Hugan Qingzhi medication ameliorates free fatty acid-induced L02 hepatocyte endoplasmic reticulum stress by regulating the activation of PKC-δ.
Hugan Qingzhi medication ameliorates free fatty acid-induced L02 hepatocyte endoplasmic reticulum stress by regulating the activation of PKC-δ.
复制标题
护肝清脂药物通过调节PKC-δ激活改善游离脂肪酸诱导的L02肝细胞内质网应激
DOI：
10.1186/s12906-020-03164-3
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发表时间：
2020-12-11
影响因子：
3.9
通讯作者：
Zhou B
中科院分区：
文献类型：
--
作者：
Yang M;Chen Z;Xiang S;Xia F;Tang W;Yao X;Zhou B
既往研究发现护肝清脂片对非酒精性脂肪性肝病（NAFLD）具有显著的降脂和抗氧化作用。蛋白质组学分析结果证实，HQT可激活和恢复内质网应激（ERS）途径中的多种蛋白质。然而，其机制仍然混乱。本研究旨在探讨黄芪黄芪总皂苷含药血清对肝脏ERS的影响及其机制。用游离脂肪酸（FFA）诱导L02细胞24 h，建立肝ERS模型，用含药血清预处理24 h。采用油红O染色和甘油三酯检测试剂盒评价细胞内脂质蓄积。透射电镜观察内质网的形态学变化。PKC-δ被特异性siRNA沉默。采用Western blot和RT-qPCR检测ERS、钙代谢紊乱、脂肪变性和胰岛素抵抗相关标志物的表达。用荧光分光光度计记录Ca 2+内流的荧光。HQT含药血清能显著降低细胞内TG含量。此外，它还导致L02细胞ERS标志物的表达显著降低，ER结构得到改善。在FFA诱导的L02肝细胞中，PKC-δ被激活为磷酸化的PKC-δ，而这些变化可以被HQT含药血清逆转。通过沉默L02细胞中PKC-δ的表达，可以恢复ER中SERCA 2的表达和活性，下调IP 3R蛋白的表达，维持细胞内钙稳态，从而减轻FFA诱导的ERS及其脂质蓄积和胰岛素抵抗。结果表明，HQT含药血清对FFA诱导的L02肝细胞ERS、脂肪变性和胰岛素抵抗具有保护作用。其作用机制可能与下调PKC-δ的活化，稳定细胞内钙离子有关。在线版本包含补充材料，可通过10.1186/s12906-020-03164-3获得。
Previous studies have found that Hugan Qingzhi tablet (HQT) has significant lipid-lowering and antioxidant effects on non-alcoholic fatty liver disease (NAFLD). Moreover, the results of proteomic analysis confirmed that various proteins in endoplasmic reticulum stress (ERS) pathway were activated and recovered by HQT. However, its mechanism remains confused. The purpose of this study was to explore the effects of HQT-medicated serum on hepatic ERS and its relevant mechanisms. L02 cells were induced by Free Fatty Acid (FFA) for 24 h to establish a model of hepatic ERS and pretreated with the drug-medicated rat serum for 24 h. Accumulation of intracellular lipid was evaluated using Oil Red O staining and Triglyceride detection kit. The morphological changes of ER were observed by TEM. PKC-δ was silenced by specific siRNA. Western blot and RT-qPCR were applied to detect the expression of markers related to ERS, calcium disorder, steatosis and insulin resistance. The fluorescence of Ca2+ influx was recorded using fluorescence spectrophotometer. HQT-medicated serum significantly decreased the intracellular TG content. Furthermore, it caused significant reduction in the expression of ERS markers and an improvement in ER structure of L02 cells. PKC-δ was activated into phosphorylated PKC-δ in FFA-induced L02 hepatocytes while these changes can be reversed by HQT-medicated serum. Silencing PKC-δ in L02 cells can restore the expression and activity of SERCA2 in ER and down-regulate the expression of IP3R protein to maintain intracellular calcium homeostasis, so as to relieve FFA-induced ERS and its lipid accumulation and insulin resistance. The results concluded that HQT-medicated serum exerts protective effects against hepatic ERS, steatosis and insulin resistance in FFA-induced L02 hepatocyte. And its potential mechanism might be down-regulating the activation of PKC-δ and stabilization of intracellular calcium. The online version contains supplementary material available at 10.1186/s12906-020-03164-3.
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影响因子：
82.9
作者：
通讯作者：
--
葡萄种子提取物对化学诱导肝癌的作用的分子表征：体内和体外分析。
影响因子：
4.6
作者：
Hamza AA;Heeba GH;Elwy HM;Murali C;El-Awady R;Amin A
通讯作者：
Amin A
DOI：
10.1515/jbcpp-2013-0147
发表时间：
2014-11-01
影响因子：
--
作者：
Klymenko, Kateryna;Novokhatska, Tetiana;Soloviev, Anatoly
通讯作者：
Soloviev, Anatoly
影响因子：
4
作者：
Lai, Shujie;Li, Yan;Wen, Liangzhi
通讯作者：
Wen, Liangzhi
影响因子：
9
作者：
De Ford C;Heidersdorf B;Haun F;Murillo R;Friedrich T;Borner C;Merfort I
通讯作者：
Merfort I