摘要
为探讨脂质过氧化损伤能否诱导血管内皮细胞表达细胞间粘附分子 1,用胰酶消化法分离人脐静脉内皮细胞进行原代培养 ,内皮细胞用不同浓度联胺作用相同时间或同一浓度联胺作用不同时间 ,使之发生脂质过氧化损伤 ,逆转录聚合酶链反应检测内皮细胞细胞间粘附分子 1mRNA表达 ,细胞酶链免疫吸附实验测定内皮细胞细胞间粘附分子 1蛋白表达。逆转录聚合酶链反应结果发现 ,不同浓度联胺 (1μmol L、5 μmol L和 10 μmol L)作用 8h ,内皮细胞细胞间粘附分子 1对 β actin吸光度比值依次为 0 .5 35、0 .90 8和 1.186 ,分别是对照组 (0 .185 )的 2 .89倍、4 .91倍和 6 .4 1倍 ;而同一浓度联胺 (5 μmol L)作用 4h和 2 4h ,吸光度比值依次为 0 .5 98和 1.2 92 ,分别是对照组 (0 .185 )的 3.2 3倍和 6 .98倍。细胞酶链免疫吸附实验结果发现 ,经 1μmol L、5 μmol L和 10 μmol L联胺作用 4h后 ,内皮细胞细胞间粘附分子 1蛋白表达的吸光度值依次为 0 .1387、0 .1775和 0 .2 32 6 ,分别是对照组 (0 .10 35 )的 1.34倍、1.71倍和 2 .2 5倍 (P <0 .0 1)。结果表明 ,联胺诱导的内皮细胞细胞间粘附分子 1的表达在一定作用时间和浓度范围内呈时间和剂量依赖性。提示联胺诱导的脂质过氧化损伤可能上调细胞间粘附分子
Aim To observe whether lipid peroxidation injury to endothelial cells (EC) induces intercellular adhesion molecule 1(ICAM 1) expression. Methods The lipid peroxidation injury to cultured human umbilical vein endothelial cells (hUVEC) was induced by exposure of the cells to diamide either at a same concentration for different incubation time or at different concentrations, but for a same incubation times. The ICAM 1 mRNA expression in hUVEC was determined by reverse transcriptase polymerase chain reaction (RT PCR), and was normalized with β actin mRNA. The ICAM 1 protein expression in hUVEC was determined by cell enzyme linked immunosorbent essy (ELISA). Results RT PCR showed that the ratios of the absorbance (A) values of ICAM 1 mRNA to that of the β actin mRNA expressed in hUVEC exposed to diamide at different concentrations (1 μmol/L, 5 μmol/L and 10 μmol/L) for 8 h was 0.535, 0.908 and 1.186, respectively. Whereas the ratio of the A values of ICAM 1 mRNA to that of the β actin mRNA expressed in hUVEC exposed to diamide at a concentration of 5 μmol/L for 0, 4 h and 24 h was 0.185, 0.598 and 1.292, respectively. Cell ELISA showed that the A values of ICAM 1 protein expressed in hUVEC exposed to diamide at different concentrations (1 μmol/L, 5 μmol/L and 10 μmol/L) for 4 h were 0.1387, 0.1775 and 0.2326, which were 1.34 fold, 1.71 fold and 2.25 fold as much as that of the control group (0.1035), respectively. There was a significant statistical difference between groups (F=38.89, P<0.01). Taken together, these results suggest that ICAM 1 expression in hUVEC induced by diamide was in a dose and time dependent manner. Conclusions Lipid peroxidation injury to EC induced by diamide may up regulate ICAM 1 mRNA and protein expression, which play an important role in the recruitment of monocytes into subendothelial space in atherogenesis.
出处
《中国动脉硬化杂志》
CAS
CSCD
2002年第2期109-111,共3页
Chinese Journal of Arteriosclerosis
基金
国家自然科学基金 (3 973 0 2 2 0 )
关键词
脂质过氧化
内皮
血管
细胞间粘附分子1
联胺
Lipid Peroxidation
Endothelium, Vascular
Intercellular Adhesion Molecule 1
Diamide
