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Journal of Integrative Medicine ›› 2016, Vol. 14 ›› Issue (3): 203-208.doi: 10.1016/S2095-4964(16)60257-1

• Research Article • Previous Articles     Next Articles

Celastrol targets IRAKs to block Toll-like receptor 4-mediated nuclear factor-κB activation

Yu-fanShena,b, XueZhangb, YingWangb, Fan-fanCaob, GeorgesUzanb,c, BinPengd , Deng-haiZhangb,c   

  1. a Department of Clinical Laboratory Diagnostics, Postgraduate Education College, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
    b Sino-French Cooperative Central Laboratory, Shanghai Gongli Hospital, Second Military Medical University, Shanghai 200135, China
    c U972, Inserm, Lavoisier Building, Paul Brousse Hospital, 94807, Villejuif Cedex, France
  • Received:2016-01-07 Accepted:2016-03-08 Online:2016-05-15 Published:2016-05-15
  • Contact: Bin Peng; E-mail: Deng-hai Zhang, Professor, PhD, MD; E-mail:


Celastrol has been established as a nuclear factor-κB (NF-κB) activation inhibitor; however, the exact mechanism behind this action is still unknown. Using text-mining technology, the authors predicted that interleukin-1 receptor-associated kinases (IRAKs) are potential celastrol targets, and hypothesized that targeting IRAKs might be one way that celastrol inhibits NF-κB. This is because IRAKs are key molecules for some crucial pathways to activate NF-κB (e.g., the interleukin-1 receptor (IL-1R)/Toll-like receptor (TLR) superfamily).


The human hepatocellular cell line (HepG2) treated with palmitic acid (PA) was used as a model for stimulating TLR4/NF-κB activation, in order to observe the potential effects of celastrol in IRAK regulation and NF-κB inhibition. The transfection of small interfering RNA was used for down-regulating TLR4, IRAK1 and IRAK4, and the Western blot method was used to detect changes in the protein expressions.


The results showed that celastrol could effectively inhibit PA-caused TLR4-dependent NF-κB activation in the HepG2 cells; PA also activated IRAKs, which were inhibited by celastrol. Knocking down IRAKs abolished PA-caused NF-κB activation.


The results for the first time show that targeting IRAKs is one way in which celastrol inhibits NF-κB activation.

Key words: Celastrol, Interleukin-1 receptor-associated kinases, Nuclear factor-kappa B, Toll-like receptor 4, Hepatocytes

1 Zhang DH, Marconi A, Xu LM, Yang CX, Sun GW, Feng XL, Ling CQ, Qin WZ, Uzan G, d’Alessio P. Tripterine inhibits the expression of adhesion molecules in activated endothelial cells. [J] Leukoc Biol. 2006; 80(2): 309-319.
2 Li HY, Zhang J, Sun LL, Li BH, Gao HL, Xie T, Zhang N, Ye ZM.Celastrol induces apoptosis and autophagy via the ROS/ JNK signaling pathway in human osteosarcoma cells: an in vitro and in vivo study. Cell Death Dis. 2015; 6: e1604.
3 Konieczny J, Jantas D, Lenda T, Domin H, Czarnecka A, Kuter K, Smialowska M, Lason W, Lorenc-Koci E.Lack of neuroprotective effect of celastrol under conditions of proteasome inhibition by lactacystin in vitro and in vivo studies: implications for Parkinson’s disease. Neurotox Res. 2014; 26(3): 255-273.
4 Liu J, Lee J, Salazar Hernandez MA, Mazitschek R, Ozcan U.Treatment of obesity with celastrol. Cell. 2015; 161(5): 999-1011.
5 Kim JE, Lee MH, Nam DH, Song HK, Kang YS, Lee JE, Kim HW, Cha JJ, Hyun YY, Han SY, Han KH, Han JY, Cha DR.Celastrol, an NF-κB inhibitor, improves insulin resistance and attenuates renal injury in db/db mice. PLoS One. 2013; 8(4): e62068.
6 Lu C, Zhang X, Zhang D, Pei E, Xu J, Tang T, Ye M, Uzan G, Zhi K, Li M, Zuo K.Short time tripterine treatment enhances endothelial progenitor cell function via heat shock protein 32. [J] Cell Physiol. 2015; 230(5): 1139-1147.
7 Li-Weber M.Targeting apoptosis pathways in cancer by Chinese medicine. Cancer Lett. 2013; 332(2): 304-312.
8 Nam NH.Naturally occurring NF-kB inhibitors. Mini Rev Med Chem. 2006; 6(8): 945-951.
9 Jin HZ, Hwang BY, Kim HS, Lee JH, Kim YH, Lee JJ.Antiinflammatory constituents of Celastrus orbiculatus inhibit the NF-kB activation and NO production. [J] Nat Prod. 2002; 65(1): 89-91.
10 Lee JH, Koo TH, Yoon H, Jung HS, Jin HZ, Lee K, Hong YS, Lee JJ.Inhibition of NF-kB activation through targeting IkB kinase by celastrol, a quinone methide triterpenoid. Biochem Pharmacol. 2006; 72(10): 1311-1321.
11 Sethi G, Ahn KS, Pandey MK, Aggarwal BB.Celastrol, a novel triterpene, potentiates TNF-induced apoptosis and suppresses invasion of tumor cells by inhibiting NF-kB-regulated gene products and TAK1-mediated NF-kB activation. Blood. 2007; 109(7): 2727-2735.
12 Torvik VI, Smalheiser NR.A quantitative model for linking two disparate sets of articles in MEDLINE. Bioinformatics. 2007; 23(13): 1658-1665.
13 Smalheiser NR, Torvik VI, Zhou W.Arrowsmith two-node search interface: a tutorial on finding meaningful links between two disparate sets of articles in MEDLINE. Comput Methods Programs Biomed. 2009; 94(2): 190-197.
14 Martin MU, Wesche H.Summary and comparison of the signaling mechanisms of the Toll/interleukin-1 receptor family. Biochim Biophys Acta. 2002; 1592(3): 265-280.
15 Dziarski R, Gupta D.Role of MD-2 in TLR2- and TLR4- mediated recognition of Gram-negative and Gram-positive bacteria and activation of chemokine genes. [J] Endotoxin Res. 2000; 6(5): 401-405.
16 Jin J, Zhang X, Lu Z, Perry DM, Li Y, Russo SB, Cowart LA, Hannun YA, Huang Y.Acid sphingomyelinase plays a key role in palmitic acid-amplified inflammatory signaling triggered by lipopolysaccharide at low concentrations in macrophages. Am [J] Physiol Endocrinol Metab. 2013; 305(7): E853-E867.
17 Schaeffler A, Gross P, Buettner R, Bollheimer C, Buechler C, Neumeier M, Kopp A, Schoelmerich J, Falk W.Fatty acid-induced induction of Toll-like receptor-4/nuclear factor-kB pathway in adipocytes links nutritional signalling with innate immunity. Immunology. 2009; 126(2): 233-245.
18 Maloney E, Sweet IR, Hockenbery DM, Pham M, Rizzo NO, Tateya S, Handa P, Schwartz MW, Kim F.Activation of NF-kB by palmitate in endothelial cells: a key role for NADPH oxidase-derived superoxide in response to TLR4 activation. Arterioscler Thromb Vasc Biol. 2009; 29(9): 1370-1375.
19 Sharifnia T, Antoun J, Verriere TG, Suarez G, Wattacheril J, Wilson KT, Peek RM, Jr., Abumrad NN, Flynn CR. Hepatic TLR4 signaling in obese NAFLD. Am [J] Physiol Gastrointest Liver Physiol. 2015; 309(4): G270-G278.
20 Cousin SP, Hugl SR, Wrede CE, Kajio H, Myers MG Jr., Rhodes CJ.Free fatty acid-induced inhibition of glucose and insulin-like growth factor I-induced deoxyribonucleic acid synthesis in the pancreatic b-cell line INS-1. Endocrinology. 2001; 142(1): 229-240.
21 Suzuki N, Suzuki S, Duncan GS, Millar DG, Wada T, Mirtsos C, Takada H, Wakeham A, Itie A, Li S, Penninger JM, Wesche H, Ohashi PS, Mak TW, Yeh WC.Severe impairment of interleukin-1 and Toll-like receptor signalling in mice lacking IRAK-4. Nature. 2002; 416(6882): 750-756.
22 Rhyasen GW, Starczynowski DT.IRAK signalling in cancer. Br [J] Cancer. 2015; 112(2): 232-237.
23 Lin SC, Lo YC, Wu H.Helical assembly in the MyD88- IRAK4-IRAK2 complex in TLR/IL-1R signalling. Nature. 2010; 465(7300): 885-890.
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