摘要
目的观察阿仑膦酸钠对长期应用皮质激素大鼠骨组织中基质金属蛋白酶-2(MMP-2)、MMP-9及其特异性抑制因子基质金属蛋白酶组织抑制剂-1(TIMP-1)和TIMP-2mRNA表达的影响,探讨阿仑膦酸钠预防激素性股骨头坏死的效果及其作用机制。方法健康SD大鼠30只,雌雄各半,随机分为激素组、阿仑膦酸钠组和空白对照组,每组10只。激素组和阿仑膦酸钠组肌内注射醋酸泼尼松龙12.5mg/kg,每周2次,阿仑膦酸钠组同时给予阿仑膦酸钠5mg/kg灌胃,每周1次;对照组只给予相同体积生理盐水肌注。4周后取左侧股骨头骨组织石蜡包埋,HE染色,鉴定骨质疏松和股骨头坏死情况;取右侧股骨头骨组织提取总RNA,采用逆转录聚合酶链反应(RT-PCR)技术检测MMP-2、MMP-9、TIMP-1和TIMP-2的mRNA表达水平。结果 MMP-2在激素组、阿仑膦酸钠组、对照组中的表达分别为0.15±0.04、0.13±0.07、0.09±0.03,MMP-9在三组中的表达分别为0.13±0.03,0.09±0.02和0.08±0.02;TIMP-1在三组中的表达分别为0.07±0.02,0.13±0.4和0.18±0.04;TIMP-2在三组中的表达分别为0.45±0.15,0.63±0.10和0.69±0.19。激素组与对照组比较,MMP-2、MMP-9mRNA表达增高,TIMP-1、TIMP-2mRNA表达降低,差异有统计学意义(P<0.05)。阿仑膦酸钠组与对照组比较,仅MMP-2显著性差异。阿仑膦酸钠组与激素组比较,MMP-2、MMP-9mRNA表达降低,TIMP-1、TIMP-2mRNA表达增加,差异有统计学意义(P<0.05)。结论醋酸泼尼松龙上调大鼠骨组织中MMP-2、MMP-9mRNA的表达,下调TIMP-1、TIMP-2水平,使MMPs/TIMPs比值升高。阿仑膦酸钠可以拮抗皮质激素对MMPs/TIMPs系统的调控。
Objective To investigate the effects of alendronate on matrix metalloproteinase-2（MMP-2）,matrix metalloproteinase-9（MMP-9） and tissue inhibitor of matrix metalloproteinases-1（TIMP-1）,tissue inhibitor of matrix metalloproteinases-2（TIMP-2） mRNAs expression in bone tissues of femoral head of rats using glucocorticoid for long time,and to discuss the mechanism of alendronate in preventing glucocorticoid femoral head necrosis.Methods Thirty adult SD rats were randomly divided into glucocorticoid group,alendronate group and control group.The glucocorticoid group and alendronate group were treated by prednisolone intramuscular injection,12.5 mg/kg,two times per week,and the alendronate group was administered with alendronate 5 mg/kg by intragastric at the same time,one time per week.The control group was treated only by the same volume sodium chloride intramuscular injection.After 4 weeks,the osteonecrosis of left femoral head was detected by HE stain,the total RNA of right femoral head bone tissue was extracted and the expression levels of MMP-2,MMP-9,TIMP-1 and TIMP-2 mRNAs were examined by RT-PCR.Results The mortality of rats were 10%（1/10）,10%（1/10）,0（0/10） in glucocorticoid,alendronate and control group.The necrosis rate of femoral bone head were 66.67%（6/9）,22.22%（2/9） and 0（0/10）.The expression of MMP-2 in hormone group,alendronate group and control group were 0.15 ± 0.04,0.13 ± 0.07 and 0.09 ± 0.03.The expression of MMP-9 in the 3 groups were 0.13 ± 0.03,0.09 ± 0.02 and 0.08 ± 0.02.The expression of TIMP-1 in the 3 groups were 0.07 ± 0.02,0.13 ± 0.4 and 0.18 ± 0.04.The expression of TIMP-2 in the 3 groups were 0.45 ± 0.15,0.63 ± 0.10 and 0.69 ± 0.19.Compared with control group,MMP-2,MMP-9 mRNA expression increased significantly,and TIMP-1,TIMP-2 mRNA expression decreased significantly in glucocorticoid group（P〈0.05）.MMP-2 expression was significantly different between alendronate group and control group.Compared with the glucocorticoid group,MMP-2,MMP-9 mRNA expression decreased,and TIMP-1,TIMP-2 mRNA expression increased significantly in alendronate group（P〈0.05）.Conlusion Glucocorticoid significantly up-regulates MMP-2,MMP-9 mRNA and down-regulates the expression of TIMP-1,TIMP-2 mRNA.But this change can be prevented by alendronate.
出处
《中国骨与关节外科》
2011年第6期484-489,共6页
Chinese Journal of Bone and Joint Surgery
基金
国家自然科学基金资助项目(No.81160228):OPG/RANKL/RANK基因多态性与蒙汉两族人群激素性骨坏死和酒精性骨坏死的关联性及淫羊藿提取物个体化防治研究
