大蒜素对SD大鼠膝骨关节炎关节软骨中MMP-13、TIMP1和Collagen Ⅱ表达的影响

Effect of allicin on the expression of MMP-13, TIMP1, and collagen Ⅱ in SD rat knee cartilage with osteoarthritis

目的探讨大蒜素对SD大鼠膝骨关节炎关节软骨中基质金属蛋白酶-13(MMP-13)、基质金属蛋白酶组织抑制因子1(TIMP1)和胶原蛋白酶Ⅱ(CollagenⅡ)的影响,明确大蒜素对SD大鼠膝骨关节炎关节软骨的保护作用及机制。方法 90只SD大鼠随机分为5组:空白对照组、手术对照组、手术组+5 mg大蒜素、手术组+10 mg大蒜素、手术组+20 mg大蒜素。手术对照组和手术组均行改良Hulth法复制OA模型,于术后第2日开始分组腹腔注射给药:空白对照组和手术对照组给予等量生理盐水腹腔注射,另外3组分别给予大蒜素5、10、20 mg/(kg·d)腹腔注射。用药干预12周后处死,取软骨组织,运用Western blot和免疫组化实验检测MMP-13、TIMP1和CollagenⅡ表达。结果Western blot检测发现大蒜素10和20 mg/(kg·d)给药12周后能够下调MMP-13的表达,上调TIMP1的表达,同时2组中CollagenⅡ的表达较手术对照组及手术+5 mg大蒜素组多;免疫组化染色结果进一步明确MMP-13、TIMP1和CollagenⅡ在软骨组织中的表达,变化趋势与Western blot一致。结论大蒜素通过下调关节软骨中MMP-13的表达,上调关节软骨中TIMP1及CollagenⅡ的表达,减少细胞外基质的降解,对SD大鼠骨性关节炎中关节软骨具有积极保护作用。

Objective To explore the effect of allicin on the expression of matrix metalloproteinase-13（MMP-13）, tissue inhibitor of metalloproteases 1（TIMP1） and Collagen Ⅱ in SD rat knee cartilages with osteoarthritis. Methods Totally 90 SD rats were randomly diveded into 5 groups： a blank control group, a surgery control group, a surgery ＋ 5 mg allicin group, a surgery ＋ 10 mg allicin group, and a surgery ＋ 20 mg allicin group. After the surgery, the surgery control group and the surgery group had a modified Hulth modelling surgery on their knee joints. The blank control group and the surgery control group were treated with normal saline at the same dose. The surgery group was treated with allicin（5 mg, 10 mg, 20 mg）. The proteins expression of MMP-13, TIMP1 and Collagen Ⅱ were determined by Western blot. The expression level of MMP-13, TIMP1 and Collagen Ⅱ in the cartilages were evaluated by immunohistochemistry. Results Western blot demonstrated allicin markedly downregulated the expression of MMP-13, and increased the expression of Collagen Ⅱ and TIMP1. Immunohistochemistry assay found that allicin significantly inhibited the expression of MMP-13, and increased the expression of Collagen Ⅱ and TIMP1. Conclusion Allicin has a protective effect on SD rat knee cartilages with osteoarthritis increasing the expression of TIMP1 and Collagen Ⅱ, as well as decreasing MMP-13 expression.