阿仑膦酸钠对膝骨关节炎大鼠软骨细胞的保护作用

Effects of alendronate on chondrocytes in rats with knee osteoarthritis

目的探讨阿仑膦酸钠(ALN)对膝骨关节炎(KOA)大鼠软骨细胞的保护作用。方法用膝关节腔内注射弗氏完全佐剂(FCA)的方法建立KOA大鼠模型,将成功建模的40只KOA大鼠随机分为模型组(n=20)与实验组(n=20);另选择20只大鼠膝关节腔内注射生理盐水作为对照组。实验组大鼠给予ALN(20μg·kg^-1·2 d^-1)皮下注射,对照组与模型组大鼠分别给予等量生理盐水皮下注射,均连续给药12周。用TdT介导的dUTP缺口末端标记(TUNEL)检测软骨细胞凋亡指数(AI);用免疫组化法检测软骨组织基质金属蛋白酶-1(MMP-1)、基质金属蛋白酶组织抑制因子-1(TIMP-1)、Ⅱ型胶原蛋白(COL-Ⅱ)蛋白的表达情况。结果对照组、模型组与实验组大鼠软骨细胞AI分别为(9.34±2.89)%,(64.21±4.73)%和(23.65±3.82)%;MMP-1蛋白阳性细胞率分别为(18.64±2.31)%,(57.43±4.62)%和(26.38±3.74)%;TIMP-1蛋白阳性细胞率分别为(68.42±3.96)%,(23.51±2.62)%和(44.38±3.11)%;COL-Ⅱ蛋白阳性细胞率分别为(40.93±4.22)%,(25.36±3.02)%和(31.87±2.94)%。上述指标,模型组分别与对照组和实验组比较,差异均有统计学意义(均P<0.05)。结论ALN可显著抑制KOA大鼠炎症反应,降低软骨细胞凋亡,同时调节软骨组织MMP-1、TIMP-1、COL-Ⅱ蛋白表达,从而改善关节软骨病变。

Objective To investigate the effects of alendronate(ALN)on chondrocytes in rats with knee osteoarthritis(KOA).Methods The KOA rat models were established by injecting Freund’s complete adjuvant(FCA)into the knee joint cavity;40 KOA rats successfully established were randomly assigned to the model group(n=20)and the test group(n=20);another 20 rats were injected with saline in the knee joint cavity were assigned to the control group.Rats in the test group were given subcutaneous injection of ALN(20μg·kg^-1·2 d^-1),and rats in the control group and the model group were given subcutaneous injections of the same amount of normal saline for 12 weeks.TdT-mediated dUTP notch end labeling(TUNEL)was used to detect the chondrocyte apoptosis index(AI);The expression of matrix metalloproteinase-1(MMP-1),tissue inhibitor of matrix metalloproteinase-1(TIMP-1)and collagen type II(COL-Ⅱ)protein in cartilage tissues were detected by immunohistochemistry method.Results The chondrocyte AI in the control group,model group and test group were(9.34±2.89)%,(64.21±4.73)% and(23.65±3.82)%;The positive rates of MMP-1 protein cells in cartilage tissues were(18.64±2.31)%,(57.43±4.62)% and(26.38±3.74)%.The positive rates of TIMP-1 protein cells were(68.42±3.96)%,(23.51±2.62)% and(44.38±3.11)%;The positive rates of COL-Ⅱprotein cells were(40.93±4.22)%,(25.36±3.02)% and(31.87±2.94)%.There were statistically significant differences in the above indexes between the model group and the control group/test group(all P<0.05).Conclusion ALN can significantly inhibit the inflammatory response of KOA rats,reduce the apoptosis of chondrocytes,and regulate the expression of MMP-1,TIMP-1,COL-Ⅱ in cartilage tissues,thereby improving joint cartilage lesions.