本文關(guān)鍵詞： ROBO ARF 通透性 基因過表達(dá) 糖尿病腎病　出處：《解放軍醫(yī)學(xué)雜志》2017年03期 　論文類型：期刊論文

【摘要】：目的探討ROBO4基因過表達(dá)對(duì)高糖誘導(dǎo)的人腎小球內(nèi)皮細(xì)胞(HRGEC)高通透性的影響。方法體外培養(yǎng)HRGEC,以攜帶ROBO4基因的重組慢病毒載體感染HRGEC獲得過表達(dá)ROBO4的HRGEC。將細(xì)胞分為低糖培養(yǎng)組和高糖培養(yǎng)組,采用Western blotting檢測(cè)ROBO4和ARF6蛋白的表達(dá),異硫氰酸熒光素右旋糖酐(FITC-Dextran)法檢測(cè)HRGEC的通透性,CCK-8法測(cè)定細(xì)胞存活率。結(jié)果慢病毒在HRGEC中的72h轉(zhuǎn)染率達(dá)80%,與空載體組相比ROBO4轉(zhuǎn)染組HRGEC細(xì)胞的ROBO4蛋白明顯過表達(dá)(P0.05),CCK8檢測(cè)顯示慢病毒轉(zhuǎn)染后HRGEC細(xì)胞活性不受影響。與低糖組相比,高糖組ROBO4蛋白表達(dá)在12h時(shí)點(diǎn)明顯增高(P0.05),24h回落,72h下降最為明顯(P0.05),而ARF6蛋白的表達(dá)在12h開始明顯升高,72h時(shí)升高最為明顯(P0.05)。高糖組FITC-Dextran層HRGEC屏障的通透性在24h開始逐步升高,72h時(shí)升高最為明顯(P0.05);與空載體組相比,ROBO4過表達(dá)可顯著抑制高糖條件下ARF6的表達(dá)及降低體外單層HRGEC屏障的FITC-Dextran通透性。結(jié)論 ROBO4過表達(dá)可降低高糖條件下HRGEC的通透性,激活ROBO4有望成為治療糖尿病腎病內(nèi)皮功能障礙的新方向。
[Abstract]:Objective to investigate the effect of overexpression of ROBO4 gene on high permeability of human glomerular endothelial cells induced by high glucose. Methods HRGEC was cultured in vitro. The recombinant lentivirus vector carrying ROBO4 gene was used to infect HRGEC. The cells were divided into low glucose culture group and high glucose culture group. The expression of ROBO4 and ARF6 protein was detected by Western blotting. Fluorescein isothiocyanate (FITC-Dextranan) was used to detect the permeability of HRGEC. Results the transfection rate of lentivirus in HRGEC for 72 hours was 80%. Compared with the empty vector group, the ROBO4 protein of HRGEC cells transfected with ROBO4 was significantly overexpressed (P0.05). CCK8 analysis showed that the activity of HRGEC cells was not affected after lentivirus transfection. Compared with low glucose group, the expression of ROBO4 protein in high glucose group was significantly higher than that in low glucose group at 12h (P 0.05). The expression of ARF6 protein began to increase at 12 h. The permeability of HRGEC barrier in FITC-Dextran layer of high glucose group increased gradually at 24 h. At 72 h, the increase of P0.05 was the most obvious. Compared with empty carrier group. Overexpression of ROBO4 could significantly inhibit the expression of ARF6 and reduce the FITC-Dextran permeability of monolayer HRGEC barrier in vitro. Overexpression of ROBO4 can reduce the permeability of HRGEC under high glucose condition. Activation of ROBO4 is expected to be a new direction in the treatment of diabetic nephropathy endothelial dysfunction.
【作者單位】： 第三軍醫(yī)大學(xué)新橋醫(yī)院腎內(nèi)科;
【基金】：國(guó)家自然科學(xué)基金(81370821) 重慶市自然科學(xué)基金重點(diǎn)項(xiàng)目(cstc2013jjB10023)~~
【分類號(hào)】：R692.9;R587.2
【正文快照】： 糖尿病發(fā)病率逐年上升,糖尿病腎病(diabeticnephropathy,DN)仍然是臨床難以攻克的慢性疾病之一[1]。近年研究發(fā)現(xiàn),腎小球高通透性(endothelial/glomerular hyperpermeability)可能是糖尿病視網(wǎng)膜病、糖尿病系統(tǒng)血管病變和DN微量白蛋白尿啟動(dòng)的重要原因。已知高糖、晚期糖基化

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