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BMC Complementary Medicine and Therapies
Published in: BMC Complementary Medicine and Therapies 1/2020

01-12-2020 | Fenofibrate | Research article

Hugan Qingzhi medication ameliorates free fatty acid-induced L02 hepatocyte endoplasmic reticulum stress by regulating the activation of PKC-δ

Authors: Miaoting Yang, Zhijuan Chen, Shijian Xiang, Fan Xia, Waijiao Tang, Xiaorui Yao, Benjie Zhou

Published in: BMC Complementary Medicine and Therapies | Issue 1/2020

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Abstract

Background

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.

Methods

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.

Results

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.

Conclusions

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.
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Literature
1.
2.
Day CP, James OFW. Steatohepatitis: a tale of two “hits”? Gastroenterology. 1998;114(4):842–5.PubMedCrossRef
3.
Gentile CL, Pagliassotti MJ. The role of fatty acids in the development and progression of nonalcoholic fatty liver disease. J Nutr Biochem. 2008;19(9):567–76.PubMedPubMedCentralCrossRef
4.
5.
6.
Ji C. Dissection of endoplasmic reticulum stress signaling in alcoholic and non-alcoholic liver injury. J Gastroenterol Hepatol. 2008;23(Suppl 1):S16–24.PubMedPubMedCentralCrossRef
7.
Afrin R, Arumugam S, Wahed MII, Pitchaimani V, Karuppagounder V, Sreedhar R, Harima M, Suzuki H, Miyashita S, Nakamura T, et al. Attenuation of endoplasmic reticulum stress-mediated liver damage by mulberry leaf diet in Streptozotocin-induced diabetic rats. Am J Chin Med. 2016;44(1):87.PubMedCrossRef
8.
Newton AC. Protein kinase C: poised to signal. Am J Physiol Endoc Metab. 2010;298(3):E395–402.CrossRef
9.
Bezy O, Tran TT, Pihlajamäki J, Suzuki R, Emanuelli B, Winnay J, Mori MA, Haas J, Biddinger SB, Leitges M, et al. PKCδ regulates hepatic insulin sensitivity and hepatosteatosis in mice and humans. J Clin Invest. 2011;121(6):2504–17.PubMedPubMedCentralCrossRef
10.
Greene MW, Burrington CM, Luo Y, Ruhoff MS, Lynch DT, Chaithongdi N. PKCδ is activated in the liver of obese Zucker rats and mediates diet-induced whole body insulin resistance and hepatocyte cellular insulin resistance. J Nutr Biochem. 2014;25(3):281–8.PubMedCrossRef
11.
Greene MW, Burrington CM, Lynch DT, Davenport SK, Johnson AK, Horsman MJ, Chowdhry S, Zhang J, Sparks JD, Tirrell PC. Lipid metabolism, oxidative stress and cell death are regulated by PKC delta in a dietary model of nonalcoholic steatohepatitis. PLoS One. 2014;9(1):e85848.PubMedPubMedCentralCrossRef
12.
Greene MW, Burrington CM, Ruhoff MS, Johnson AK, Chongkrairatanakul T, Kangwanpornsiri A. PKCδ is activated in a dietary model of Steatohepatitis and regulates endoplasmic reticulum stress and cell death. J Biol Chem. 2010;285(53):42115–29.PubMedPubMedCentralCrossRef
13.
Hamza AA, Heeba GH, Elwy HM, Murali C, El-Awady R, Amin A. Molecular characterization of the grape seeds extract's effect against chemically induced liver cancer: in vivo and in vitro analyses. Sci Rep. 2018;8(1):1270.PubMedPubMedCentralCrossRef
14.
Al-Dabbagh B, Elhaty IA, Al HA, Al SR, El-Awady R, Ashraf SS, Amin A. Antioxidant and anticancer activities of Trigonella foenum-graecum, Cassia acutifolia and Rhazya stricta. BMC Complement Altern Med. 2018;18(1):240.PubMedPubMedCentralCrossRef
15.
Al-Dabbagh B, Elhaty IA, Elhaw M, Murali C, Al MA AB, Amin A. Antioxidant and anticancer activities of chamomile (Matricaria recutita L.). BMC Res Notes. 2019;12(1):3.PubMedPubMedCentralCrossRef
16.
El-Dakhly SM, Salama A, Hassanin S, Yassen NN, Hamza AA, Amin A. Aescin and diosmin each alone or in low dose- combination ameliorate liver damage induced by carbon tetrachloride in rats. BMC Res Notes. 2020;13(1):259.PubMedPubMedCentralCrossRef
17.
Yin J, Luo Y, Deng H, Qin S, Tang W, Zeng L, Zhou B. Hugan Qingzhi medication ameliorates hepatic steatosis by activating AMPK and PPARα pathways in L02 cells and HepG2 cells. J Ethnopharmacol. 2014;154(1):229–39.PubMedCrossRef
18.
Tang W, Yao X, Xia F, Yang M, Chen Z, Zhou B, Liu Q. Modulation of the gut microbiota in rats by Hugan Qingzhi tablets during the treatment of high-fat-diet-induced nonalcoholic fatty liver disease. Oxidative Med Cell Longev. 2018;2018:1–14.
19.
Xiao C, He F, Yang M, Xia F, Tang W, Zhou B, Zhou H. Ultra-high performance liquid chromatography/triple quadrupole mass spectrometry study of the stabilities and transformations of four alisols in Alismatis Rhizoma and proprietary traditional Chinese medicine prescriptions. Pharmacogn Mag. 2018;14(58):597–603.CrossRef
20.
Jinjin Y, Waijiao T, Lu Z, Benjie Z. Establishment of a L-02 cell model of hepatic steatosis (in chinese). J Southern Med Univ. 2014;34(06):837–42.
21.
Xiaorui Y, Fan X, Waijiao T, Benjie Z. Effect of Hugan Qingzhi tablets on AMPK pathway activation and NF-κB-p65 protein expression in the liver of rats with nonalcoholic fatty liver disease (in chinese). In. 2017;37:56–62.
22.
Tang W, Zhou B, Zhou H. Study on pharmacodynamics of Hugan Qingzhi tablet on non-alcoholic fatty liver in rats (in chinese). Pharmacol Clin Chin Mater Med. 2013;29(02):169–72.
23.
Tang W, Zeng L, Yin J, Yao Y, Feng L, Yao X, Sun X, Zhou B. Hugan Qingzhi exerts anti-inflammatory effects in a rat model of nonalcoholic fatty liver disease. Evid-Based Complement Alternat Med. 2015;2015:1–13.
24.
Waijiao T, Yufa Y, JInjin Y, Lu Z, Benjie Z. Effects of Hugan Qingzhi tablet on liver protein synthesis and bile metabolism in rats with non-alcoholic fatty liver (in chinese). J Chin Med Mater. 2014;37(08):1452–4.
25.
Xia F, Yao X, Tang W, Xiao C, Yang M, Zhou B. Isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic analysis of Hugan Qingzhi and its protective properties against free fatty acid-induced L02 hepatocyte injury. Front Pharmacol. 2017;8:99.PubMedPubMedCentralCrossRef
26.
Yao X, Xia F, Tang W, Xiao C, Yang M, Zhou B. Isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomics for the investigation of the effect of Hugan Qingzhi on non-alcoholic fatty liver disease in rats. J Ethnopharmacol. 2018;212:208–15.PubMedCrossRef
27.
Perumpail BJ, Khan MA, Yoo ER, Cholankeril G, Kim D, Ahmed A. Clinical epidemiology and disease burden of nonalcoholic fatty liver disease. World J Gastroenterol. 2017;23(47):8263–76.PubMedPubMedCentralCrossRef
28.
29.
Ge J, Wang D, He R, Zhu H, Wang Y, He S. Medicinal herb research: serum pharmacological method and plasma pharmacological method. Biol Pharm Bull. 2010;33(9):1459–65.PubMedCrossRef
30.
Vidyashankar S, Sandeep Varma R, Patki PS. Quercetin ameliorate insulin resistance and up-regulates cellular antioxidants during oleic acid induced hepatic steatosis in HepG2 cells. Toxicol in Vitro. 2013;27(2):945–53.PubMedCrossRef
31.
Soardo G, Donnini D, Domenis L, Catena C, De Silvestri D, Cappello D, Dibenedetto A, Carnelutti A, Bonasia V, Pagano C, et al. Oxidative stress is activated by free fatty acids in cultured human hepatocytes. Metab Syndr Relat Disord. 2011;9(5):397–401.PubMedCrossRef
32.
Gentile CL, Frye M, Pagliassotti MJ. Endoplasmic reticulum stress and the unfolded protein response in nonalcoholic fatty liver disease. Antioxid Redox Sign. 2011;15(2):505–21.
33.
Malhi H, Kaufman RJ. Endoplasmic reticulum stress in liver disease. J Hepatol. 2011;54(4):795–809.PubMedCrossRef
34.
Bobrovnikova-Marjon E, Hatzivassiliou G, Grigoriadou C, Romero M, Cavener DR, Thompson CB, Diehl JA. PERK-dependent regulation of lipogenesis during mouse mammary gland development and adipocyte differentiation. Proc Natl Acad Sci U S A. 2008;105(42):16314–9.PubMedPubMedCentralCrossRef
35.
Kaplowitz N, Than TA, Shinohara M, Ji C. Endoplasmic reticulum stress and liver injury. Semin Liver Dis. 2007;27(4):367–77.PubMedCrossRef
36.
Yeh K, Lai C, Lin C, Hsu C, Lo C, Her GM. ATF4 overexpression induces early onset of hyperlipidaemia and hepatic steatosis and enhances adipogenesis in zebrafsh. Sci Rep. 2017;27(1):16362.CrossRef
37.
Xiao G, Zhang T, Yu S, Lee S, Calabuig-Navarro V, Yamauchi J, Ringquist S, Dong HH. ATF4 protein deficiency protects against high fructose-induced hypertriglyceridemia in mice. J Biol Chem. 2013;288(35):25350–61.PubMedPubMedCentralCrossRef
38.
Wang S, Kaufman RJ. How does protein misfolding in the endoplasmic reticulum affect lipid metabolism in the liver? Curr Opin Lipidol. 2014;25(2):125–32.PubMedCrossRef
39.
So J, Hur KY, Tarrio M, Ruda V, Frank-Kamenetsky M, Fitzgerald K, Koteliansky V, Lichtman AH, Iwawaki T, Glimcher LH, et al. Silencing of lipid metabolism genes through IRE1α-mediated mRNA decay lowers plasma lipids in mice. Cell Metab. 2012;16(4):487–99.PubMedPubMedCentralCrossRef
40.
Sha H, He Y, Chen H, Wang C, Zenno A, Shi H, Yang X, Zhang X, Qi L. The IRE1α-XBP1 pathway of the unfolded protein response is required for Adipogenesis. Cell Metab. 2009;9(6):556–64.PubMedPubMedCentralCrossRef
41.
Maruyama R, Kamoshida Y, Shimizu M, Inoue J, Sato R. ATF6α stimulates Cholesterogenic gene expression and de novo cholesterol synthesis. Biosci Biotechol Biochem. 2013;77(8):1734–8.CrossRef
42.
Mollica MP, Lionetti L, Putti R, Cavaliere G, Gaita M, Barletta A. From chronic overfeeding to hepatic injury_ role of endoplasmic reticulum stress and inflammation. Nutr Metab Cardiovasc Dis. 2011;21:222–30.PubMedCrossRef
43.
Tamaki N, Hatano E, Taura K, Tada M, Kodama Y, Nitta T, Iwaisako K, Seo S, Nakajima A, Ikai I, et al. CHOP deficiency attenuates cholestasis-induced liver fibrosis by reduction of hepatocyte injury. Am J Physiol Gastrointest Liver Physiol. 2008;294(2):G498–505.PubMedCrossRef
44.
Ji C, Mehrian Shai R, Chan C, Hsu YH, Kaplowitz N. Role of CHOP in hepatic apoptosis in the murine model of Intragastric ethanol feeding. Alcohol Clin Exp Res. 2005;29(8):1496–503.PubMedPubMedCentralCrossRef
45.
Yoneda T, Imaizumi K, Oono K, Yui D, Gomi F, Katayama T, Tohyama M. Activation of caspase-12, an endoplastic reticulum (ER) resident caspase, through tumor necrosis factor receptor-associated factor 2-dependent mechanism in response to the ER stress. J Biol Chem. 2001;276(17):13935–40.PubMedCrossRef
46.
Hetz C. Proapoptotic BAX and BAK modulate the unfolded protein response by a direct interaction with IRE1. Science. 2006;312(5773):572–6.PubMedCrossRef
47.
Arruda AP, Pers BM, Parlakgül G, Güney E, Inouye K, Hotamisligil GS. Chronic enrichment of hepatic endoplasmic reticulum-mitochondria contact leads to mitochondrial dysfunction in obesity. Nat Med. 2014;20(12):1427–35.PubMedPubMedCentralCrossRef
48.
Rieusset J. Endoplasmic reticulum-mitochondria calcium signalling in hepatic metabolic diseases. Mol Cell Res. 2017;1864(6):865–76.
49.
Al-Hrout A, Chaiboonchoe A, Khraiwesh B, Murali C, Baig B, El-Awady R, Tarazi H, Alzahmi A, Nelson DR, Greish YE, et al. Safranal induces DNA double-strand breakage and ER-stress-mediated cell death in hepatocellular carcinoma cells. Sci Rep. 2018;8(1):16951.PubMedPubMedCentralCrossRef
50.
Schmitz-Peiffer C, Biden TJ. Protein kinase C function in muscle, liver, and -cells and its therapeutic implications for type 2 Diabetes. Diabetes. 2008;57(7):1774–83.PubMedPubMedCentralCrossRef
51.
Lee SJ, Kang JH, Choi SY, Suk KT, Kim DJ, Kwon O. PKCδ as a regulator for TGFb1-induced a-SMA production in a murine nonalcoholic steatohepatitis model. PLoS One. 2013;8(2):e55979.PubMedPubMedCentralCrossRef
52.
Klymenko K, Novokhatska T, Kizub I, Parshikov A, Dosenko V, Soloviev A. PKC-delta isozyme gene silencing restores vascular function in diabetic rat. J Basic Clin Physiol Pharmacol. 2014;25(4):1–9.CrossRef
53.
Lai S, Li Y, Kuang Y, Cui H, Yang Y, Sun W, Liu K, Chen D, Yan Q, Wen L. PKCδ silencing alleviates saturated fatty acid induced ER stress by enhancing SERCA activity. Biosci Rep. 2017;37(6):R20170869.CrossRef
54.
Luciani DS, Gwiazda KS, Yang TLB, Kalynyak TB, Bychkivska Y, Frey MHZ, Jeffrey KD, Sampaio AV, Underhill TM, Johnson JD. Roles of IP3R and RyR Ca2+ channels in endoplasmic reticulum stress and cell death. Diabetes. 2009;58(2):422–32.PubMedPubMedCentralCrossRef
55.
Wong VK, Li T, Law BY, Ma ED, Yip NC, Michelangeli F, Law CK, Zhang MM, Lam KY, Chan PL, et al. Saikosaponin-d, a novel SERCA inhibitor, induces autophagic cell death in apoptosis-defective cells. Cell Death Dis. 2013;4(7):e720.PubMedPubMedCentralCrossRef
56.
De Ford C, Heidersdorf B, Haun F, Murillo R, Friedrich T, Borner C, Merfort I. The clerodane diterpene casearin J induces apoptosis of T-ALL cells through SERCA inhibition, oxidative stress, and interference with Notch1 signaling. Cell Death Dis. 2016;7(1):e2070.PubMedPubMedCentralCrossRef
57.
Liang C, Han S, Li G, Tabas I, Tall AR. Impaired MEK signaling and SERCA expression promote ER stress and apoptosis in insulin-resistant macrophages and are reversed by Exenatide treatment. Diabetes. 2012;61:2609–20.PubMedPubMedCentralCrossRef
58.
Zhang J, Li Y, Jiang S, Yu H, An W. Enhanced endoplasmic reticulum SERCA activity by overexpression of hepatic stimulator substance gene prevents hepatic cells from ER stress-induced apoptosis. Am J Physiol Cell Physiol. 2014;306(3):C279–90.PubMedCrossRef
59.
Cunha DA, Hekerman P, Ladriere L, Bazarra-Castro A, Ortis F, Wakeham MC, Moore F, Rasschaert J, Cardozo AK, Bellomo E, et al. Initiation and execution of lipotoxic ER stress in pancreatic -cells. J Cell Sci. 2008;121(14):2308–18.PubMedCrossRef
60.
Tong X, Kono T, Evans-Molina C. Nitric oxide stress and activation of AMP-activated protein kinase impair β-cell sarcoendoplasmic reticulum calcium ATPase 2b activity and protein stability. Cell Death Dis. 2015;6(6):e1790.PubMedPubMedCentralCrossRef
Metadata
Title
Hugan Qingzhi medication ameliorates free fatty acid-induced L02 hepatocyte endoplasmic reticulum stress by regulating the activation of PKC-δ
Authors
Miaoting Yang
Zhijuan Chen
Shijian Xiang
Fan Xia
Waijiao Tang
Xiaorui Yao
Benjie Zhou
Publication date
01-12-2020
Publisher
BioMed Central
Published in
BMC Complementary Medicine and Therapies / Issue 1/2020
Electronic ISSN: 2662-7671
DOI
https://doi.org/10.1186/s12906-020-03164-3

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