高糖对基质金属蛋白酶2及其抑制剂在人腹膜间皮细胞中表达的影响

目的 研究高糖对人腹膜间皮细胞(HPMC)基质金属蛋白酶2(MMP2)及其组织抑制剂(TIMP)1、TIMP2基因及蛋白表达的影响,以及对人腹膜间皮下细胞外基质(ECM)降解的可能作用。方法 利用腹膜透析(腹透)患者腹透流出液标本原代培养HPMC并进行传代,采用半定量RT-PCR方法研究高糖高渗对HPMC的MMP2、TIMP1、TIMP2基因表达的影响。利用免疫组织化学(组化)及Zymography方法检测HPMC的MMP2、TIMP1、TIMP2蛋白表达。结果 HPMC存在MMP2、TIMP1、TIMP2基因及蛋白表达,4.25%葡萄糖显著上调HPMC的TIMP1基因表达(P<0.01),高渗对HPMC的TIMP1基因表达无明显影响(P>0.05),高糖高渗对HPMC的MMP2、TIMP2基因表达无明显影响(P>0.05)。结论 HPMC能够通过MMP/TIMP系统影响到腹膜ECM的降解,高糖可能通过诱导HPMC的TIMP1与MMP间表达失衡,在促进腹膜纤维化的进展中发挥一定作用。

Effect of high glucose on the expression of matrix metalloproteinase 2 and its inhibitors in human peritoneal mesothelial cells

Objective To study the gene and protein expression of matrix metalloproteinase 2 (MMP2) and its inhibitors TIMP1 and TIMP2 in human peritoneal mesothelial cells (HPMC), and the possible role of high glucose in submesothelial extracellular matrix (ECM) degradation during peritoneal dialysis (PD) . Methods Primary HPMC was isolated from spent peritoneal dialysis effluent collected from PD patients. After HPMC confluence, the cells were detached by trypsinization and passaged into 25 cm2 tissue-culture flasks. The effect of high glucose and hyperosmolarity on the gene expression of MMP2, TIMP1 and TIMP2 in HPMC was studied by semi-quantitative RT-PCR. Immunohistochemistry and zymography were used to measure the protein expression of MMP/TIMP in HPMC. Results HPMC expressed MMP2, TIMP1, TIMP2 at both gene and protein levels. 4. 25% glucose significantly up-regulated TIMP1 gene expression in HPMC( P < 0. 01). Hyperosmolarity did not affect TIMP1 gene expression. MMP2 and TIMP2 gene expression was not affected by high glucose or hyperosmolarity. Conclusions HPMC can influence submesothelial ECM remodeling through the expression of MMP/TIMP. High glucose may induce the imbalance between TIMP1 and MMP expression in HPMC, thus in part accelerate the progression of peritoneal fibrosis during dialysis.