摘要
目的在体外培养的大鼠胸主动脉平滑肌细胞(VSMCs)模型上,观察钠/氢交换蛋白1(NHE1)选择性抑制剂cariporide对糖基化终末产物(AGEs)诱导VSMCs表型转化的作用并探讨相关机制。方法采用CCK8检测VSMCs细胞增殖,Transwell试验检测细胞迁移情况;BCECF荧光探针检测细胞内pH值及NHE1活性;Western blot检测NHE1、α-SMA、SM22α及OPN蛋白表达。结果NHE1选择性抑制剂cariporide及Rho激酶(ROCK)抑制剂Y-27632可显著拮抗AGEs诱导的VSMCsα-SMA、SM22α蛋白表达降低及OPN蛋白表达的增多,抑制VSMCs转换为合成表型(去分化表型),从而抑制AGEs诱导的VSMCs的增殖和迁移;同时Y-27632可显著抑制AGEs诱导的细胞内pH值升高,NHE1的活性及蛋白表达。结论该研究结果提示NHE1活化在AGEs诱导VSMCs表型转化中可能起重要作用,同时AGEs调节NHE1活化的机制可能与ROCK相关。
Objective To observe the effect of cariporide,a selective inhibitor of Na+/H+exchanger 1(NHE1),on the phenotypic transformation of thoracic aortic vascular smooth muscle cells(VSMCs)in rats induced by advanced glycation end product(AGEs).Methods CCK8 was used to detect the proliferation of VSMCs cells,transwell assay was used to detect the cell migration,BCECF fluorescence probe was used to detect the intracellular pH value and NHE1 activity,and Western blot was used to detect the expression of NHE1,α-SMA,SM22αand OPN proteins.Results Both cariporide,a selective inhibitor of NHE1,and Rho kinase(ROCK)inhibitor Y-27632 significantly inhibited the AGEs-induced decrease in VSMCsα-SMA and SM22αprotein expression and the increase in OPN expression,and inhibited the conversion of VSMCs to synthetic phenotype(dedifferentiation phenotype),thus inhibiting the proliferation and migration of VSMCs induced by AGEs.Simultaneously,Y-27632 significantly inhibited the increase in intracellular pH,NHE1 activity and protein expression induced by AGEs.Conclusion NHE1 may play an important role in the phenotypic transformation of VSMCs induced by AGEs.ROCK pathway may be related to the mechanism of AGEs-induced NHE1 activity.
作者
吴树金
冷一平
鲍美华
李峰
WU Shu-jin;LENG Yi-ping;BAO Mei-hua;LI Feng(Changsha Medical College,Changsha 410219)
出处
《中南药学》
CAS
2020年第2期194-199,共6页
Central South Pharmacy
基金
湖南省教育厅科学研究项目(No.09C155)。
