Obeticholic acid attenuates human immunodeficiency virus/alcohol metabolism-induced pro-fibrotic activation in liver cells.
Obeticholic acid attenuates human immunodeficiency virus/alcohol metabolism-induced pro-fibrotic activation in liver cells.
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奥贝胆酸减弱人类免疫缺陷病毒/酒精代谢诱导的肝细胞促纤维化活化。
DOI：
10.4254/wjh.v12.i11.965
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发表时间：
2020-11-27
影响因子：
2.4
通讯作者：
Osna NA
中科院分区：
文献类型：
--
作者：
New-Aaron M;Ganesan M;Dagur RS;Kharbanda KK;Poluektova LY;Osna NA
人类免疫缺陷病毒（HIV）感染的发病率和死亡率通常与肝脏疾病相关，肝脏疾病缓慢进展为严重的肝功能障碍。有多种损害加剧了HIV相关的肝损伤，包括HIV相关的脂质代谢和脂肪周转失调，嗜肝病毒的合并感染和酒精滥用。如我们之前所报道的，肝细胞暴露于HIV和酒精代谢产物会导致高氧化应激，损害蛋白酶体和溶酶体功能，导致HIV在这些细胞中积累，最终导致细胞凋亡并最终促进肝纤维化的发展。研究奥贝胆酸（OCA）是否可预防HIV/乙醇代谢诱导的肝毒性以及随后由HIV+凋亡肝细胞吞噬引起的肝星状细胞（HSC）活化。 在存在或不存在OCA的情况下，将Huh7.5-HCV（RLW）细胞暴露于HIV和乙酰丙酮脱氢系统（AGS）。在细胞中，我们检测了HIV相关标志物的表达：HIVgagRNA-通过实时聚合酶链反应（PCR），p24-通过western blot，HIV DNA-通过半巢式PCR，整合HIV DNA-通过ddPCR。溶酶体和蛋白酶体的活动进行了测量，使用荧光标记的底物。对于肝细胞凋亡，通过蛋白质印迹法观察裂解的半胱天冬酶3和裂解的PARP，通过M30 ELISA观察上清液中的细胞角蛋白18。从暴露于UV光的未经处理和经HIV处理的RLW细胞产生凋亡体。HSC的促纤维化激活通过Col 1A 1和转化生长因子-β mRNA表达来表征，而炎性小体激活通过NLRP 3、caspase 1、白细胞介素（IL）-6、IL-1β mRNA表达来表征。在RLW细胞中，OCA处理减弱了HIV-AGS诱导的HIVgagRNA、HIV DNA和p24积累。OCA抑制活性氧的产生并恢复胰凝乳蛋白酶样蛋白酶体活性以及组织蛋白酶B溶酶体活性。OCA还减少了RLW细胞中HIV-AGS触发的凋亡。暴露于HSC中的HIV凋亡肝细胞阻止了炎性小体的激活，并诱导了这些细胞的促纤维化激活。 我们的结论是，通过抑制氧化应激和恢复蛋白酶体和溶酶体功能受损的艾滋病毒和乙醇代谢，OCA减少艾滋病毒在肝细胞中的积累，导致下调这些细胞的凋亡。此外，OCA逆转由吞噬含HIV的凋亡肝细胞触发的HSC的促纤维化和炎性小体相关活化，可能有助于抑制肝纤维化发展。
The morbidity and mortality of human immunodeficiency virus (HIV)-infection is often associated with liver disease, which progresses slowly into severe liver dysfunction. There are multiple insults which exacerbate HIV-related liver injury, including HIV-associated dysregulation of lipid metabolism and fat turnover, co-infections with hepatotropic viruses and alcohol abuse. As we reported before, exposure of hepatocytes to HIV and alcohol metabolites causes high oxidative stress, impairs proteasomal and lysosomal functions leading to accumulation of HIV in these cells, which end-ups with apoptotic cell death and finally promotes development of liver fibrosis. To study whether obeticholic acid (OCA) prevents HIV/ethanol metabolism-induced hepatotoxicity and subsequent activation of hepatic stellate cells (HSC) by HIV+ apoptotic hepatocyte engulfment. Huh7.5-CYP (RLW) cells were exposed to HIV and acetaldehyde-generating system (AGS) in the presence or absence of OCA. In the cells, we measured the expression of HIV-related markers: HIVgagRNA-by real-time polymerase chain reaction (PCR), p24- by western blot, HIV DNA-by semi-nested PCR, integrated HIV DNA-by ddPCR. Lysosomal and proteasomal activities were measured using fluorometrically-labeled substrates. For hepatocyte apoptosis, cleaved caspase 3 and cleaved PARP were visualized by western blot and cytokeratin 18- by M30 ELISA-in supernatants. Apoptotic bodies were generated from untreated and HIV-treated RLW cells exposed to UV light. Pro-fibrotic activation of HSC was characterized by Col1A1 and transforming growth factor-β mRNAs, while inflammasome activation- by NLRP3, caspase 1, interleukin (IL)-6, IL-1β mRNA levels. In RLW cells, OCA treatment attenuated HIV-AGS-induced accumulation of HIVgagRNA, HIV DNA and p24. OCA suppressed reactive oxygen species production and restored chymotrypsin-like proteasome activity as well as cathepsin B lysosome activity. OCA also decreased HIV-AGS-triggered apoptosis in RLW cells. Exposure of HIV-containing apoptotic hepatocytes to HSC prevented activation of inflammasome and induced pro-fibrotic activation in these cells. We conclude that by suppressing oxidative stress and restoring proteasomal and lysosomal functions impaired by HIV and ethanol metabolism, OCA decreases accumulation of HIV in hepatocytes, leading to down-regulation of apoptosis in these cells. In addition, OCA reverses pro-fibrotic and inflammasome-related activation of HSC triggered by engulfment of HIV-containing apoptotic hepatocytes, potentially contributing to suppression of liver fibrosis development.
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DOI：
10.1016/j.jcmgh.2017.10.004
发表时间：
2018
影响因子：
7.2
作者：
Ganesan M;Tikhanovich I;Vangimalla SS;Dagur RS;Wang W;Poluektova LI;Sun Y;Mercer DF;Tuma D;Weinman SA;Kharbanda KK;Osna NA
通讯作者：
Osna NA
影响因子：
5.5
作者：
Ganesan, Murali;New-Aaron, Moses;Osna, Natalia A.
通讯作者：
Osna, Natalia A.
影响因子：
2.9
作者：
Sterling, Richard K.;Smith, Paula G.;Brunt, Elizabeth M.
通讯作者：
Brunt, Elizabeth M.
影响因子：
3.7
作者：
Renga, Barbara;Francisci, Daniela;Fiorucci, Stefano
通讯作者：
Fiorucci, Stefano
影响因子：
5.1
作者：
Iracheta-Vellve A;Calenda CD;Petrasek J;Ambade A;Kodys K;Adorini L;Szabo G
通讯作者：
Szabo G