LY294002对类脂A诱导的人冠状动脉内皮细胞缝隙连接蛋白43表达的影响

Effects of LY294002 on expression of connexin 43 in human coronary artery endothelial cells induced by lipoid A

目的探讨LY294002对类脂A(KLA)诱导的人冠状动脉内皮细胞(HCAEC)缝隙连接蛋白43(Cx43)表达的影响。方法用CCK8法检测0、0.01、0.05、0.10、0.20 mg/L KLA和0、2.5、5.0、10.0、20.0μmol/L磷酸肌醇3激酶(PI3K)特异性抑制剂——LY294002对HCAEC活力的影响。将HCAEC采用简单随机分组法分为空白对照组(Con组)、Toll样受体4(TLR4)激动组(KLA组)、PI3K抑制组(LY组)和KLA+LY组,采用Western Blot法检测各组HCAEC TLR4、PI3K、Cx43蛋白表达水平。结果0.01、0.05 mg/L KLA HCAEC存活率与0 mg/L比较,2.5μmol/L LY294002 HCAEC存活率与0μmol/L比较,差异均无统计学意义(P>0.05);0.10、0.20 mg/L KLA HCAEC存活率均明显低于0 mg/L,5.0、10.0、20.0μmol/L LY294002 HCAEC存活率均明显低于0μmol/L,差异均有统计学意义(P<0.05)。KLA组HCAEC TLR4、PI3K、Cx43蛋白表达水平明显高于Con组,LY组HCAEC PI3K蛋白表达水平明显低于Con组,KLA+LY组HCAEC TLR4、PI3K、Cx43蛋白表达水平均低于KLA组,差异均有统计学意义(P<0.05);LY组HCAEC TLR4、Cx43蛋白表达水平与Con组比较,差异均无统计学意义(P>0.05)。结论LY294002对正常培养状态下HCAEC TLR4、Cx43蛋白表达无显著影响。LY294002可抑制由KLA诱导的HCAEC Cx43高表达。

Objective To investigate the effect of LY294002 on the expression of connexin 43(Cx43)in human coronary endothelial cells(HCAEC)induced by lipoid A(KLA).Methods The effects of 0,0.01,0.05,0.10,0.20 mg/L of KLA and 0,2.5,5.0,10.0,20.0μmol/L of PI3K specific inhibitor LY294002 on the HCAEC activity were detected by CCK8 assay.HCAEC were randomly divided into the blank control group(Con group),TLR4 agonist group(KLA group),PI3K inhibition group(LY group)and KLA+LY group.The protein expression levels of TLR4,PI3K and Cx43 in HCAEC of each group were detected by Western Blot.Results The survival rate of HCAEC had no statistical difference between 0.01,0.05 mg/L KLA and 0 mg/L KLA,and between 2.5μmol/L LY294002 and 0μmol/L LY294002(P>0.05),while the survival rate of HCAEC in 0.10,0.20 mg/L KLA was significantly lower than that of 0 mg/L KLA(P<0.05).The survival rates of HCAEC in 5.0,10.0,20.0μmol/L LY294002 were significantly lower than those in 0μmol/L,and the differences were statistically significant(P<0.05).The protein expression levels of TLR4,PI3K and Cx43 in HCAEC of the TLR4 group were significantly higher than those in the Con group,the P13K protein expression level of HCAEC in the LY group were significantly lower than those in the Con group,the TLR,P13K and Cx43 protein expression levels in the KLA+LY group were lower than those in the KLA group,and the differences were statistically significant(P<0.05);the TLR4 and Cx43 protein expression levels of HCAEC had no statistical difference between the LY group and Con group(P>0.05).Conclusion LY294002 has no significant effect on the expression of TLR4 and Cx43 proteins of HCAEC in the normal culture status.LY294002 can inhibit the high expression of Cx43 in HCAEC induced by KLA.