高糖与同型半胱氨酸对大鼠皮肤成纤维细胞MMP-9表达及活性的影响  

Effects of high glucose and homocysteine on expression of MMP-9 in rat fibroblasts

作　　者：陈黎红[1] 谢晓英[1] 杨川[1] 严励[1] 朱平[1] 劳国娟[1] 郝少云[1] 

机构地区：[1]中山大学附属第二医院内分泌科,广东广州510120

出　　处：《中国病理生理杂志》2010年第9期1839-1843,共5页Chinese Journal of Pathophysiology

摘　　要：目的:观察不同浓度糖及同型半胱氨酸对大鼠皮肤成纤维细胞MMP-9表达量及活性的影响。方法:取1d龄正常SD大鼠背部皮肤分离成纤维细胞,于含10%胎牛血清的DMEM培养液中培养。2-4代生长状态良好的细胞进行24h血清剥夺(0.5%FBS)后,分为正常糖浓度组、高糖组、高糖+高同型半胱氨酸组分别培养6h,之后用RT-PCR、Western blotting和明胶酶谱方法分别检测MMP-9mRNA、蛋白表达量以及细胞外MMP-9活性。结果:高糖(22mmol/L)组MMP-9mRNA、蛋白表达量及其活性均增高,分别为正常糖浓度组的1.15倍、1.59倍和1.34倍,P<0.05;高糖+高同型半胱氨酸(100μmol/L)增高组更加明显,依次为正常糖浓度组的1.25倍、2.63倍和2.52倍,P<0.05。结论:大鼠皮肤成纤维细胞在体外高糖环境下可高表达MMP-9;高同型半胱氨酸在高糖环境下可使MMP-9表达及酶活性增高更为明显,表明高同型半胱氨酸血症也参与了对MMP-9的调节,可进一步增加高糖环境下成纤维细胞表达MMP-9。AIM： To observe the effect of different concentrations of glucose and homocysteine on the expression of MMP-9 in fibroblasts,and to investigate the relationship between homocysteine and diabetic foot. METHODS： Fibroblasts were obtained from the dermis of 1-day-old normal SD rat and cultured in DMEM containing 10% FBS. After deprivation of serum （ using 0. 5% FBS） for 24 h,the fibroblasts in 2-4 passages were cultured for 6 h under the conditions of normal glucose,high glucose and high glucose with high concentration of homocysteine,respectively. The expression of MMP-9 at mRNA and protein levels was assessed by RT-PCR and Western blotting,and the activity of MMP9 was determined by gelatin zymography analysis. RESULTS： The expression of MMP-9 at mRNA and protein levels and activity of MMP-9 in high glucose （ 22 mmol/L） group were 1. 15 folds,1. 59 folds and 1. 34 folds of those in normal glucose group （ P 0. 05） ,respectively. The effect of high glucose became more pronounced when co-treated with high concentration of homocysteine （ 100 μmol/L） ,which were 1. 25 folds,2. 63 folds and 2. 52 folds of those in normal glucose group （ P 0. 05） ,respectively. CONCLUSION： The increase in the expression of MMP-9 is concentration-dependen with glucose content in the culture. High concentration of homocysteine promotes this process,suggesting that hypercysteinemia might play an important role in the expression of MMP-9 in fibroblast.

关 键 词：高半胱氨酸 基质金属蛋白酶-9 成纤维细胞 

分 类 号：R322.99[医药卫生—人体解剖和组织胚胎学]