基质金属蛋白酶组织抑制剂1对细胞因子诱导的胰岛β细胞凋亡的保护作用  

Protective Effect of Tissue Inhibitor of Metalloproteinase-1 on Apoptosis of Pancreatic β-Cells Induced by Cytokines

作　　者：周立娜[1] 刘倩琦[1] 寇敏[1] 朱敏[1] 潘晓勤[2] 

机构地区：[1]南京医科大学附属南京儿童医院内分泌科,南京210008 [2]南京医科大学第四临床医学院儿科医学研究所,南京210029

出　　处：《实用儿科临床杂志》2008年第20期1579-1581,共3页Journal of Applied Clinical Pediatrics

摘　　要：目的探讨基质金属蛋白酶抑制剂1(TIMP-1)对细胞因子IL-1β、TNF-α和干扰素γ(IFN-γ)诱导的胰岛β细胞凋亡的保护作用及其可能机制。方法实验分TIMP-1干预组、细胞因子刺激组和对照组。细胞因子刺激组应用细胞因子(IL-1β 10μg/L,TNF-α 50μg/L,IFN-γ 50μg/L)诱导大鼠胰岛瘤细胞株RINm5F细胞凋亡;TIMP-1干预组在细胞因子刺激前予以TIMP-1(50μg/L)预处理1 h;对照组采用无血清培养基同步培养。应用四甲基偶氮唑盐比色法和Caspase-3分光光度法检测各组细胞活力和Caspase-3活性,采用DNALadder进行凋亡鉴定。结果与对照组比较,细胞因子IL-1β、TNF-α和IFN-γ联合刺激组细胞活力明显降低,并呈时间依赖性(Pa<0.05,0.001);TIMP-1干预组较细胞因子刺激组细胞活力上升约14%,Caspase-3活性下降约38%,二组比较差异有显著性意义(Pa<0.001)。DNA Ladder检测,细胞因子刺激组可见特征性的梯状条带,TIMP-1干预组梯状条带不明显,出现与对照组相似的2条基因组大分子条带。结论IL-1β、TNF-α和IFN-γ联合刺激可诱导RINm5F细胞凋亡,TIMP-1对细胞因子诱导的RINm5F细胞的凋亡有一定的保护作用,此作用可能与其抑制Caspase-3活性有关。Objective To explore the effect and mechanism of tissue inhibitor of metalloproteinase - 1 （ TIMP - 1 ） on apoptosis of pancreatic β - cell induced by a combination of interleukin - 1β （ IL - 1β ）, tumor necrosis factor - α（ TNF -α） and interferon - γ （ IFN - γ）. Methods They were divided into 3 groups ：TIMP - 1 - treated group,cytokines - treated group and control group. The insulin - producing rat cell line RINm5F cells were cultured and stimulated by a combination of IL - 1β（ 10 ug/L） ,TNF -α（50 ug/L） ,and IFN - γ（50 ug/L） with or without the treatment of TIMP - 1 （50 ug/L） 1 hour before the addition of cytokines. The untreated control RINm5F cells were cultured in RMP11640 medium without serum. Cell viability,Caspase -3 activity and DNA fragmentation in RINmSF cells were measured after the cells exposed to cytokines plus or minus TIMP - 1 by methyl thiazolyl tetrazolium assay and spectrophotometrie method respectively. Apoptosis was assessed by DNA laddering experiment. Results Compared with control group, cell viability was decreased obviously in a time dependent manner in RINm5F cells after incubation with the cytokine mixture（P 〈 0.05,0. 001 ）. In the ceils after treatment with TIMP - 1, cell viability was remarkably increased （ by 14% , P 〈 0.05 ） and the activity of Caspase - 3 was significantly decreased （ by 38 % , P 〈 0. 001 ） compared with those in the cells treated by cytokines alone. Cytokines - treated cells showed classical DNA laddering,whereas cytokines plus TIMP - 1 treated cells showed complete inhibition of DNA laddering. Conclusions Exposure of RINm5 F cells to a combination of IL - 1 β, TNF - α, and IFN - γ can induce a significant apoptosis. TIMP - 1 can protect RINm5 F ceils against cytokine - induced apoptosis, which possibly involves in decreasing the activity of Caspase - 3.

关 键 词：胰岛Β细胞 凋亡 基质金属蛋白酶抑制剂1 细胞因子 大鼠 

分 类 号：R96[医药卫生—药理学]