能明显减少CIA大鼠血清中MMP-3及MMP-9水平,降低滑膜组织的表达,可能是其治疗RA的作用机制。

Objective： To observe the effect of Mongolia medicine Zhonglun＇e Tang on the expressions of matrix metalloproteinase-3（ MMP-3） and MMP-9 in rats with collagen induced arthritis（ CIA） and to study its treatment mechanism for rheumatoid arthritis（ RA）. Method： arthritis rat models were established by using type II collagen. The rats were divided into normal group,model group,tripterygium wilfordii polycoride tablets group（ 9. 45 mg·kg- 1）,Mongolian medicine Zhonglun＇e Tang high dose（ 5. 4 g·kg- 1）,medium dose（ 2. 7 g·kg- 1）and low dose（ 1. 35 g·kg- 1） groups. The drugs were given for 4 weeks,and then HE staining was used to observe the histopathologic morphology of synovial tissues. MMP-3 and MMP-9 levels in the serum of rats were measured by enzyme linked immunosorbent assay（ ELISA）. The expressions of MMP-3 and MMP-9 in the synovial tissues were detected by using immunohistochemistry and Western blot. Result： The levels of MMP-3 and MMP-9 in serum of model group were significantly higher than those in normal group（ P〈 0. 01）; positive expressions of each drug group were lower than those in model group to varying degrees（ P〈 0. 01）,but there was no significant difference between Zhonglun＇e Tang high dose group and tripterygium wilfordii polycoride tablets group. The results of immunohistochemistry method showed that,the expressions of MMP-3 and MMP-9 in mode group were significantly higher than those in normal group（ P〈 0. 01）,and there was statistically significant difference between Zhonglun＇e Tang high dose group,tripterygium wilfordii polycoride tablets group were lower in protein expression of MMP-3and MMP-9 than those in model group（ P〈 0. 05）. The results of Western blot showed that the protein expressions of MMP-3 and MMP-9 in the treatment groups group were significantly lower than those in model group, with statistically significant difference（ P〈 0. 05）. Conclusion： Zhonglun＇e Tang could significantly decrease the MMP-3 and MMP-9 levels in serum of CIA rats,and reduce the positive expressions of synovial tissues. This may be the mechanism for treating RA.