高糖对大鼠皮肤成纤维细胞生长及胶原蛋白合成的影响  

Effects of high concentration of glucose on growth and collagen synthesis of rat skin fibroblast cells

作　　者：张再超[1] 叶希韵[1] 刘霞[1] 翁宇静[1] 

机构地区：[1]华东师范大学生命科学学院生命医学系,上海200062

出　　处：《上海交通大学学报（医学版）》2009年第3期323-326,共4页Journal of Shanghai Jiao tong University:Medical Science

摘　　要：目的研究不同浓度的葡萄糖对体外培养的大鼠皮肤成纤维细胞生长及胶原蛋白合成的影响。方法体外培养的大鼠皮肤成纤维细胞中加入含不同浓度葡萄糖培养液(30、35、40mmol/L)培养48h(高糖1～3组),用MTT法测定细胞生长情况;试剂盒法测定细胞培养液中羟脯氨酸质量-体积浓度;RT-PCR技术检测Ⅰ、Ⅲ型前胶原蛋白以及基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶抑制酶(TIMP-2)mRNA的表达。以加入含25mmol/L葡萄糖培养液的细胞作为正常对照组。结果随着培养液中葡萄糖浓度的升高,皮肤成纤维细胞的生长明显受抑;培养液中羟脯氨酸的质量-体积浓度明显降低(P<0.01)。RT-PCR结果显示,高糖组细胞内Ⅰ、Ⅲ型前胶原蛋白及TIMP-2 mRNA表达明显下调,MMP-2 mRNA表达明显上调,与正常对照组比较差异显著(均P<0.05)。结论高浓度葡萄糖对体外培养的大鼠皮肤成纤维细胞的生长有一定的抑制作用,并可能通过抑制Ⅰ、Ⅲ型前胶原蛋白、TIMP-2和上调MMP-2的mRNA表达导致皮肤成纤维细胞胶原蛋白的合成减少。Objective To investigate the effects of different concentrations of glucose on growth and collagen synthesis of rat skin fibroblast cells cultured in vitro. Methods Rat skin fibroblast cells cultured in vitro were treated with glucose of different concentrations （30,35 and 40mmol/L） for 48 h （high glucose group 1,2 and 3）. Cell proliferation was detected by MTT method,the hydroxyproline contents in culture media were determined by the commercial kit,and the mRNA expression of typeⅠand Ⅲ procollagen,matrix metalloproteinase-2（MMP-2）and tissue inhibitor of metalloproteinase-2（TIMP-2）were detected by RT-PCR. Cells cultured with 25mmol/L glucose were served as controls. Results With the increase of glucose concentration in culture media,the growth of skin fibroblast cells was significantly inhibited,and the hydroxyproline contents were significantly decreased （P〈0.01）. Compared with the controls,the mRNA expression of type Ⅰand Ⅲ procollagen and TIMP-2 were significantly decreased,while the mRNA expression of MMP-2 was significantly increased in high glucose group （P〈0.05）. Conclusion High concentration of glucose can significantly inhibit the growth of rat skin fibroblasts cultured in vitro,and both down-regulation of typeⅠprocollagen,type Ⅲ procollagen and TIMP-2 mRNA expression and up-regulation of MMP-2 mRNA expression may contribute to the decrease of collagen synthesis.

关 键 词：皮肤成纤维细胞 高糖 RT-PCR Ⅰ型胶原蛋白 Ⅲ型胶原蛋白 基质金属蛋白酶 基质金属蛋白酶抑制酶 

分 类 号：R361.2[医药卫生—病理学]