【摘要】：目的探討小鼠肺微血管內(nèi)皮細(xì)胞(mPMVECs)熱休克蛋白A12B(HSPA12B)表達(dá)下調(diào)對脂多糖(LPS)誘導(dǎo)的細(xì)胞炎癥反應(yīng)、遷移及超微結(jié)構(gòu)改變的影響。方法體外傳代培養(yǎng)的mPMVECs分為LPS組(添加1μg/mL LPS)、LPS+siRNA組[瞬時轉(zhuǎn)染法將HSPA12B相應(yīng)基因片段干擾小RNA(siRNA)寡核苷酸轉(zhuǎn)入mPMVECs中,再加入1μg/mL LPS)]和LPS+NC組[瞬時轉(zhuǎn)染法將平行對照(NC)siRNA寡核苷酸轉(zhuǎn)入mPMVECs中,再加入1μg/mL LPS]。對LPS+siRNA組和LPS+NC組,分別采用Transwell試驗和細(xì)胞劃痕實驗觀察細(xì)胞遷移情況;應(yīng)用透射電子顯微鏡(透射電鏡)觀察mPMVECs超微結(jié)構(gòu)改變;應(yīng)用Real-Time PCR檢測各組細(xì)胞中腫瘤壞死因子-α(TNF-α)、白介素-6(IL-6)、IL-10和IL-1βmRNA的表達(dá)。結(jié)果與LPS+NC組比較,LPS+siRNA組mPMVECs的線粒體損傷加重,內(nèi)質(zhì)網(wǎng)水腫更明顯;細(xì)胞內(nèi)TNF-α、IL-6和IL-10mRNA的表達(dá)顯著上調(diào)(P0.05),而IL-1β表達(dá)下調(diào),但差異無統(tǒng)計學(xué)意義(P0.05);細(xì)胞遷移功能顯著下降(P0.05)。結(jié)論下調(diào)HSPA12B表達(dá)后,mPMVECs經(jīng)LPS刺激的炎癥反應(yīng)增強(qiáng),細(xì)胞遷移功能下降。HSPA12B可能參與保護(hù)mPMVECs免受LPS侵襲的過程。
[Abstract]:Objective to investigate the effect of down-regulation of (mPMVECs) heat shock protein A12B (HSPA12B) expression in mouse pulmonary microvascular endothelial cells on cellular inflammation, migration and ultrastructural changes induced by lipopolysaccharide (LPS). Methods mPMVECs cultured in vitro was divided into LPS group (1 渭 g/mL LPS) and LPS siRNA group [transient transfection of HSPA12B gene fragment interfering small RNA (siRNA) oligonucleotide into mPMVECs. In the LPS NC group, parallel control (NC) siRNA oligonucleotides were transferred into mPMVECs by transient transfection and then 1 渭 g/mL LPS was added. In LPS siRNA group and LPS NC group, cell migration was observed by Transwell test and cell scratch test, and ultrastructural changes of mPMVECs were observed by transmission electron microscope (TEM). The expression of tumor necrosis factor- 偽 (TNF- 偽), interleukin-6 (IL-6) IL-10 and IL-1 尾 mRNA were detected by Real-Time PCR. Results compared with LPS NC group, the mitochondria damage and endoplasmic reticulum edema were more serious in mPMVECs group than those in LPS NC group, the expression of TNF- 偽 IL-6 and IL-10mRNA was significantly up-regulated (P0.05), but the expression of IL-1 尾 was down-regulated, but the difference was not statistically significant (P0.05), and the cell migration function was significantly decreased (P0.05). Conclusion after down-regulation of HSPA12B expression, the inflammatory response of mPMVECs stimulated by LPS is enhanced, and the cell migration function is decreased. HSPA12B may be involved in the process of protecting mPMVECs from LPS invasion.
【作者單位】： 第二軍醫(yī)大學(xué)長海醫(yī)院麻醉科;上海交通大學(xué)醫(yī)學(xué)院附屬第三人民醫(yī)院麻醉科;
【分類號】：R563
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本文編號：2156856