Alcohol disrupts endoplasmic reticulum function and protein secretion in hepatocytes.
Alcohol disrupts endoplasmic reticulum function and protein secretion in hepatocytes.
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DOI：
10.1111/j.1530-0277.2011.01602.x
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发表时间：
2012-01
期刊：
影响因子：
--
通讯作者：
Sadler KC
中科院分区：
文献类型：
--
作者：
Howarth DL;Vacaru AM;Tsedensodnom O;Mormone E;Nieto N;Costantini LM;Snapp EL;Sadler KC
许多酗酒患者存在血清蛋白缺乏，这是导致他们全身问题的原因之一。未折叠蛋白反应(UPR)是对内质网(ER)蛋白质折叠能力失衡的反应，与肝细胞脂质堆积和细胞凋亡有关，与酒精性肝病相关。我们在体外和体内研究了酒精对肝细胞内质网结构、功能和UPR激活的影响。用50和100 mM乙醇处理表达人细胞色素P450 2E1和小鼠乙醇脱氢酶(VL-17A)的HepG2细胞48h。受精后4d的斑马鱼幼体暴露于350 mM乙醇中32h。用细胞内荧光和斑马鱼的透射电子显微镜观察内质网形态。用定量聚合酶链式反应、原位杂交和Western blotting检测UPR靶基因的活性。通过光漂白后荧光恢复(FRAP)检测细胞内主要内质网伴侣BIP的迁移率。VL-17A细胞可代谢酒精，但乙醇处理后仅有部分UPR靶基因轻微激活。然而，免疫荧光和FRAP检测到内质网碎裂、聚集和未折叠蛋白的聚集，表明尽管缺乏UPR激活，但酒精诱导了一些内质网功能障碍。然而，酒精处理的斑马鱼表现出适度的内质网扩张，并显著诱导了几个UPR靶点。乙醇代谢直接损害肝细胞内质网的结构和功能。斑马鱼是研究酒精性肝病的一种新的体内系统。
Many alcoholic patients have serum protein deficiency that contributes to their systemic problems. The unfolded protein response (UPR) is induced in response to disequilibrium in the protein folding capability of the endoplasmic reticulum (ER) and is implicated in hepatocyte lipid accumulation and apoptosis, which are associated with alcoholic liver disease. We investigated whether alcohol affects ER structure, function and UPR activation in hepatocytes in vitro and in vivo. HepG2 cells expressing human cytochrome P450 2E1 and mouse alcohol dehydrogenase (VL-17A) were treated for up to 48 hours with 50 and 100 mM ethanol. Zebrafish larvae at 4 days post fertilization were exposed to 350 mM ethanol for 32 hours. ER morphology was visualized by fluorescence in cells and transmission electron microscopy in zebrafish. UPR target gene activation was assessed using quantitative PCR, in situ hybridization and Western blotting. Mobility of the major ER chaperone, BIP, was monitored in cells by fluorescence recovery after photobleaching (FRAP). VL-17A cells metabolized alcohol, yet only had slight activation of some UPR target genes following ethanol treatment. However, ER fragmentation, crowding and accumulation of unfolded proteins as detected by immunofluorescence and FRAP, demonstrating that alcohol induced some ER dysfunction despite the lack of UPR activation. Zebrafish treated with alcohol, however, showed modest ER dilation and several UPR targets were significantly induced. Ethanol metabolism directly impairs ER structure and function in hepatocytes. Zebrafish are a novel in vivo system for studying alcoholic liver disease.
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影响因子：
3.8
作者：
Howarth DL;Law SH;Law JM;Mondon JA;Kullman SW;Hinton DE
通讯作者：
Hinton DE
影响因子：
2.9
作者：
Bilotta, J;Barnett, JA;Saszik, S
通讯作者：
Saszik, S
影响因子：
2.7
作者：
Blader, P;Strähle, U
通讯作者：
Strähle, U
影响因子：
16
作者：
Harding, HP;Novoa, I;Ron, D
通讯作者：
Ron, D
影响因子：
2.5
作者：
Dlugos, Cynthia A.;Rabin, Richard A.
通讯作者：
Rabin, Richard A.