摘要
为探讨天然及氧化型极低密度脂蛋白能否诱导培养的人脐静脉内皮细胞表达巨噬细胞炎性蛋白 1α ,使内皮细胞分别暴露于上述脂蛋白后 ,用原位杂交、逆转录聚合酶链反应及免疫细胞化学法分别检测各组细胞的巨噬细胞炎性蛋白 1αmRNA和蛋白的表达。原位杂交发现 ,培养的内皮细胞能表达巨噬细胞炎性蛋白 1αmRNA ,两脂蛋白组内皮细胞的积分吸光度值明显高于对照组 (P <0 .0 1)。逆转录聚合酶链反应发现 ,两脂蛋白组内皮细胞的巨噬细胞炎性蛋白 1αmRNA的积分吸光度值分别为对照组的 15 .4倍和 14 .2倍。免疫细胞化学发现 ,两脂蛋白组内皮细胞胞浆的巨噬细胞炎性蛋白 1α蛋白表达 (棕色颗粒 )的积分吸光度值显著高于对照组 ,方差分析表明 ,组间差异有显著性意义 (P <0 .0 5 )。结果提示 ,天然及氧化型极低密度脂蛋白均可诱导培养的人脐静脉内皮细胞表达高水平的巨噬细胞炎性蛋白 1αmRNA和蛋白 。
Aim To understand whether native very low density lipoprotein (n VLDL) and oxidized very low density lipoprotein (ox VLDL) induce the cultured human umbilical endothelial cells (EC) to express macrophage inflammatory protein 1α (MIP 1α). Methods The EC were cultured in serum free medium containing 80 mg/L of n VLDL and ox VLDL respectively, while the control group without lipoproteins. After 24 h incubation, the EC grown on the cover slips were fixed and in situ hybridized with the Digoxigenin labeled MIP 1α cDNA probe. The MIP 1α mRNA expressed by the cells of every group was determined by reverse transcription polymerase chain reaction (RT PCR) as well. The MIP 1α protein expressed by the cells was detected by immunocytochemistry. Results Densitometry scan showed that the cultured EC could express MIP 1α mRNA, and the integral absorbance (A) values of the cells exposed to lipoproteins in situ hybridized with the probe metioned above were significantly higher than that of the control group. Analysis of variance demonstrated that there was significant difference among groups (P<0.01). RT PCR also showed that the A values of the MIP 1α mRNA expressed by the cells in n VLDL and ox VLDL group were 15.4 and 14.2 fold as much as that of the control group. Immunocytochemistry showed that the A values of the MIP 1α protein expression in the cells (brown granular substance) in n VLDL and ox VLDL group were significant higher than that of the control group. Analysis of variance showed a significant difference among groups (P<0.05). Conclusion n VLDL and ox VLDL were able to induce MIP 1α mRNA and protein expression in human umbilical vein endothelial cells at a high level, and may play an important role in atherogenesis through enhancing recruitment of monocytes to the intuma.
出处
《中国动脉硬化杂志》
CAS
CSCD
2002年第1期13-15,共3页
Chinese Journal of Arteriosclerosis
基金
国家自然科学基金 (39730 2 2 0 )资助
