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目的 探讨同型半胱氨酸对人单核细胞株THP 1表达趋化因子受体CCR1mRNA的影响。方法 在THP 1单核细胞培养基中加入不同浓度的同型半胱氨酸 ,孵育 8h ;或加入终浓度 0 1mmol/L的同型半胱氨酸 ,分别孵育 4、8、 16h ,再分别用逆转录多聚酶链反应和原位杂交检测各组THP 1单核细胞CCR1mRNA。结果 对照组THP 1单核细胞表达较低水平的CCR1mRNA ,加同型半胱氨酸后各实验组CCR1mRNA表达随浓度和时间的增加而增强 ,且PCR产物经测序证实。原位杂交显示 ,THP 1单核细胞表达CCR1mRNA为胞质内紫蓝色颗粒。图像分析显示 ,经同型半胱氨酸处理后 ,各组细胞胞质内CCR1mRNA表达明显增加 ,并呈剂量和时间依赖性。方差分析表明 ,组间差异极为显著(P <0 0 1)。结论 同型半胱氨酸能诱导THP 1单核细胞表达CCR1mRNA ,且与同型半胱氨酸的浓度和时间呈正相关 ,并可能通过促进招募单核细胞进入内皮下间隙 ,在动脉粥样硬化发病机制中起重要作用
Objective To investigate the effect of homocysteine on chemokine receptor CCR1 mRNA expression in THP1 human monocytic cell line. Methods Different concentrations of homocysteine were added into THP 1 monocyte culture medium and incubated for 8 hours. After the cells were incubated with 0 1 mmol / L homocysteine for 4, 8 and 16 hours respectively, Polymerase chain reaction and in situ hybridization were used to detect CCR1 mRNA in THP1 monocytes. Results The expression of CCR1 mRNA in THP-1 monocytes was lower in control group than that in control group. The expression of CCR1 mRNA in each experimental group increased significantly with the increase of concentration and time, and the PCR products were confirmed by sequencing. In situ hybridization showed that THP1 monocytes expressed CCR1 mRNA as intracytoplasmic purple-blue granules. Image analysis showed that after homocysteine treatment, the expression of CCR1mRNA in the cytoplasm of each group increased obviously, in a dose-and time-dependent manner. Analysis of variance showed that there was a significant difference between groups (P <0.01). CONCLUSION: Homocysteine can induce the expression of CCR1 mRNA in THP 1 monocytes, which is positively correlated with the concentration and time of homocysteine and may promote the expression of CCR1 in the pathogenesis of atherosclerosis by promoting the recruitment of monocytes into the subepithelial space Play an important role