oxLDL通过CXCL16、ERK1/2信号通路在川崎病冠状动脉损伤中的研究

The role of oxLDL in coronary artery lesion in Kawasaki disease through CXCL16 and ERK1/2 signaling pathways

目的探究氧化低密度脂蛋白(oxLDL)通过CXCL16/ERK1/2信号通路与川崎病(KD)冠状动脉损伤(CAL)的关系。方法构建KD的细胞模型,将实验分为3组:正常对照组(Con组)、KD冠状动脉正常组(NCAL组,加入10%NCAL患者血清)、KD冠状动脉损伤组(CAL组,加入10%CAL患者血清)。细胞培养24 h后,利用Elisa法检测细胞上清液中oxLDL的表达水平;另据公共数据库筛选oxLDL对KD病情的潜在相关靶点,用RT-qPCR检测CXCL16的表达水平;采用Western Blot技术检测细胞蛋白CXCL16、p-ERK的表达水平;通过CCK8试验和Edu法评估内皮细胞的活力和增殖活性,利用内皮细胞划痕试验检测细胞迁移能力。运用ERK1/2抑制剂(SCH772984)干预ERK1/2表达,建立了四个不同处理组别:NCAL组、NCAL+SCH组、CAL组、CAL+SCH组,并通过Western Blot技术检测CXCL16、p-ERK的表达水平变化。结果与Con组相比,NCAL组中oxLDL和CXCL16的表达水平较高,而在CAL组中这一趋势更为显著(P<0.05)。相较于Con组,NCAL组中的CXCL16、p-ERK的表达呈上升趋势,而在CAL组中这种趋势进一步增强(P<0.05)。CAL组内皮细胞死亡明显,CCK-8检测显示CAL患者血清刺激后细胞活性较低(P<0.05),Edu检测结果显示CAL组的Edu阳性细胞显著减少(P<0.05),细胞划痕愈合速度减缓(P<0.05)。结果显示抑制ERK1/2组在抑制ERK1/2表达后CXCL16的表达明显下降(P<0.05)。结论oxLDL可能透过CXCL16/ERK1/2信号通路加重内皮细胞功能受损,从而引发KD患者CAL的发生。

Objective To investigate the relationship between oxidized low density lipoprotein(oxLDL)and coronary artery lesion(CAL)in Kawasaki disease(KD)through CXCL16/ERK1/2 signaling pathway.Methods Cell models of KD were established and divided into three groups:normal control group(Con group),normal coronary artery group(NCAL group,10%serum from NCAL patients),and coronary artery lesion group(CAL group,10%serum from CAL patients).After 24 hours of cell culture,the expression level of oxLDL in the cell supernatant was detected by Elisa.In addition,the potential targets of oxLDL in KD were screened according to the public database,and the expression level of CXCL16 was detected by RT-qPCR.The expression level of cell protein CXCL16 and p-ERK were detected by Western Blot.CCK8 assay and Edu assay were used to evaluate the viability and proliferation of endothelial cells,and scratch assay was used to detect the migration ability of endothelial cells.The expression of ERK1/2 was interfered by ERK1/2 inhibitor(SCH772984),and four different treatment groups were established:NCAL group,NCAL+SCH group,CAL group and CAL+SCH group.The expression levels of CXCL16 and p-ERK were detected by Western Blot.Results Compared with Con group,the oxLDL and the expression of CXCL16 levels were higher in the NCAL group,but the trend was more significant in the CAL group(P<0.05).Compared with the Con group,the expression of CXCL16 and p-ERK in the NCAL group showed an upward trend,and this trend was further enhanced in the CAL group(P<0.05).CCK-8 test showed that the cell activity was lower after stimulation with the serum of CAL patients(P<0.05).Edu test results showed that the number of Edu positive cells in the CAL group was significantly reduced(P<0.05),and the cell scratch healing speed was slowed down(P<0.05).The results showed that the expression of CXCL16 in the inhibition of ERK1/2 group was significantly decreased after the inhibition of ERK1/2 expression(P<0.05).Conclusion oxLDL may aggravate the dysfunction of endothelial cells through CXCL16/ERK1/2 signaling pathway,thereby triggering the occurrence of CAL in KD patients.