Nobiletin alleviates palmitic acid‑induced NLRP3 inflammasome activation in a sirtuin 1‑dependent manner in AML‑12 cells

Peng,Zhicheng; Li,Xiaobing; Xing,Dongmei; Du,Xiliang; Wang,Zhe; Liu,Guowen; Li,Xinwei

doi:10.3892/mmr.2018.9615

Nobiletin alleviates palmitic acid‑induced NLRP3 inflammasome activation in a sirtuin 1‑dependent manner in AML‑12 cells

Authors:
- Zhicheng Peng
- Xiaobing Li
- Dongmei Xing
- Xiliang Du
- Zhe Wang
- Guowen Liu
- Xinwei Li
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Affiliations: Key Laboratory of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun, Jilin 130062, P.R. China, Department of Basic Veterinary Medicine, Animal Medicine College, Hunan Agriculture University, Changsha, Hunan 410128, P.R. China
Published online on: October 31, 2018 https://doi.org/10.3892/mmr.2018.9615
Pages: 5815-5822

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Abstract

The NOD‑like receptor family pyrin domain containing 3 (NLRP3) inflammasome has been reported to contribute to palmitic acid (PA)‑induced lipotoxicity. Nobiletin (Nob) is a polymethoxylated flavonoid derived from citrus fruits that has been reported to exert antioxidant and antitumor effects. However, its protective and regulatory mechanisms in PA‑induced lipotoxicity remain unclear. Therefore, the aim of the present study was to investigate the protective effects of Nob in AML‑12 cells against lipotoxicity and examine the underlying mechanism. Western blotting, reverse transcription‑quantitative polymerase chain reaction and ELISA assays were performed to investigate the activation of the NLRP3 inflammasome. Sirtuin 1 (SIRT1) small interfering RNA was used to knockdown SIRT1 expression in AML‑12 cells. The results demonstrated that PA effectively activated NLRP3 inflammasome and increased the expression and secretion of interleukin (IL)‑1β and IL‑18. Notably, the PA‑induced inflammasome activation was reversed by Nob, as indicated by the decreased expression levels of NLRP3, Caspase‑1, IL‑1β and IL‑18. Furthermore, Nob treatment with or without PA enhanced the expression of SIRT1 in AML‑12 cells, while knockdown of SIRT1 with SIRT1‑small interfering RNA reversed the anti‑inflammatory effects of Nob. Overall, the results of the present study indicated that Nob alleviated PA‑induced lipotoxicity in AML‑12 cells via the suppression of NLRP3 inflammasome activation in a SIRT1‑dependent manner. These results provide a possible basis of the underlying mechanism and, in turn, the potential application of Nob in the treatment of non‑alcoholic fatty liver disease.

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1	Ertunc ME and Hotamisligil GS: Lipid signaling and lipotoxicity in metaflammation: Indications for metabolic disease pathogenesis and treatment. J Lipid Res. 57:2099–2114. 2016. View Article : Google Scholar : PubMed/NCBI
2	Du X, Shi Z, Peng Z, Zhao C, Zhang Y, Wang Z and Li X, Liu G and Li X: Acetoacetate induces hepatocytes apoptosis by the ROS-mediated MAPKs pathway in ketotic cows. J Cell Physiol. 232:3296–3308. 2017. View Article : Google Scholar : PubMed/NCBI
3	Nemoto K, Ikeda A, Yoshida C, Kimura J, Mori J, Fujiwara H, Yokosuka A, Mimaki Y, Ohizumi Y and Degawa M: Characteristics of nobiletin-mediated alteration of gene expression in cultured cell lines. Biochem Biophys Res Commun. 431:530–534. 2013. View Article : Google Scholar : PubMed/NCBI
4	Zhang J, Zhang H, Deng X, Zhang Y and Xu K: Baicalin protects AML-12 cells from lipotoxicity via the suppression of ER stress and TXNIP/NLRP3 inflammasome activation. Chem Biol Interact. 278:189–196. 2017. View Article : Google Scholar : PubMed/NCBI
5	Gajaria TK, Patel DK, Devkar RV and Ramachandran AV: Flavonoid rich extract of Murraya Koenigii alleviates in-vitro LDL oxidation and oxidized LDL induced apoptosis in raw 264.7 Murine macrophage cells. J Food Sci Technol. 52:3367–3375. 2015.PubMed/NCBI
6	Kwon EY, Jung UJ, Park T, Yun JW and Choi MS: Luteolin attenuates hepatic steatosis and insulin resistance through the interplay between the liver and adipose tissue in mice with diet-induced obesity. Diabetes. 64:1658–1669. 2015. View Article : Google Scholar : PubMed/NCBI
7	Ye C, Li S, Yao W, Xu L, Qiu Y, Liu Y, Wu Z and Hou Y: The anti-inflammatory effects of baicalin through suppression of NLRP3 inflammasome pathway in LPS-challenged piglet mononuclear phagocytes. Innate Immun. 22:196–204. 2016. View Article : Google Scholar : PubMed/NCBI
8	Vecchione G, Grasselli E, Cioffi F, Baldini F, Oliviera PJ, Sardão VA, Cortese K, Lanni A, Voci A, Portincasa P and Vergani L: The nutraceutic silybin counteracts excess lipid accumulation and ongoing oxidative stress in an in vitro model of non-alcoholic fatty liver disease progression. Front Nutr. 4:422017. View Article : Google Scholar : PubMed/NCBI
9	Loguercio C, Andreone P, Brisc C, Brisc MC, Bugianesi E, Chiaramonte M, Cursaro C, Danila M, de Sio I, Floreani A, et al: Silybin combined with phosphatidylcholine and vitamin E in patients with nonalcoholic fatty liver disease: A randomized controlled trial. Free Radic Biol Med. 52:1658–1665. 2012. View Article : Google Scholar : PubMed/NCBI
10	Wan X, Xu C, Lin Y, Lu C, Li D, Sang J, He H, Liu X, Li Y and Yu C: Uric acid regulates hepatic steatosis and insulin resistance through the NLRP3 inflammasome-dependent mechanism. J Hepatol. 64:925–932. 2016. View Article : Google Scholar : PubMed/NCBI
11	Schroder K, Zhou R and Tschopp J: The NLRP3 inflammasome: A sensor for metabolic danger? Science. 327:296–300. 2010. View Article : Google Scholar : PubMed/NCBI
12	Cai C, Zhu X, Li P, Li J, Gong J, Shen W and He K: NLRP3 deletion inhibits the non-alcoholic steatohepatitis development and inflammation in Kupffer cells induced by palmitic acid. Inflammation. 40:1875–1883. 2017. View Article : Google Scholar : PubMed/NCBI
13	Shirasuna K, Takano H, Seno K, Ohtsu A, Karasawa T, Takahashi M, Ohkuchi A, Suzuki H, Matsubara S, Iwata H and Kuwayama T: Palmitic acid induces interleukin-1β secretion via NLRP3 inflammasomes and inflammatory responses through ROS production in human placental cells. J Reprod Immunol. 116:104–112. 2016. View Article : Google Scholar : PubMed/NCBI
14	Wu J, Xu X, Li Y, Kou J, Huang F, Liu B and Liu K: Quercetin, luteolin and epigallocatechin gallate alleviate TXNIP and NLRP3-mediated inflammation and apoptosis with regulation of AMPK in endothelial cells. Eur J Pharmacol. 745:59–68. 2014. View Article : Google Scholar : PubMed/NCBI
15	Wang W, Wang C, Ding XQ, Pan Y, Gu TT, Wang MX, Liu YL, Wang FM, Wang SJ and Kong LD: Quercetin and allopurinol reduce liver thioredoxin-interacting protein to alleviate inflammation and lipid accumulation in diabetic rats. Br J Pharmacol. 169:1352–1371. 2013. View Article : Google Scholar : PubMed/NCBI
16	Li Y, Yang X, He Y, Wang W, Zhang J, Zhang W, Jing T, Wang B and Lin R: Negative regulation of NLRP3 inflammasome by SIRT1 in vascular endothelial cells. Immunobiology. 222:552–561. 2017. View Article : Google Scholar : PubMed/NCBI
17	Du X, Yang Y, Xu C, Peng Z, Zhang M, Lei L, Gao W, Dong Y, Shi Z, Sun X, et al: Upregulation of miR-181a impairs hepatic glucose and lipid homeostasis. Oncotarget. 8:91362–91378. 2017. View Article : Google Scholar : PubMed/NCBI
18	Fu Y, Wang Y, Du L, Xu C, Cao J, Fan T, Liu J, Su X, Fan S, Liu Q and Fan F: Resveratrol inhibits ionising irradiation-induced inflammation in MSCs by activating SIRT1 and limiting NLRP-3 inflammasome activation. Int J Mol Sci. 14:14105–14118. 2013. View Article : Google Scholar : PubMed/NCBI
19	Li Y, Wang P, Yang X, Wang W, Zhang J, He Y, Zhang W, Jing T, Wang B and Lin R: SIRT1 inhibits inflammatory response partly through regulation of NLRP3 inflammasome in vascular endothelial cells. Mol Immunol. 77:148–156. 2016. View Article : Google Scholar : PubMed/NCBI
20	Kim MY, Lim JH, Youn HH, Hong YA, Yang KS, Park HS, Chung S, Ko SH, Shin SJ, Choi BS, et al: Resveratrol prevents renal lipotoxicity and inhibits mesangial cell glucotoxicity in a manner dependent on the AMPK-SIRT1-PGC1α axis in db/db mice. Diabetologia. 56:204–217. 2013. View Article : Google Scholar : PubMed/NCBI
21	Song Y, Li N, Gu J, Fu S, Peng Z, Zhao C, Zhang Y, Li X, Wang Z, Li X and Liu G: β-Hydroxybutyrate induces bovine hepatocyte apoptosis via an ROS-p38 signaling pathway. J Dairy Sci. 99:9184–9198. 2016. View Article : Google Scholar : PubMed/NCBI
22	Du X, Chen L, Huang D, Peng Z, Zhao C, Zhang Y, Zhu Y, Wang Z, Li X and Liu G: Elevated apoptosis in the liver of dairy cows with ketosis. Cell Physiol Biochem. 43:568–578. 2017. View Article : Google Scholar : PubMed/NCBI
23	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
24	Musso G, Cassader M and Gambino R: Non-alcoholic steatohepatitis: Emerging molecular targets and therapeutic strategies. Nat Rev Drug Discov. 15:249–274. 2016. View Article : Google Scholar : PubMed/NCBI
25	de Alwis NM and Day CP: Non-alcoholic fatty liver disease: The mist gradually clears. J Hepatol. 48 Suppl 1:S104–S112. 2008. View Article : Google Scholar : PubMed/NCBI
26	Hardy T, Anstee QM and Day CP: Nonalcoholic fatty liver disease: New treatments. Curr Opin Gastroenterol. 31:175–183. 2015. View Article : Google Scholar : PubMed/NCBI
27	Michel Anto N, Colberg C, Buscher K, Sommer B, Pramod AB, Ehinger E, Dufner B, Hoppe N, Pfeiffer K, Marchini T, et al: Inflammatory pathways regulated by tumor-necrosis receptor associated factor 1 protect from metabolic consequences in diet-induced obesity. Circ Res. 122:693–700. 2018. View Article : Google Scholar : PubMed/NCBI
28	Vogt BP, Souza PL, Minicucci MF, Martin LC, Barretti P and Caramori JT: Metabolic syndrome criteria as predictors of insulin resistance, inflammation and mortality in chronic hemodialysis patients. Metab Syndr Relat Disord. 12:443–449. 2014. View Article : Google Scholar : PubMed/NCBI
29	Vandanmagsar B, Youm YH, Ravussin A, Galgani JE, Stadler K, Mynatt RL, Ravussin E, Stephens JM and Dixit VD: The NLRP3 inflammasome instigates obesity-induced inflammation and insulin resistance. Nat Med. 17:179–188. 2011. View Article : Google Scholar : PubMed/NCBI
30	Mridha AR, Wree A, Robertson AAB, Yeh MM, Johnson CD, Van Rooyen DM, Haczeyni F, Teoh NC, Savard C, Ioannou GN, et al: NLRP3 inflammasome blockade reduces liver inflammation and fibrosis in experimental NASH in mice. J Hepatol. 66:1037–1046. 2017. View Article : Google Scholar : PubMed/NCBI
31	Van De Wier B, Koek GH, Bast A and Haenen GR: The potential of flavonoids in the treatment of non-alcoholic fatty liver disease. Crit Rev Food Sci Nutr. 57:834–855. 2017. View Article : Google Scholar : PubMed/NCBI
32	Sun X, Yuan X, Chen L, Wang T, Wang Z, Sun G, Li X, Li X and Liu G: Histamine induces bovine rumen epithelial cell inflammatory response via NF-κB pathway. Cell Physiol Biochem. 42:1109–1119. 2017. View Article : Google Scholar : PubMed/NCBI
33	Zhang N, Yang Z, Xiang SZ, Jin YG, Wei WY, Bian ZY, Deng W and Tang QZ: Nobiletin attenuates cardiac dysfunction, oxidative stress, and inflammatory in streptozotocin: Induced diabetic cardiomyopathy. Mol Cell Biochem. 417:87–96. 2016. View Article : Google Scholar : PubMed/NCBI
34	He Z, Li X, Chen H, He K, Liu Y and Gong J and Gong J: Nobiletin attenuates lipopolysaccharide/D-galactosamineinduced liver injury in mice by activating the Nrf2 antioxidant pathway and subsequently inhibiting NF-κB-mediated cytokine production. Mol Med Rep. 14:5595–5600. 2016. View Article : Google Scholar : PubMed/NCBI
35	Purushotham A, Schug TT, Xu Q, Surapureddi S, Guo X and Li X: Hepatocyte-specific deletion of SIRT1 alters fatty acid metabolism and results in hepatic steatosis and inflammation. Cell Metab. 9:327–338. 2009. View Article : Google Scholar : PubMed/NCBI
36	Li Y, Xu S, Giles A, Nakamura K, Lee JW, Hou X, Donmez G, Li J, Luo Z, Walsh K, et al: Hepatic overexpression of SIRT1 in mice attenuates endoplasmic reticulum stress and insulin resistance in the liver. FASEB J. 25:1664–1679. 2011. View Article : Google Scholar : PubMed/NCBI