Artesunate exerts protective effects against ulcerative colitis via suppressing Toll‑like receptor 4 and its downstream nuclear factor‑κB signaling pathways

Chen,Yu‑Xuan; Zhang,Xiao‑Qi; Yu,Cheng‑Gong; Huang,Shu‑Ling; Xie,Ying; Dou,Xiao‑Tan; Liu,Wen‑Jia; Zou,Xiao‑Ping

doi:10.3892/mmr.2019.10345

Artesunate exerts protective effects against ulcerative colitis via suppressing Toll‑like receptor 4 and its downstream nuclear factor‑κB signaling pathways

Authors:
- Yu‑Xuan Chen
- Xiao‑Qi Zhang
- Cheng‑Gong Yu
- Shu‑Ling Huang
- Ying Xie
- Xiao‑Tan Dou
- Wen‑Jia Liu
- Xiao‑Ping Zou
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Affiliations: Department of Gastroenterology, Nanjing Drum Tower Hospital, Nanjing University, Nanjing, Jiangsu 210000, P.R. China
Published online on: June 5, 2019 https://doi.org/10.3892/mmr.2019.10345
Pages: 1321-1332
Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Artesunate (ART) is a semi‑synthetic derivative of artemisinin used in the treatment of patients with malaria, which has also been reported to have immunoregulatory, anticancer and anti‑inflammatory properties. The aim of the present study was to investigate the possible beneficial effects of ART on ulcerative colitis (UC) rats and to detect the possible mechanisms underlying these effects. A UC rat model was established using dextran sulfate sodium (DSS). Rats were randomly divided into the following groups: Normal control, UC model group, UC rats treated with a low, medium or high dose of ART (10, 30 and 50 mg/kg/day, respectively), and the positive control group (50 mg/kg/day 5‑aminosalicylic acid). The damage status of colonic mucosal epithelial tissue was investigated by hematoxylin and eosin staining, and then the weight, colon length and disease activity index (DAI) were measured. Western blotting and reverse transcription‑quantitative polymerase chain reaction analysis were used to detect the levels of cytokines associated with UC and proteins associated with Toll‑like receptor 4 (TLR4)‑nuclear factor (NF)‑κB pathway. ELISA was also performed to measure the levels of inflammatory cytokines. In addition, the viability and infiltration of RAW264.7 cells were examined using Cell Counting Kit‑8 and Transwell assays. The results demonstrated that treatment with ART significantly alleviated the UC symptoms induced by DSS in the rat model, lowered the DAI, ameliorated pathological changes, attenuated colon shortening, inhibited the levels of pro‑inflammatory mediators and myeloperoxidase activity, and increased hemoglobin expression. Additionally, inflammatory and apoptotic markers were found to be significantly downregulated following treatment with ART in UC rats and RAW264.7 cells. To the best of our knowledge, the present study is the first to demonstrate that ART exerts anti‑inflammatory effects via regulating the TLR4‑NF‑κB signaling pathway in UC.

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1	Ito A, Omori T, Hanafusa N, Tsuchiya K, Nakamura S and Tokushige K: Efficacy and safety of granulocyte adsorption apheresis in elderly patients with ulcerative colitis. J Clin Apher. 33:514–520. 2018. View Article : Google Scholar : PubMed/NCBI
2	Høivik ML, Moum B, Solberg IC, Henriksen M, Cvancarova M and Bernklev T; IBSEN Group, : Work disability in inflammatory bowel disease patients 10 years after disease onset: Results from the IBSEN Study. Gut. 62:368–375. 2013. View Article : Google Scholar : PubMed/NCBI
3	Torres J, Billioud V, Sachar DB, Peyrin-Biroulet L and Colombel JF: Ulcerative colitis as a progressive disease: The forgotten evidence. Inflamm Bowel Dis. 18:1356–1363. 2012. View Article : Google Scholar : PubMed/NCBI
4	Ungaro R, Mehandru S, Allen PB, Peyrin-Biroulet L and Colombel JF: Ulcerative colitis. Lancet. 389:1756–1770. 2017. View Article : Google Scholar : PubMed/NCBI
5	Tsuda K, Miyamoto L, Hamano S, Morimoto Y, Kangawa Y, Fukue C, Kagawa Y, Horinouchi Y, Xu W, Ikeda Y, et al: Mechanisms of the pH- and oxygen-dependent oxidation activities of artesunate. Biol Pharm Bull. 41:555–563. 2018. View Article : Google Scholar : PubMed/NCBI
6	Efferth T, Giaisi M, Merling A, Krammer PH and Li-Weber M: Artesunate induces ROS-mediated apoptosis in doxorubicin-resistant T leukemia cells. PLoS One. 2:e6932007. View Article : Google Scholar : PubMed/NCBI
7	Ooko E, Saeed ME, Kadioglu O, Sarvi S, Colak M, Elmasaoudi K, Janah R, Greten HJ and Efferth T: Artemisinin derivatives induce iron-dependent cell death (ferroptosis) in tumor cells. Phytomedicine. 22:1045–1054. 2015. View Article : Google Scholar : PubMed/NCBI
8	Button RW, Lin F, Ercolano E, Vincent JH, Hu B, Hanemann CO and Luo S: Artesunate induces necrotic cell death in schwannoma cells. Cell Death Dis. 5:e14662014. View Article : Google Scholar : PubMed/NCBI
9	Berdelle N, Nikolova T, Quiros S, Efferth T and Kaina B: Artesunate induces oxidative DNA damage, sustained DNA double-strand breaks, and the ATM/ATR damage response in cancer cells. Mol Cancer Ther. 10:2224–2233. 2011. View Article : Google Scholar : PubMed/NCBI
10	Eling N, Reuter L, Hazin J, Hamacher-Brady A and Brady NR: Identification of artesunate as a specific activator of ferroptosis in pancreatic cancer cells. Oncoscience. 2:517–532. 2015. View Article : Google Scholar : PubMed/NCBI
11	Cen Y, Liu C, Li X, Yan Z, Kuang M, Su Y, Pan X, Qin R, Liu X, Zheng J and Zhou H: Artesunate ameliorates severe acute pancreatitis (SAP) in rats by inhibiting expression of pro-inflammatory cytokines and Toll-like receptor 4. Int Immunopharmacol. 38:252–260. 2016. View Article : Google Scholar : PubMed/NCBI
12	Sands BE and Kaplan GG: The role of TNFalpha in ulcerative colitis. J Clin Pharmacol. 47:930–941. 2007. View Article : Google Scholar : PubMed/NCBI
13	Allocca M, Furfaro F, Fiorino G, Gilardi D, D'Alessio S and Danese S: Can IL-23 be a good target for ulcerative colitis? Best Pract Res Clin Gastroenterol. 32-33:95–102. 2018. View Article : Google Scholar : PubMed/NCBI
14	Jeoung BR, Lee KD, Na CS, Kim YE, Kim B and Kim YR: Ganghwaljetongyeum, an anti-arthritic remedy, attenuates synoviocyte proliferation and reduces the production of proinflammatory mediators in macrophages: The therapeutic effect of GHJTY on rheumatoid arthritis. BMC Complement Altern Med. 13:472013. View Article : Google Scholar : PubMed/NCBI
15	Jeong HY, Choi YS, Lee JK, Lee BJ, Kim WK and Kang H: Anti-inflammatory activity of citric acid-treated wheat germ extract in lipopolysaccharide-stimulated macrophages. Nutrients. 9:2017. View Article : Google Scholar
16	Xiang Y, Ye W, Huang C, Lou B, Zhang J, Yu D, Huang X, Chen B and Zhou M: Brusatol inhibits growth and induces apoptosis in pancreatic cancer cells via JNK/p38 MAPK/NF-κb/Stat3/Bcl-2 signaling pathway. Biochem Biophys Res Commun. 487:820–826. 2017. View Article : Google Scholar : PubMed/NCBI
17	Xie Z, Xiao Z and Wang F: Hepatitis C virus nonstructural 5A protein (HCV-NS5A) inhibits hepatocyte apoptosis through the NF-κb/miR-503/bcl-2 pathway. Mol Cells. 40:202–210. 2017.PubMed/NCBI
18	Eissa N, Hussein H, Kermarrec L, Elgazzar O, Metz-Boutigue MH, Bernstein CN and Ghia JE: Chromofungin (CHR: CHGA_47-66) is downregulated in persons with active ulcerative colitis and suppresses pro-inflammatory macrophage function through the inhibition of NF-κB signaling. Biochem Pharmacol. 145:102–113. 2017. View Article : Google Scholar : PubMed/NCBI
19	Gu P, Zhu L, Liu Y, Zhang L, Liu J and Shen H: Protective effects of paeoniflorin on TNBS-induced ulcerative colitis through inhibiting NF-kappaB pathway and apoptosis in mice. Int Immunopharmacol. 50:152–160. 2017. View Article : Google Scholar : PubMed/NCBI
20	Ding X, Liang Y, Peng W, Li R, Lin H, Zhang Y and Lu D: Intracellular TLR22 acts as an inflammation equalizer via suppression of NF-κB and selective activation of MAPK pathway in fish. Fish Shellfish Immunol. 72:646–657. 2018. View Article : Google Scholar : PubMed/NCBI
21	Pan T, Shi X, Chen H, Chen R, Wu D, Lin Z, Zhang J and Pan J: Geniposide suppresses interleukin-1β-induced inflammation and apoptosis in rat chondrocytes via the PI3K/Akt/NF-κB signaling pathway. Inflammation. 41:390–399. 2018. View Article : Google Scholar : PubMed/NCBI
22	Liu B, Li S, Sui X, Guo L, Liu X, Li H, Gao L, Cai S, Li Y, Wang T and Piao X: Root extract of polygonum cuspidatum Siebold & Zucc. ameliorates DSS-induced ulcerative colitis by affecting NF-kappaB signaling pathway in a mouse model via synergistic effects of polydatin, resveratrol, and emodin. Front Pharmacol. 9:3472018. View Article : Google Scholar : PubMed/NCBI
23	Manolakis AC, Kapsoritakis AN, Kapsoritaki A, Tiaka EK, Oikonomou KA, Lotis V, Vamvakopoulou D, Davidi I, Vamvakopoulos N and Potamianos SP: Readressing the role of toll-like receptor-4 alleles in inflammatory bowel disease: Colitis, smoking, and seroreactivity. Dig Dis Sci. 58:371–380. 2013.PubMed/NCBI
24	Liu Y, Zhang Z, Wang L, Li J, Dong L, Yue W, Chen J, Sun X, Zhong L and Sun D: TLR4 monoclonal antibody blockade suppresses dextran-sulfate-sodium-induced colitis in mice. J Gastroenterol Hepatol. 25:209–214. 2010. View Article : Google Scholar : PubMed/NCBI
25	Castelli AA, Estrada JJ and Kaminski JP: Patient with ulcerative colitis and abdominal pain. JAMA Surg. 153:282–283. 2018. View Article : Google Scholar : PubMed/NCBI
26	Murthy SN, Cooper HS, Shim H, Shah RS, Ibrahim SA and Sedergran DJ: Treatment of dextran sulfate sodium-induced murine colitis by intracolonic cyclosporin. Dig Dis Sci. 38:1722–1734. 1993. View Article : Google Scholar : PubMed/NCBI
27	Cooper HS, Murthy SN, Shah RS and Sedergran DJ: Clinicopathologic study of dextran sulfate sodium experimental murine colitis. Lab Invest. 69:238–249. 1993.PubMed/NCBI
28	Yang YH, Li DL, Bi XY, Sun L, Yu XJ, Fang HL, Miao Y, Zhao M, He X, Liu JJ and Zang WJ: Acetylcholine inhibits LPS-induced MMP-9 production and cell migration via the α7 nAChR-JAK2/STAT3 pathway in RAW264.7 cells. Cell Physiol Biochem. 36:2025–2038. 2015. View Article : Google Scholar : PubMed/NCBI
29	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
30	Zhong W, Lu X, Shi H, Zhao G, Song Y, Wang Y, Zhang J, Jin Y and Wang S: Distinct microbial populations exist in the mucosa-associated microbiota of diarrhea predominant irritable bowel syndrome and ulcerative colitis. J Clin Gastroenterol. 28–Nov;2017.doi: 10.1097/MCG.0000000000000961 (Epub ahead of print). View Article : Google Scholar
31	Nigg, Kolyvanos N, Käser and Vetter: Colitis ulcerosa. Praxis. 97:167–174. 2008. View Article : Google Scholar : PubMed/NCBI
32	Shimodate Y, Takanashi K, Waga E, Fujita T, Katsuki S and Nomura M: Exacerbation of bloody diarrhea as a side effect of mesalamine treatment of active ulcerative colitis. Case Rep Gastroenterol. 5:159–165. 2011. View Article : Google Scholar : PubMed/NCBI
33	Muller AF, Stevens PE, McIntyre AS, Ellison H and Logan RF: Experience of 5-aminosalicylate nephrotoxicity in the United Kingdom. Aliment Pharmacol Ther. 21:1217–1224. 2005. View Article : Google Scholar : PubMed/NCBI
34	Ericsson T, Blank A, von Hagens C, Ashton M and Äbelö A: Population pharmacokinetics of artesunate and dihydroartemisinin during long-term oral administration of artesunate to patients with metastatic breast cancer. Eur J Clin Pharmacol. 70:1453–1463. 2014. View Article : Google Scholar : PubMed/NCBI
35	Anfosso L, Efferth T, Albini A and Pfeffer U: Microarray expression profiles of angiogenesis-related genes predict tumor cell response to artemisinins. Pharmacogenomics J. 6:269–278. 2006. View Article : Google Scholar : PubMed/NCBI
36	Efferth T, Sauerbrey A, Olbrich A, Gebhart E, Rauch P, Weber HO, Hengstler JG, Halatsch ME, Volm M, Tew KD, et al: Molecular modes of action of artesunate in tumor cell lines. Mol Pharmacol. 64:382–394. 2003. View Article : Google Scholar : PubMed/NCBI
37	Efferth T, Ramirez T, Gebhart E and Halatsch ME: Combination treatment of glioblastoma multiforme cell lines with the anti-malarial artesunate and the epidermal growth factor receptor tyrosine kinase inhibitor OSI-774. Biochem Pharmacol. 67:1689–1700. 2004. View Article : Google Scholar : PubMed/NCBI
38	Kumar H, Kawai T and Akira S: Toll-like receptors and innate immunity. Biochem Biophys Res Commun. 388:621–625. 2009. View Article : Google Scholar : PubMed/NCBI
39	Chi W, Chen H, Li F, Zhu Y, Yin W and Zhuo Y: HMGB1 promotes the activation of NLRP3 and caspase-8 inflammasomes via NF-κB pathway in acute glaucoma. J Neuroinflammation. 12:1372015. View Article : Google Scholar : PubMed/NCBI
40	Saluk J, Bijak M, Posmyk M and Zbikowska H: Red cabbage anthocyanins as inhibitors of lipopolysaccharide-induced oxidative stress in blood platelets. Int J Biol Macromol. 80:702–709. 2015. View Article : Google Scholar : PubMed/NCBI
41	Martinou JC and Green DR: Breaking the mitochondrial barrier. Nat Rev Mol Cell Biol. 2:63–67. 2001. View Article : Google Scholar : PubMed/NCBI
42	Siddiqui WA, Ahad A and Ahsan H: The mystery of BCL2 family: Bcl-2 proteins and apoptosis: An update. Arch Toxicol. 89:289–317. 2015. View Article : Google Scholar : PubMed/NCBI
43	Nontprasert A, Pukrittayakamee S, Dondorp AM, Clemens R, Looareesuwan S and White NJ: Neuropathologic toxicity of artemisinin derivatives in a mouse model. Am J Trop Med Hyg. 67:423–429. 2002. View Article : Google Scholar : PubMed/NCBI