HSP-27磷酸化在人臍靜脈內(nèi)皮細(xì)胞凋亡中的作用
發(fā)布時(shí)間:2018-12-14 01:45
【摘要】:目的:研究熱休克蛋白-27(HSP-27)磷酸化在高糖誘導(dǎo)的的人臍靜脈內(nèi)皮細(xì)胞凋亡中的作用;在此基礎(chǔ)上,進(jìn)一步研究HSP-27磷酸化與高糖誘導(dǎo)的人臍靜脈內(nèi)皮細(xì)胞凋亡相關(guān)的信號(hào)轉(zhuǎn)道通路及其可能的機(jī)制。方法:取健康剖腹產(chǎn)孕婦的臍靜脈內(nèi)皮細(xì)胞(HUVECs)進(jìn)行培養(yǎng),選擇2~3代細(xì)胞進(jìn)行實(shí)驗(yàn)。用特異性Phospho-HSP27抗體的蛋白免疫印跡法檢測(cè)HSP-27的活性。采用Annexin-Ⅴ-PI染色流式細(xì)胞技術(shù)檢測(cè)人臍靜脈內(nèi)皮細(xì)胞凋亡。結(jié)果:高糖呈時(shí)間依賴(lài)性誘導(dǎo)人臍靜脈內(nèi)皮細(xì)胞中HSP-27磷酸化,30.5mmol/L高糖作用細(xì)胞24、48、72h,HSP-27磷酸化水平較正常對(duì)照組分別提高了100%(P0.05),182.5%(P0.01),117%(P0.05)。而HSP-27選擇性阻斷劑槲皮素(10umol/L)作用48h可以抑制高糖誘導(dǎo)的HSP-27磷酸化,與高糖組相比槲皮素組HSP-27磷酸化水平降低了52.6%(P0.01),與此同時(shí),可使凋亡增加,內(nèi)皮細(xì)胞凋亡率較高糖組增高了37.6%(P0.01)。高糖誘導(dǎo)的HSP-27磷酸化可以被磷脂酰肌醇-3(phosphoinostide-3kinase,PI3K)選擇性抑制劑LY294002和細(xì)胞外信號(hào)調(diào)節(jié)激酶1/2(extracellular signal regulatedkinase1/2,ERK1/2)選擇性抑制劑U0126分別成濃度依耐性抑制,,與高糖組相比最大抑制率分別為62.6%and56.1%,與此同時(shí),凋亡率上升,分別為50%,120.8%。結(jié)論:高糖可引起HSP-27磷酸化,HSP-27選擇性阻斷劑槲皮素可以抑制高糖誘導(dǎo)的HSP-27磷酸化,并可使內(nèi)皮細(xì)胞凋亡更加明顯,說(shuō)明HSP-27磷酸化對(duì)高糖引起的細(xì)胞凋亡可能起著保護(hù)作用。PI3K/Akt和ERK1/2是高糖誘導(dǎo)的HSP-27磷酸化和細(xì)胞凋亡的重要途徑。
[Abstract]:Aim: to investigate the role of heat shock protein-27 (HSP-27) phosphorylation in high glucose induced apoptosis of human umbilical vein endothelial cells (HUVECs). On this basis, we further studied the signal transduction pathway associated with HSP-27 phosphorylation and apoptosis induced by high glucose in human umbilical vein endothelial cells and its possible mechanism. Methods: umbilical vein endothelial cells (HUVECs) were cultured from healthy pregnant women by caesarean section. The activity of HSP-27 was detected by Western blot with specific Phospho-HSP27 antibody. Apoptosis of human umbilical vein endothelial cells was detected by flow cytometry with Annexin- 鈪
本文編號(hào):2377678
[Abstract]:Aim: to investigate the role of heat shock protein-27 (HSP-27) phosphorylation in high glucose induced apoptosis of human umbilical vein endothelial cells (HUVECs). On this basis, we further studied the signal transduction pathway associated with HSP-27 phosphorylation and apoptosis induced by high glucose in human umbilical vein endothelial cells and its possible mechanism. Methods: umbilical vein endothelial cells (HUVECs) were cultured from healthy pregnant women by caesarean section. The activity of HSP-27 was detected by Western blot with specific Phospho-HSP27 antibody. Apoptosis of human umbilical vein endothelial cells was detected by flow cytometry with Annexin- 鈪
本文編號(hào):2377678
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