摘要
目的探讨缝隙连接(GJ)是否通过PI3K/Akt信号途径影响氧化低密度脂蛋白(Ox LDL)诱导的大鼠胸大动脉平滑肌细胞(A7r5)增殖。方法培养A7r5细胞株,将细胞分4组,即对照组、Ox LDL组、Ox LDL+二甲基亚砜(DMSO)组和Ox LDL+Akt抑制剂(MK-2206)组。用MTS法及流式细胞仪检测A7r5的增殖活性;流式细胞仪检测各组发绿色荧光的供体细胞(Q3)与双阴性受体细胞(Q4)的比值(Q3/Q4)变化,从而分析GJ功能改变;Western blot法检测细胞中Cx43、Akt、p-Akt蛋白表达。结果与对照组比较,Ox LDL组及Ox LDL+DMSO组A7r5细胞增殖活性增加,细胞周期S期细胞比率增高(P<0.05);与Ox LDL组比较,Ox LDL+MK-2206组两者均降低(P<0.05)。与对照组相比较,Ox LDL组及Ox LDL+DMSO组A7r5细胞的Q3/Q4比值增大,GJ功能增强(P<0.05);与Ox LDL组比较,Ox LDL+MK-2206组Q3/Q4比值减小,GJ功能显著降低(P<0.05)。与对照组相比较,Ox LDL组和Ox LDL+DMSO组A7r5细胞的Cx43、p-Akt蛋白表达增强(P<0.05);与Ox LDL组比较,Ox LDL+MK-2206组Cx43、p-Akt蛋白表达减弱(P<0.05)。与对照组比较,Ox LDL组、LDL+DMSO组及Ox LDL+MK-2206组A7r5细胞的Akt蛋白均变化不明显。结论 GJ可以通过PI3K/Akt信号途径影响Dx LDL诱导的A7r5细胞增殖。
Objective To study whether gap junctions affect A7r5 cell proliferation mediated by OxLDL through the PI3K/Akt signaling pathway. Methods The A7r5 cell line was cultured,then the cells were divided into four groups：control group, OxLDL group, OxLDL plus DMSO group and OxLDL plus MK -2206 group. The cellular proliferation activity was measured by MTS and Flow cytome- try. Flow cytometry was used to detect the ratio of Q3 （Calcein labeled cells） to Q4 （double negative cells） for the analysis of the chan- ges of the gap junction function. The expression of connexins 43 （ Cx43 ）, Akt and p - Akt were detected by western blot. Results The ATr5 cell proliferation activity and the S - phase fraction detected in the OxLDL group and the OxLDL plus DMSO group were increased obviously compared with those in the control group （P 〈 0.05 ）. The A7r5 cell proliferation activity and the S - phase fraction in the Ox- LDL plus MK - 2206 group were decreased compared with those in the OxLDL group （P 〈 O. 05）. The Q3/Q4 ratio in the OxLDL group and the OxLDL plus DMSO group was significantly higher than that in the control group （ P 〈 0.05 ）, while the Q3/Q4 ratio in the Ox- LDL plus MK - 2206 group was lower （P 〈 0.05） than that in the OxLDL group （P 〈 O. 05）. The expression of Cx43 and p - Akt pro- teins in the OxLDL group and OxLDL ＋ DMSO group were increased compared with the control group （ P 〈 0.05 ）. Compared with the OxLDL group,the expression of Cx43 and p - Akt proteins in the OxLDL plus MK - 2206 group were significantly decreased （ P 〈 0.05 ）. Compared with the control group, the expression of Akt proteins in the other three groups change is not obviously. Conclusion Gap junctions could affect A7r5 cell proliferation mediated by OxLDL through the PI3K/Akt signaling pathway.
出处
《宁夏医学杂志》
CAS
2015年第12期1068-1070,共3页
Ningxia Medical Journal
基金
宁夏自然科学基金资助项目(NZ13187)
