摘要
为探讨活化的淋巴细胞直接接触对血管平滑肌细胞基质金属蛋白酶表达的影响 ,用乙酸肉豆蔻佛波醇活化并经 1%多聚甲醛固定的人外周血淋巴细胞 ,与培养的人类胚胎血管平滑肌细胞共同孵育 ,用聚丙烯酰胺凝胶电泳法测定细胞培养上清液中基质金属蛋白酶的活性。结果发现 ,基础状态下的体外培养人血管平滑肌细胞合成基质金属蛋白酶 2的酶原形式 ,有少量基质金属蛋白酶 2的活性形式存在。经佛波醇活化淋巴细胞组上清液中可检测到基质金属蛋白酶 1、基质金属蛋白酶 3和基质金属蛋白酶 9活性 ,并且基质金属蛋白酶 2活性形式较对照组增加 198%± 2 7% (P <0 .0 1) ,未经佛波醇活化的淋巴细胞不影响血管平滑肌细胞基质金属蛋白酶的合成和分泌。以上结果提示 ,活化固定的淋巴细胞可以通过细胞直接接触诱导血管平滑肌细胞基质金属蛋白酶表达增加及活化 。
Aim To Investigate whether activate lymphocyte contact induces the expression of matrix metalloproteinases in vascular smooth muscle cells. Methods The healthy human peripheral blood lymphocytes, which were activated by incubating with PMA and then fixed in 1% paraformaldehyde, were incubated with cultured human fetal aortic vascular smooth muscle cells (hVSMC). The gelatinase (MMP-2 and MMP-9) and stromelysin (MMP-3) activity in conditioned culture media were demonstrated by gelatin SDS-PAGE and casein SDS-PAGE. Results Unstimulated cultured hVSMC constitutively express 72 kDa zymogen of MMP-2 (pro-MMP-2) with a little of 66 kDa activated MMP-2, and no MMP-1, MMP-3, MMP-9 were detected by SDS-PAGE. After incubating with PMA-stimulated lymphocytes for 24 hours, zymography showed the MMP-1, MMP-3 and MMP-9 activity in the conditioned culture media, and activated MMP-2 increased 198%±27% compared with the control group (P<0.01). The unstimulated lymphocytes had no effect on MMPs synthesis of human fetal aortic vascular smooth muscle cells and PMA-activated lymphocyte had no gelatinolytic and caseinolytic activity on its own. Conclusions Activated lymphocytes may play an important role in atherosclerotic plaque repture through inducing vascular smooth cells to produce increased MMP and activate MMP-2.
出处
《中国动脉硬化杂志》
CAS
CSCD
2002年第4期300-303,共4页
Chinese Journal of Arteriosclerosis
基金
天津市科委重点课题资助