Screening of Bufadienolides from Toad Venom Identifies Gammabufotalin as a Potential Anti-inflammatory Agent

Abstract

Toad venom (Chansu) is used in the treatment of infectious and inflammatory diseases in China and East/Southeast Asian countries. However, the anti-inflammatory components of toad venom have not yet been systematically evaluated and clearly defined. To investigate the anti-inflammatory effects of toad venom and identify new anti-inflammatory ingredients, we used zebrafish, an alternative drug screening model, to evaluate the anti-inflammatory effects of 14 bufadienolides previously isolated from toad venom. Most of the bufadienolides were found to exert significant anti-inflammatory effects on lipopolysaccharide-, CuSO₄-, or tail transection-induced zebrafish inflammatory models. Moreover, gammabufotalin (

6) inhibits lipopolysaccharide-induced inflammation by suppressing the myeloid differentiation primary response 88/nuclear factor-kappa B and STAT3 signal pathways. This study confirms the potential of zebrafish in drug screening, clarifies the anti-inflammatory effects of bufadienolides from toad venom, and indicates that gammabufotalin may be developed as a novel therapeutic agent for inflammatory diseases in the future.

Key words

bufadienolides - gammabufotalin - bufonidae - Bufo gargarizans - anti-inflammation - zebrafish - Danio rerio

Publikationsverlauf

Eingereicht: 01. Februar 2020

Angenommen nach Revision: 23. August 2020

Artikel online veröffentlicht:
13. Oktober 2020

• References

• 1 Zhang DM, Liu JS, Deng LJ, Chen MF, Yiu A, Cao HH, Tian HY, Fung KP, Kurihara H, Pan JX, Ye WC. Arenobufagin, a natural bufadienolide from toad venom, induces apoptosis and autophagy in human hepatocellular carcinoma cells through inhibition of PI3K/Akt/mTOR pathway. Carcinogenesis 2013; 34: 1331-1342
• 2 Liu JS, Zhang DM, Li Y, Chen WM, Ruan ZX, Deng LJ, Wang LW, Tian HY, Yiu A, Fan CL, Luo H, Liu SW, Wang Y, Xiao GK, Chen LX, Ye WC. Discovery of bufadienolides as a novel class of ClC-3 chloride channel activators with antitumor activities. J Med Chem 2013; 56: 5734-5743
• 3 Chen KK, Kovarikova A. Pharmacology and toxicology of toad venom. J Pharm Sci 1967; 56: 1535-1541
• 4 Qi J, Tan CK, Hashimi SM, Zulfiker AH, Good D, Wei MQ. Toad glandular secretions and skin extractions as anti-inflammatory and anticancer agents. Evid Based Complement Alternat Med 2014; 2014: 312684
• 5 Chinese Pharmacopoeia Commission. Chinese Pharmacopoeia, 10th ed. Beijing: China Medical Science and Technology Press; 2015: 383-384
• 6 Wen LL, Huang Y, Xie XB, Huang W, Yin JQ, Lin WQ, Jia Q, Zeng WA. Anti-inflammatory and antinociceptive activities of bufalin in rodents. Mediators Inflamm 2014; 2014: 171839
• 7 Chen YY, Lu HF, Hsu SC, Kuo CL, Chang SJ, Lin JJ, Wu PP, Liu JY, Lee CH, Chung JG, Chang JB. Bufalin inhibits migration and invasion in human hepatocellular carcinoma SK-Hep1 cells through the inhibitions of NF-kB and matrix metalloproteinase-2/-9-signaling pathways. Environ Toxicol 2015; 30: 74-82
• 8 Xie SS, Spelmink L, Codemo M, Subramanian K, Pütsep K, Henriques-Normark B, Olliver M. Cinobufagin modulates human innate immune responses and triggers antibacterial activity. PLoS One 2016; 11: e0160734
• 9 MacRae CA, Peterson RT. Zebrafish as tools for drug discovery. Nat Rev Drug Discov 2015; 14: 721-731
• 10 Ganzen L, Venkatraman P, Pang CP, Leung YF, Zhang MZ. Utilizing zebrafish visual behaviors in drug screening for retinal degeneration. Int J Mol Sci 2017; 18: 1185
• 11 Chua SC, Tan HQ, Engelberg D, Engelberg D, Lim L. Alternative experimental models for studying influenza proteins, host-virus interactions and anti-influenza drugs. Pharm 2019; 12: 147
• 12 Trede NS, Langenau DM, Traver D, Look AT, Zon LI. The use of zebrafish to understand immunity. Immunity 2004; 20: 367-379
• 13 Rojo I, de Ilárduya OM, Estonba A, Pardo MA. Innate immune gene expression in individual zebrafish after Listonella anguillarum inoculation. Fish Shellfish Immunol 2007; 23: 1285-1293
• 14 Correa RG, Tergaonkar V, Ng JK, Dubova I, Izpisua-Belmonte JC, Verma IM. Characterization of NF-kappa B/I kappa B proteins in zebra fish and their involvement in notochord development. Mol Cell Biol 2004; 24: 5257-5268
• 15 Rahman A, Fazal F. Blocking NF-κB: an inflammatory issue. Proc Am Thorac Soc 2011; 8: 497-503
• 16 Guo D, Li JR, Wang Y, Lei LS, Yu CL, Chen NN. Cyclovirobuxinum D suppresses lipopolysaccharide-induced inflammatory responses in murine macrophages in vitro by blocking JAK-STAT signaling pathway. Acta Pharmacol Sin 2014; 35: 770-778
• 17 Liu JS, Deng LJ, Tian HY, Ruan ZX, Cao HH, Ye WC, Zhang DM, Yu ZL. Anti-tumor effects and 3D-quantitative structure-activity relationship analysis of bufadienolides from toad venom. Fitoterapia 2019; 134: 362-371
• 18 Renshaw S, Loynes C, Trushell D, Elworthy S, Ingham P, Whyte M. A transgenic zebrafish model of neutrophilic inflammation. Blood 2006; 108: 3976-3979
• 19 Yang LL, Wang GQ, Yang LM, Huang ZB, Zhang WQ, Yu LZ. Endotoxin molecule lipopolysaccharide-induced zebrafish inflammation model: a novel screening method for anti-inflammatory drugs. Molecules 2014; 19: 2390-2409
• 20 Gan L, Zheng YR, Deng LJ, Sun PH, Ye JX, Wei XD, Liu FF, Yu LZ, Ye WC, Fan CL, Liu JS, Zhang WQ. Diterpenoid lactones with anti-inflammatory effects from the aerial parts of Andrographis paniculate . Molecules 2019; 24: 2726
• 21 Yang LL, Zhou XJ, Huang WJ, Fang Q, Hu JL, Yu LZ, Ma N, Zhang WQ. Protective effect of phillyrin on lethal LPS-induced neutrophil inflammation in zebrafish. Cell Physiol Biochem 2017; 43: 2074-2087
• 22 Kim GR, Yang JY, Hwang KS, Kim SS, Chae JS, Kan H, Ahn JH, Lee WM, Ahn SH, Lee YM, Bae MA, Shin DS. Anti-inflammatory effect of a novel synthetic compound 1-((4-fluorophenyl)thio) isoquinoline in RAW264.7 macrophages and a zebrafish model. Fish Shellfish Immunol 2019; 87: 395-400
• 23 DʼAlenon CA, Pea OA, Wittmann C, Gallardo VE, Jones RA, Loosli F, Liebel U, Grabher C, Allende ML. A high-throughput chemically induced inflammation assay in zebrafish. BMC Biol 2010; 8: 151
• 24 Li LL, Zhang S, Xin YF, Sun JY, Xie F, Yang L, Chen ZQ, Chen H, Liu F, Xuan YX. Role of Quzhou fructus Aurantii extract in preventing and treating acute lung injury and inflammation. Sci Rep 2018; 8: 1698
• 25 Huang MY, Lin J, Lu K, Xu HG, Geng ZZ, Sun PH, Chen WM. Anti-inflammatory effects of cajaninstilbene acid and its derivatives. J Agric Food Chem 2016; 64: 2893-2900
• 26 Fan HY, Gao ZF, Ji K, Li X, Wu JB, Liu Y, Wang XK, Liang HY, Liu YN, Li XT, Liu P, Chen DQ, Zhao F. The in vitro and in vivo anti-inflammatory effect of osthole, the major natural coumarin from Cnidium monnieri (L.) Cuss, via the blocking of the activation of the NF-κB and MAPK/p38 pathways. Phytomedicine 2019; 18: 1-11
• 27 Varela M, Dios S, Novoa B, Figueras A. Characterisation, expression and ontogeny of interleukin-6 and its receptors in zebrafish (Danio rerio). Dev Comp Immunol 2012; 37: 97-106
• 28 Kallen KJ. The role of transsignalling via the agonistic soluble IL-6 receptor in human diseases. Biochim Biophys Acta 2002; 1592: 323-343
• 29 Cheng BC, Ma XQ, Kwan HY, Tse KW, Cao HH, Su T, Shu X, Wu ZZ, Yu ZL. An herbal formula consisting of Rosae Multiflorae Fructus and Lonicerae Japonicae Flos inhibits inflammatory mediators in LPS-stimulated RAW 264.7 macrophages. J Ethnopharmacol 2014; 153: 922-927
• 30 OʼNeill LA, Bowie AG. The family of five: TIR-domain-containing adaptors in Toll-like receptor signaling. Nat Rev Immunol 2007; 7: 353-364
• 31 Kim SY, Park SM, Hwanqbo M, Lee JR, Byun SH, Ku SK, Cho IJ, Kim SC, Jee SY, Park SJ. Cheongsangbangpung-tang ameliorated the acute inflammatory response via the inhibition of NF-kappaB activation and MAPK phosphorylation. BMC Complement Altern Med 2017; 17: 46
• 32 Torraca V, Mostowy S. Zebrafish infection: from pathogenesis to cell biology. Trends Cell Biol 2018; 28: 143-156
• 33 Mortaz E, Alipoor SD, Adcock IM, Mumby S, Koenderman L. Update on neutrophil function in severe inflammation. Front Immunol 2018; 9: 2171
• 34 Zhou HL, Cao HH, Zheng YR, Lu ZB, Chen YY, Liu DY, Yang HY, Quan JY, Huo CY, Liu JS, Yu LZ. Liang-Ge-San, a classic traditional Chinese medicine formula, attenuates acute inflammation in zebrafish and RAW 264.7 cells. J Ethnopharmacol 2020; 249: 112427
• 35 Pharmacopoeia Committee of Peopleʼs Republic China. Pharmacopoeia of Peopleʼs Republic China. Beijing: Chemical Industry Publishers; 2015: 383-384
• 36 Zhan X, Wu H, Wu H, Wang R, Luo C, Gao B, Chen ZW, Li QL. Metabolites from Bufo gargarizans (Cantor, 1842): A review of traditional uses, pharmacological activity, toxicity and quality control. J Ethnopharmacol 2020; 246: 112178
• 37 Li JJ, Zhang Y, Han LW, Tian QP, He QX, Wang XM, Sun C, Hai J, Liu KC. Tenacissoside H exerts an anti-inflammatory effect by regulating the nf-κb and p38 pathways in zebrafish. Fish Shellfish Immunol 2018; 83: 205-212
• 38 Xiong ST, Wu JJ, Jing J, Huang PP, Li Z, Mei J, Gui JF. Loss of stat3 function leads to spine malformation and immune disorder in zebrafish. Sci Bull 2017; 62: 185-196
• 39 He G, Karin M. NF-κB and STAT3- key players in liver inflammation and cancer. Cell Res 2011; 21: 159-168
• 40 Grivennikov SI, Karin M. Dangerous liaisons: STAT3 and NF-κB collaboration and crosstalk in cancer. Cytokine Growth Factor Rev 2010; 21: 11-19
• 41 Westerfield M. The Zebrafish Book. A Guide for the laboratory Use of Zebrafish (Danio rerio). Eugene: University of Oregon Press; 2000