本文選題：喹乙醇 + 內(nèi)質(zhì)網(wǎng)應(yīng)激�。� 參考：《衛(wèi)生研究》2015年03期

【摘要】：目的通過(guò)喹乙醇染毒人源腎小管上皮細(xì)胞(HK-2細(xì)胞)并檢測(cè)活性氧(ROS)和凋亡相關(guān)蛋白的表達(dá),探討喹乙醇腎臟毒性產(chǎn)生過(guò)程及其可能的內(nèi)質(zhì)網(wǎng)應(yīng)激相關(guān)凋亡機(jī)制。方法分別以不同濃度(1、2、3、4、5、6、7和8μmol/ml)喹乙醇染毒HK-2細(xì)胞24 h,噻唑藍(lán)(MTT)比色法檢測(cè)細(xì)胞增殖率以確定劑量-效應(yīng)關(guān)系;Hoechst-33258熒光染色檢測(cè)各組細(xì)胞凋亡形態(tài),流式細(xì)胞儀檢測(cè)各組細(xì)胞凋亡率和細(xì)胞內(nèi)活性氧含量;免疫印跡法(Western blot)檢測(cè)內(nèi)質(zhì)網(wǎng)應(yīng)激相關(guān)凋亡蛋白葡萄糖調(diào)節(jié)蛋白78(GRP78)、葡萄糖調(diào)節(jié)蛋白94(GRP94)和凋亡促進(jìn)因子CCAAT增強(qiáng)子結(jié)合蛋白同源蛋白(CHOP)的表達(dá)。結(jié)果根據(jù)MTT毒性實(shí)驗(yàn)結(jié)果 ,喹乙醇對(duì)HK-2細(xì)胞凋亡效應(yīng)的適宜劑量確定為1、2、3和4μmol/ml。在喹乙醇不同劑量觀察組中,喹乙醇染毒2μmol/ml以上劑量組,細(xì)胞凋亡率、內(nèi)質(zhì)網(wǎng)凋亡相關(guān)蛋白GRP79、GRP94和CHOP表達(dá)增加,喹乙醇各染毒劑量均可見(jiàn)活性氧水平增加(P0.05);在喹乙醇不同染毒時(shí)間觀察組中,喹乙醇染毒12和24 h組,細(xì)胞凋亡率、內(nèi)質(zhì)網(wǎng)凋亡相關(guān)蛋白GRP79、GRP94表達(dá)增加,喹乙醇染毒6、12和24 h組,活性氧水平、內(nèi)質(zhì)網(wǎng)凋亡相關(guān)蛋白CHOP表達(dá)增加(P0.05)。結(jié)論喹乙醇可致腎小管上皮細(xì)胞發(fā)生細(xì)胞凋亡從而造成腎臟毒性,凋亡的發(fā)生可能與內(nèi)質(zhì)網(wǎng)應(yīng)激相關(guān)凋亡途徑相關(guān)。
[Abstract]:Objective to study the expression of reactive oxygen species (ROS) and apoptosis related protein in human renal tubular epithelial cells (HK-2 cells) and to explore the mechanism of renal toxicity induced by olq and its possible mechanism of apoptosis related to endoplasmic reticulum stress. Methods HK-2 cells with different concentrations (1,2,3,4,5,6,7 and 8 mol/ml) were infected with 24 h, thiazole, respectively. Blue (MTT) colorimetric assay was used to determine the cell proliferation rate to determine the dose effect relationship; Hoechst-33258 fluorescence staining was used to detect the apoptosis morphology of each group. Flow cytometry was used to detect the apoptosis rate and intracellular reactive oxygen content of each group; Western blot (Western blot) was used to detect the endoplasmic reticulum associated apoptosis protein glucose regulator protein 78 (GRP78), and the grape The expression of sugar regulated protein 94 (GRP94) and apoptosis promoting factor CCAAT enhancer binding protein homologous protein (CHOP). Results according to the results of MTT toxicity test, the optimum dosage of ethanol for the apoptosis of HK-2 cells was determined to be 1,2,3 and 4 mol/ml. in the observation group of different doses of oloethanol. The dose group over 2 mu mol/ml by olq, and the cell apoptosis Rate, the expression of GRP79, GRP94 and CHOP of endoplasmic reticulum increased, and the level of reactive oxygen species increased (P0.05) in all doses of ethyl alcohol. In the group of 12 and 24 h, the rate of apoptosis, the apoptosis related to the endoplasmic reticulum, the expression of egg white GRP79, the expression of GRP94, and the activity of 6,12 and 24 h in ethyl alcohol. The expression of apoptosis related protein CHOP in endoplasmic reticulum increased (P0.05). Conclusion ethanol can induce apoptosis in renal tubular epithelial cells and induce renal toxicity. Apoptosis may be related to the apoptosis pathway associated with endoplasmic reticulum stress.
【作者單位】： 中國(guó)疾病預(yù)防控制中心職業(yè)衛(wèi)生與中毒控制所毒理室 中國(guó)疾病預(yù)防控制中心化學(xué)污染物與健康重點(diǎn)實(shí)驗(yàn)室;
【基金】：環(huán)保公益性行業(yè)科研專項(xiàng)(No.201109038)
【分類號(hào)】：R114

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 張婷;陳倩;湯樹生;靳溪;鄒家杰;劉鳳英;張q，

本文編號(hào)：2016636