發(fā)布時間：2018-04-16 23:38

  本文選題：Kv.蛋白 + 人臍靜脈內(nèi)皮細胞�。� 參考：《華中科技大學學報(醫(yī)學版)》2017年04期

【摘要】：目的研究Kv1.5蛋白在內(nèi)毒素致血管內(nèi)皮細胞損傷中的作用機制。方法體外培養(yǎng)人臍靜脈內(nèi)皮細胞(HUVECs),給予脂多糖(LPS)刺激建立膿毒癥模型,并分為:空白對照組、LPS組、LPS+Kv1.5蛋白通道特異性阻滯劑MT1(250nmol/L)組及LPS+MT2(500nmol/L)組,DAPI染色免疫熒光觀察各組HUVECs凋亡情況,Western blot檢測Caspase-3及Bcl-2蛋白的表達,RT-PCR檢測Caspase-3及Bcl-2基因的表達。結果 DAPI染色免疫熒光顯示,LPS組內(nèi)皮細胞核固縮、凋亡小體形成,凋亡小體形成率由空白對照組的(4.2±0.7)%增加至(26.7±0.8)%(P0.01),而250nmol/L、500nmol/L MT預處理后,凋亡小體形成率從LPS組(26.7±0.8)%分別下降至(12.4±1.0)%、(8.5±0.9)%(均P0.01);Western blot檢測結果顯示,LPS組較空白對照組Bcl-2蛋白表達減少、Caspase-3蛋白表達增加(均P0.01),給予500nmol/L MT預處理后,與LPS組比較,Bcl-2蛋白表達上調、Caspase-3蛋白表達下調(均P0.01);RT-PCR檢測結果顯示,LPS組較空白對照組Bcl-2mRNA表達下調、Caspase-3mRNA表達上調(均P0.01),給予500nmol/L MT預處理后,與LPS組比較,Bcl-2mRNA表達上調、Caspase-3mRNA表達下調(均P0.01)。結論Kv1.5介導膿毒癥相關內(nèi)皮細胞損傷可能與線粒體凋亡途徑有關。
[Abstract]:Objective to investigate the role of Kv1.5 protein in endotoxin-induced vascular endothelial cell injury.Methods Human umbilical vein endothelial cells (HUVECs) were cultured in vitro and stimulated by lipopolysaccharide (LPS) to establish septic model.It was also divided into two groups: LPS-specific blocker MT1 (250 nmol / L) and LPS MT2 (500nmol / L) group. HUVECs apoptosis was observed by immunofluorescence staining. The expression of Caspase-3 and Bcl-2 protein was detected by Western blot and the expression of Caspase-3 and Bcl-2 genes were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results the results of DAPI staining showed that the endothelial cells in lipopolysaccharide group were pyknosis and apoptotic body formation. The percentage of apoptotic bodies was increased from 4.2 鹵0.7% in control group to 26.7 鹵0.8% in control group. However, after pretreatment with 250 nmol / L of 500nmol / L MT, the rate of apoptotic corpuscles increased to 26.7 鹵0.8%.The formation rate of apoptotic corpuscles decreased from 26.7 鹵0.8% in LPS group to 12.4 鹵1.0% in control group (P 0.01). The results of Western blot analysis showed that the expression of Caspase-3 protein in LPS group was significantly lower than that in control group (P 0.01), and the expression of Caspase-3 protein was increased after pretreatment with 500nmol/L MT.Compared with LPS group, the expression of Bcl 2 protein was up-regulated and down-regulated (P 0.01). The results of RT-PCR showed that the expression of Caspase-3 in lipopolysaccharide group was lower than that in control group (P 0.01). After pretreatment with 500nmol/L MT, the expression of Bcl-2 mRNA was up-regulated (P 0.01) and the expression of caspase-3 was down-regulated (P 0.01).Conclusion Kv1.5 mediated sepsis-associated endothelial cell damage may be associated with mitochondrial apoptosis.
【作者單位】： 華中科技大學同濟醫(yī)學院附屬普愛醫(yī)院重癥醫(yī)學科;
【基金】：武漢市衛(wèi)生計生委科研基金資助項目(No.wx14c64) 湖北省自然科學基金資助項目(No.ZRY2014001335)
【分類號】：R459.7
，

本文編號：1761114