PNPLA3 downregulation exacerbates the fibrotic response in human hepatic stellate cells.
PNPLA3 downregulation exacerbates the fibrotic response in human hepatic stellate cells.
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DOI：
10.1371/journal.pone.0260721
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发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Pocai A
中科院分区：
文献类型：
--
作者：
Rady B;Nishio T;Dhar D;Liu X;Erion M;Kisseleva T;Brenner DA;Pocai A
非酒精性脂肪性肝炎 (NASH) 部分是由代谢紊乱与易感基因变异相互作用导致的，导致肝脏相关并发症和死亡。最强的遗传决定因素是含有 patatin 样磷脂酶结构域的蛋白 3 (PNPLA3 p.I148M) 中高度普遍的错义变体。在人肝脏肝细胞中，PNPLA3 定位于脂滴表面，据信突变形式可增强脂质积累和促炎细胞因子的释放。关于 PNPLA3 在肝星状细胞 (HSC) 中的作用知之甚少。在这里，我们表征了从携带野生型（n = 8 C/C）和杂合子（n = 6，C/G）或纯合子（n = 6，G/G）PNPLA3 I148M的患者获得的HSC，并研究了基因型和PNPLA3下调对基线和TGF-β刺激的纤维化基因表达的影响。所有基因型的 HSC 均显示出相当的 PNPLA3 基线水平以及纤维化基因 α-SMA、COL1A1、TIMP1 和 SMAD7 的表达。 TGF-β治疗增加了所有3种基因型中PNPLA3的表达（约2倍），并对几种纤维形成基因的表达产生了类似的刺激。在携带野生型 (WT) PNPLA3 的原代人 HSC 中，siRNA 处理使 PNPLA3 mRNA 减少 79%，导致用 TGF-β 刺激的细胞中 α-SMA、Col1a1、TIMP1 和 SMAD7 的表达增加。同样，在携带 C/G 或 G/G 基因型的 HSC 中敲除 PNPLA3 会导致 TGF-β 诱导的纤维化基因表达增强。在没有 TGF-β 治疗的情况下，PNPLA3 的敲低不会影响纤维化基因的表达。总之，这些数据表明 HSC 中 I148M PNPLA3 突变的存在对基线激活没有影响，并且无论基因型如何，PNPLA3 的下调都会加剧纤维化反应。
Non-alcoholic steatohepatitis (NASH) results, in part, from the interaction of metabolic derangements with predisposing genetic variants, leading to liver-related complications and mortality. The strongest genetic determinant is a highly prevalent missense variant in patatin-like phospholipase domain-containing protein 3 (PNPLA3 p.I148M). In human liver hepatocytes PNPLA3 localizes to the surface of lipid droplets where the mutant form is believed to enhance lipid accumulation and release of pro-inflammatory cytokines. Less is known about the role of PNPLA3 in hepatic stellate cells (HSCs). Here we characterized HSC obtained from patients carrying the wild type (n = 8 C/C) and the heterozygous (n = 6, C/G) or homozygous (n = 6, G/G) PNPLA3 I148M and investigated the effect of genotype and PNPLA3 downregulation on baseline and TGF-β-stimulated fibrotic gene expression. HSCs from all genotypes showed comparable baseline levels of PNPLA3 and expression of the fibrotic genes α-SMA, COL1A1, TIMP1 and SMAD7. Treatment with TGF-β increased PNPLA3 expression in all 3 genotypes (~2-fold) and resulted in similar stimulation of the expression of several fibrogenic genes. In primary human HSCs carrying wild-type (WT) PNPLA3, siRNA treatment reduced PNPLA3 mRNA by 79% resulting in increased expression of α-SMA, Col1a1, TIMP1, and SMAD7 in cells stimulated with TGF-β. Similarly, knock-down of PNPLA3 in HSCs carrying either C/G or G/G genotypes resulted in potentiation of TGF-β induced expression of fibrotic genes. Knockdown of PNPLA3 did not impact fibrotic gene expression in the absence of TGF-β treatment. Together, these data indicate that the presence of the I148M PNPLA3 mutation in HSC has no effect on baseline activation and that downregulation of PNPLA3 exacerbates the fibrotic response irrespective of the genotype.
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影响因子：
6
作者：
Schwartz BE;Rajagopal V;Smith C;Cohick E;Whissell G;Gamboa M;Pai R;Sigova A;Grossman I;Bumcrot D;Sasidharan K;Romeo S;Sehgal A;Pingitore P
通讯作者：
Pingitore P
影响因子：
5.1
作者：
Bruschi, Francesca Virginia;Claudel, Thierry;Trauner, Michael
通讯作者：
Trauner, Michael
影响因子：
13.5
作者：
Fuchs, Claudia D.;Claudel, Thierry;Trauner, Michael
通讯作者：
Trauner, Michael
影响因子：
13.5
作者：
Bruschi, Francesca Virginia;Claudel, Thierry;Trauner, Michael
通讯作者：
Trauner, Michael
影响因子：
8.3
作者：
Sandhu, Bynvant;Matos, Maria C. Perez;Jiang, Z. Gordon
通讯作者：
Jiang, Z. Gordon