發(fā)布時(shí)間：2018-10-14 18:37

【摘要】：目的 觀察普通肝素(UFH)對(duì)高遷移率族蛋白1(HMGB 1)介導(dǎo)的人臍靜脈內(nèi)皮細(xì)胞屏障通透性損傷的保護(hù)作用,探討UFH對(duì)HMGB 1介導(dǎo)的胞質(zhì)緊密粘連蛋白-1(ZO-1)表達(dá)缺失的保護(hù)機(jī)制。方法 將人臍靜脈內(nèi)皮細(xì)胞進(jìn)行體外培養(yǎng),人臍靜脈內(nèi)皮細(xì)胞株傳代培養(yǎng)后分為4組(n=5):空白對(duì)照組(加入等量PBS)、HMGB 1處理組(100 ng/ml)、UFH對(duì)照組(UFH 10 U/ml)、HMGB 1及UFH處理組(100 ng/ml HMGB 1+UFH 10 U/ml)。MTT法測(cè)定內(nèi)皮細(xì)胞存活率,用Transwell小室法測(cè)定單層內(nèi)皮屏障通透性,用免疫熒光染色法測(cè)定ZO-1的表達(dá)分布,蛋白免疫印跡(Western blot)法檢測(cè)ZO-1蛋白及核因子-κB(NF-κB)的表達(dá)。結(jié)果 HMGB 1(100 ng/ml)對(duì)內(nèi)皮細(xì)胞活性無(wú)抑制作用(P0.05)。UFH預(yù)處理后可減少HMGB1所致的內(nèi)皮細(xì)胞通透性增加(P0.05)。UFH預(yù)處理后可降低HMGB 1所致內(nèi)皮細(xì)胞ZO-1密閉環(huán)的減少和破壞,使ZO-1熒光強(qiáng)度增強(qiáng),ZO-1蛋白表達(dá)增加,并使NF-κB的核易位減少。結(jié)論 UFH可保護(hù)HMGB 1介導(dǎo)的內(nèi)皮細(xì)胞ZO-1的表達(dá)缺失,進(jìn)而改善內(nèi)皮細(xì)胞屏障通透性,其機(jī)制與減少NF-κB核易位相關(guān)。
[Abstract]:Objective to investigate the protective effect of common heparin (UFH) on barrier permeability injury of human umbilical vein endothelial cells mediated by high mobility group protein 1 (HMGB 1), and to explore the protective mechanism of UFH against HMGB 1 mediated loss of cytoplasmic compact adhesion protein-1 (ZO-1) expression. Methods Human umbilical vein endothelial cells were cultured in vitro. Human umbilical vein endothelial cells (HUVEC) were divided into 4 groups after subculture: blank control group (UFH 10 U/ml), HMGB 1) and UFH treated group (100 ng/ml HMGB 1 UFH 10 U/ml). MTT). The permeability of endothelial barrier was measured by Transwell chamber method, the expression of ZO-1 was detected by immunofluorescence staining, and the expression of ZO-1 protein and nuclear factor- 魏 B (NF- 魏 B) were detected by Western blot (Western blot). Results HMGB 1 (100 ng/ml) had no inhibitory effect on endothelial cell activity (P0.05). UFH pretreatment decreased the increase of endothelial cell permeability induced by HMGB1 (P0.05). UFH pretreatment decreased the decrease and destruction of ZO-1 closed loop induced by HMGB 1. The fluorescence intensity of ZO-1 was increased, the expression of ZO-1 protein was increased, and the nuclear translocation of NF- 魏 B was decreased. Conclusion UFH can protect HMGB 1 mediated ZO-1 expression deletion in endothelial cells and improve the permeability of endothelial cell barrier. The mechanism is related to the reduction of NF- 魏 B translocation.
【作者單位】： 中國(guó)醫(yī)科大學(xué)附屬第一醫(yī)院重癥醫(yī)學(xué)科;

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