脂多糖和同型半胱氨酸对人脐静脉内皮细胞单核细胞趋化蛋白1 mRNA表达的影响

目的探讨脂多糖和同型半胱氨酸是否诱导培养的人脐静脉内皮细胞表达单核细胞趋化蛋白1(MCP-1) mRNA及其机制.方法将生长至汇合的人脐静脉内皮细胞分为对照组、脂多糖组,脂多糖+SB203580组,同型半胱氨酸组,同型半胱氨酸+SB203580组,脂多糖+同型半胱氨酸组.采用斑点杂交和RT-PCR检测其MCP-1 mRNA的表达.用免疫细胞化学法检测对照组、脂多糖组和同型半胱氨酸组P38蛋白激酶蛋白的表达.结果培养的人脐静脉内皮细胞能表达较低水平的MCP-1 mRNA.斑点杂交和RT-PCR均显示各实验组的MCP-1 mRNA明显高于对照组.免疫细胞化学显示,P38蛋白激酶蛋白在对照组的细胞核阳性表达率为9.6%,当人脐静脉内皮细胞暴露于脂多糖或同型半胱氨酸后,细胞核阳性表达率升至46.7%或57.7%.结论脂多糖和同型半胱氨酸能诱导人脐静脉内皮细胞表达单核细胞趋化蛋白1 mRNA,P38蛋白激酶可能参与了该过程.

Lipopolysaccharide and homocysteine induce expression of monocyte chemoattractant protein-1 mRNA in human umbilical vein endothelial cells

Aim:To investigate whether lipopolysaccharide (LPS) and homocysteine (HCY) could induce cultured human umbilical vein endothelial cells (HUVECs) to express monocyte chemoattractant protein 1 (MCP 1) mRNA and the mechanism.Methods: The confluent HUVECs were allocated into 6 groups: control group, LPS group, LPS+SB203580 group, HCY group, HCY+SB203580 group, and LPS+HCY group. The expression of MCP 1 mRNA in HUVECs was assayed by dot hybridization and RT PCR. The expressions of P38 protein kinase protein in HUVECs of the control group, LPS group and HCY group were measured by immunocytochemistry.Results:The cultured HUVECs expressed MCP 1 mRNA at a low level. Both dot hybridization and RT PCR showed that the expressions of MCP 1 mRNA of all experimental groups were higher than that of control group. The immunocytochemistry showed that the percentage of nuclear positive cells of P38 in the control group was 9.6%, after exposure of HUVECs to HCY and LPS, it increased to 46.7% and 57.7%, respectively.Conclusion:LPS and HCY could induce the expression of MCP 1 mRNA in HUVECs, and P38 protein kinase may play an important role in the process.