Inhibition of apoptosis in rat mesangial cells by tissue inhibitor of metalloproteinase-1
Inhibition of apoptosis in rat mesangial cells by tissue inhibitor of metalloproteinase-1
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DOI：
10.1046/j.1523-1755.2002.00403.x
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发表时间：
2002-07-01
影响因子：
19.6
通讯作者：
Yu, ZH
中科院分区：
文献类型：
--
作者：
Lin, HL;Chen, XM;Yu, ZH
背景金属蛋白酶组织抑制剂-1（TIMP-1）是细胞外基质降解的重要抑制剂。近年来，有研究表明TIMP-1还能抑制B型淋巴细胞的凋亡。本研究旨在探讨TIMP-1对肾小球系膜细胞增殖的影响。克隆了TIMP-1的全长cDNA，构建了TIMP-1 S和TIMP-1AS两个重组载体，分别编码正义TIMP-1和反义TIMP-1。转染大鼠肾小球系膜细胞（RMC），用北方和Western blotting检测其表达。通过血清剥夺诱导细胞凋亡，并通过TUNEL法和DNA梯状电泳监测DNA片段化。同时检测内源性TIMP-1、基质金属蛋白酶-2（MMP-2）、MMP-9及凋亡相关基因Bcl-2、Bax的表达。TIMP-1AS转染诱导TIMP-1表达的抑制，伴随着细胞凋亡的早期发生，TIMP-1。S转染诱导TIMP-1的过度表达，伴随着一个更晚的发病凋亡。TIMP-1的中和抗体恢复TIMP-1 S转染的RMC对血清剥夺的敏感性，但合成的基质金属蛋白酶抑制剂BB-94不影响TIMP-1 S转染的RMC对血清剥夺的敏感性。TIMP-1过表达可抑制Bax的表达，但对Bcl-2的表达无影响。TIMP-1抑制血清剥夺诱导的RMC凋亡，Bax可能参与了TIMP-1的作用。
Background. Tissue inhibitor of metalloproteinase-1 (TIMP-1) is an important inhibitor of extracellular matrix degradation. Recently, it was reported that TIMP-1 also could inhibit apoptosis in B type lymphocyte. This study was designed to examine the effects of TIMP-1 on mesangial cell apoptosis.Methods. The full-length cDNA of TIMP-1 was cloned and used to construct two recombinant vectors, TIMP-1S and TIMP-1AS, encoding sense TIMP-1 and antisense TIMP-1, respectively. The vectors were transfected into rat mesangial cells (RMC) and their expressions detected by Northern and Western blotting. Apoptosis was induced by serum deprivation, and was monitored for DNA fragmentation by TUNEL assay and DNA laddering. In addition, the expression of endogenous TIMP-1, matrix metalloprotein-2 (MMP-2), and MMP-9, as well as apoptosis-related genes Bcl-2 and Bax were investigated.Results. TIMP-1AS transfection induced a suppression of TIMP-1 expression accompanied by an earlier onset of apoptosis, and TIMP-1. S transfection induced TIMP-1 over-expression accompanied by a much later onset of apoptosis. A neutralizing antibody of TIMP-1 restored the sensitivity of TIMP-1S-transfected RMC to serum deprivation, but a synthetic matrix metalloproteinase inhibitor BB-94 did not influence the sensitivity of TIMP-1S-transfected RMC to serum deprivation. Finally, TIMP-1 over-expression inhibited the expression of Bax but with no effect on the expression of Bcl-2.Conclusion. TIMP-1 inhibits the serum deprivation-induced apoptosis in RMC, in which Bax might be involved.