森登-4汤对修复膝关节骨性关节炎家兔的软骨胶原纤维结构平衡的作用机制研究

Effect and mechanism of Sundon-4 Decoction on the repair cartilage collagen fiber structure balance in knee osteoarthritis rabbits

目的探讨森登-4汤对膝关节骨性关节炎(KOA)家兔的软骨组织Ⅱ型胶原蛋白(ColⅡ)、基质金属蛋白酶13(MMP-13)、MMP-9和基质金属蛋白酶组织抑制剂3(TIMP-3)的影响。方法用改良Hulth法建立家兔KOA模型。按照体重将家兔随机分为4组:模型组、实验组、阳性对照组和假手术组,每组8只。模型建立后,阳性对照组灌胃壮骨关节丸混悬液0.51 g·kg^-1,实验组灌胃蒙药森登-4汤水煎液0.43 g·kg^-1。给药4周后,取关节软骨组织,以免疫组织化学法检测ColⅡ表达(光密度比值);蛋白质免疫印迹法检测MMP-9、MMP-13和TIMP-3蛋白表达(灰度值比值)。结果假手术组、模型组、阳性对照组和实验组ColⅡ蛋白表达分别为0.11±0.02,0.01±0.00,0.07±0.01和0.08±0.01;这4组的MMP-9蛋白表达分别为0.46±0.16,3.79±1.11,0.97±0.38和0.95±0.31;这4组的MMP-13蛋白表达分别为0.43±0.16,5.97±0.84,0.89±0.67和0.98±0.56;这4组的TIMP-3蛋白表达分别为3.92±0.75,0.30±0.13,1.54±0.73和1.60±0.99。上述指标:模型组与假手术组比较,差异均有统计学意义(均P<0.01);阳性对照组、实验组与模型组比较,差异均有统计学意义(均P<0.01)。结论森登-4汤通过调节MMPS/TIMP通路达到修复软骨胶原纤维结构平衡,阻断KOA疾病的发展。

Objective To investigate the effect of Sundon-4 Decoction on theⅡtype collagen(ColⅡ),matrix metalloproteinases 9(MMP-9),tissue inhibitors of metalloproteinase 3(TIMP-3)in knee osteoarthritis(KOA)rabbits.Methods Rabbits KOA model was established by modified Hulth method.Rabbits were randomly divided into 4 groups according to weight:sham-operation group,model group,experimental group,positive control group,with 8 rabbits in each group.After the establishment of models,positive control group was given an intragastric Zhuanggu Joint Pill Suspension 0.51 g·kg^-1,while the experimental group was given Mongolian medicine Sendeng-4 Decoction 0.43 g·kg^-1.After four weeks intragastric administration,articular cartilage were took.The changes on ColⅡexpression(ratio of OD value)in collected joint cartilage tissue was detected by immunohistochemistry.The expression(ratio of gray value)in MMP-9,MMP-13,and TIMP-3 proteins was detected by Western blot.Results Expression of ColⅡprotein in sham-operation group,model group,positive control group,experimental group were 0.11±0.02,0.01±0.00,0.07±0.01 and 0.08±0.01;expression of MMP-9 protein in the four groups were 0.46±0.16,3.79±1.11,0.97±0.38 and 0.95±0.31;expression of MMP-13 protein in the four groups were 0.43±0.16,5.97±0.84,0.89±0.67 and 0.98±0.56;expression of TIMP-3 protein in the four groups were 3.92±0.75,0.30±0.13,1.54±0.73 and 1.60±0.99.Comparison between model group and sham-operation group,the difference of the factors were significant(all P<0.01);comparison between positive control group,experimental group and model group,the difference of the factors were significant(all P<0.01).Conclusion Sendeng-4 Decoction may repair cartilage collagen fiber structure balance by adjusting the MMPs/TIMP path,and finally blocks development of KOA disease.