Akt inhibitor deguelin aggravates inflammation and fibrosis in myocarditis

Document Type: Original Article


1 Department of Forensic Medicine, Xuzhou Medical University, Xuzhou, Jiangsu 221002, China

2 Human anatomy and Histology and Embryology, Xuzhou Medical University, Xuzhou, Jiangsu 221002, China

3 Department of Pharmacology and Experimental Therapeutics, Boston University School of Medicine, Boston, Massachusetts 02118, USA


Objective(s): Myocarditis is characterized by inflammatory cell infiltration in myocardial stroma. Attenuation of tumor necrosis factor (TNF)-α and interleukin (IL)-1β is a reliable mark for improving the prognosis. Protein kinase B (Akt) plays an important role in the development and progression of myocarditis. The specific role of the natural inhibitor of Akt, Deguelin, on myocarditis has not been reported. In this study, we used deguelin to investigate the effects of natural Akt inhibitor on myocarditis in experimental autoimmune myocarditis (EAM) rats.
Materials and Methods: EAM rat models were made by using Lewis rats and Deguelin was injected intraperitoneally on day 3, 6, 9, 12 and 15 after successful modeling. On day 18, rats were sacrificed and the heart weight (HW)/ body weight (BW) ratio were measured. The pathological changes, pathological scores and fibrosis area were evaluated after H.&E. and Masson’s trichrome staining. The mRNA levels of TNF-α and IL-1β were measured by RT-qPCR, while the protein expressions of TNF-α and IL-1β were detected by immunohistochemical staining and Western bolt. The protein expressions of Akt, Akt1, phosphorylated (p-) Akt and nuclear factor (NF)-κB were detected by Western bolt.
Results: We found that the TNF-α and IL-1β levels, inflammatory scores and fibrosis areas were markedly increased after 18 days deguelin administration.
Conclusion: Akt inhibition with deguelin may aggravate myocarditis of EAM rats.


Main Subjects

1. Bunu DM, Timofte CE, Ciocoiu M, Floria M, Tarniceriu CC, Barboi OB, et al. Cardiovascular manifestations of inflammatory bowel disease: Pathogenesis, diagnosis, and preventive strategies. Gastroenterol Res Pract 2019; 2019: 3012509.
2. Delgado-Maroto V, Falo CP, Forte-Lago I, Adan N, Morell M, Maganto-Garcia E, et al. The neuropeptide cortistatin attenuates experimental autoimmune myocarditis via inhibition of cardiomyogenic T cell-driven inflammatory responses. Br J Pharmacol 2017; 174: 267-280.
3. Maya Y, Werner RA, Schütz C, Wakabayashi H, Samnick S, Lapa C, et al. 11C-methionine PET of myocardial inflammation in a rat model of experimental autoimmune myocarditis. J Nucl Med 2016; 57: 1985-1990.
4. Wang H, Yao Q, Zhu S, Zhang G, Wang Z, Li Z, et al. The autopsy study of 553 cases of sudden cardiac death in Chinese adults. Heart Vessels 2014; 29: 486-495.
5. Blauwet LA, Cooper LT. Myocarditis. Prog Cardiovasc Dis 2010; 52: 274-288.
6. Cooper LT. Myocarditis. N Engl J Med 2009; 360: 1526-1538.
7. Zhang Y, Wang X, Yang H, Liu H, Lu Y, Han L, et al. Kinase AKT controls innate immune cell development and function. Immunology 2013; 140: 143-152.
8. Abeyrathna P, Su Y. The critical role of Akt in cardiovascular function. Vascul Pharmacol 2015; 74: 38-48.
9. Song C, Liu B, Xu P, Xie J, Ge X, Zhou Q, et al. Oxidized fish oil injury stress in Megalobrama amblycephala: Evaluated by growth, intestinal physiology, and transcriptome-based PI3K-Akt/NF-κB/TCR inflammatory signaling. Fish Shellfish Immunol 2018; 81:446-455.
10. Ersahin T, Tuncbag N, Cetin-Atalay R. The PI3K/ AKT/ mTOR interactive pathway. Mol Biosyst 2015; 11: 1946-1954..
11. Wang Y, Ma W, Zheng W. Deguelin, a novel anti-tumorigenic agent targeting apoptosis, cell cycle arrest and anti-angiogenesis for cancer chemoprevention. Mol Clin Oncol 2013; 1: 215-219.
12. Martini M, De Santis MC, Braccini L, Gulluni F, Hirsch E, et al. PI3K/ AKT signaling pathway and cancer: an updated review. Ann Med 2014; 46: 372-383.
13. Song Y, Ge W, Cai H. Curcumin protects mice from coxsackievirus B3-induced myocarditis by inhibiting the phosphatidylinositol 3 kinase/ Akt/ nuclear factor-κB pathway. J Cardiovasc Pharmacol Ther 2013; 18: 560-569.
14. Chen Z, Yang L, Liu Y, Tang A, Li X, Zhang J, et al. LY294002 and rapamycin promote coxsackievirus-induced cytopathic effect and apoptosis via inhibition of PI3K/ AKT/mTOR signaling pathway. Mol Cell Biochem 2014; 385: 169-177.
15. Liu HS, Zhang J, Guo JL, Lin CY, Wang ZW. Phosphoinositide 3-kinase inhibitor LY294002 ameliorates the severity of myosin-induced myocarditis in mice. Curr Res Transl Med 2016: 64: 21-27.
16. Gonzalez E, McGraw TE. The Akt kinases: isoform specificity in metabolism and cancer. Cell Cycle 2009;8:2502-2508.
17. Ouyang S, Zeng Q, Tang N, Guo H, Tang R, Yin W, et al. Akt-1 and Akt-2 differentially regulate the development of experimental autoimmune encephalomyelitis by controlling proliferation of thymus-derived regulatory T cells. J Immunol 2019; 202: 1441-1452.
18. Chang H, Li X, Cai Q, Li C, Tian L, Chen J, et al. The PI3K/Akt/mTOR pathway is involved in CVB3-induced autophagy of HeLa cells. Int J Mol Med 2017; 40:182-192.
19. Kang W, Zheng X, Wang P, Guo S. Deguelin exerts anticancer activity of human gastric cancer MGC-803 and MKN-45 cells in vitro. Int J Mol Med 2018; 41:3157-3166.
20. Hsiao YT, Fan MJ, Huang AC, Lien JC, Lin JJ, Chen JC, et al. Deguelin impairs cell adhesion, migration and invasion of human lung cancer cells through the NF-κB signaling pathways. Am J Chin Med 2018; 46: 209-229.
21. Bao Z, Zhang P, Yao Y, Lu G, Tong Z, Yan B, et al. Deguelin attenuates allergic airway inflammation via inhibition of NF-κb pathway in mice. Int J Biol Sci 2017; 13: 492-504.
22. Arms L, Vizzard MA. Role for pAKT in rat urinary bladder with cyclophosphamide (CYP)-induced cystitis. Am J Physiol Renal Physiol 2011; 301: F252-62.
23. Paulus P, Ockelmann P, Tacke S, Karnowski N, Ellinghaus P, Scheller B, et al. Deguelin attenuates reperfusion injury and improves outcome after orthotopic lung transplantation in the rat. PLoS One 2012; 7: e39265.
24. Buss SJ, Riffel JH, Malekar P, Hagenmueller M, Asel C, Zhang M, et al. Chronic Akt blockade aggravates pathological hypertrophy and inhibits physiological hypertrophy. Am J Physiol Heart Circ Physiol 2012; 302: H420-430.
25. Bao Z, Zhang P, Yao Y, Guohua Lu, Zhongkai Tong, Bing Yan, et al. Deguelin attenuates allergic airway inflammation via inhibition of NF-κb pathway in mice. Int J Biol Sci 2017; 13: 492-504.
26. Liu W, Li WM, Gao C, Sun NL. Effects of atorvastatin on the Th1/Th2 polarization of ongoing experimental autoimmune myocarditis in Lewis rats. J Autoimmun 2005; 25: 258-263.
27. Zeng SY, Yang L, Yan QJ, Gao Ling, Lu HQ, Yan PK. Nox1/4 dual inhibitor GKT137831 attenuates hypertensive cardiac remodelling associating with the inhibition of ADAM17-dependent proinflammatory cytokines-induced signalling pathways in the rats with abdominal artery constriction. Biomed Pharmacother 2019;109:1907-1914.
28. Zheng C, Wu SM, Lian H, Lin YZ, Zhuang R, THapa A, Chen QZ, et al. Low-intensity pulsed ultrasound attenuates cardiac inflammation of CVB3-induced viral myocarditis via regulation of caveolin-1 and MAPK pathways. J Cell Mol Med 2019; 23: 1963-1975.
29. Jiang SY, Tian SL, Wu XM, Tao YJ, Jiang DL. Effect of AMP-activated protein kinase activation on cardiac fibroblast proliferation induced by coxsackievirus B3. Exp Ther Med 2016; 11: 2547-2552.
30. Valaperti A, Marty RR, Kania G, Germano D, Mauermann N, Dirnhofer S, et al. CD11b+ monocytes abrogate Th17 CD4+ T cell-mediated experimental autoimmune myocarditis. J Immunol 2008; 180: 2686-2695.
31. Esfandiarei M, Suarez A, Amaral A, Si X, Rahmani M, Dedhar S, et al. Novel role for integrin-linked kinase in modulation of coxsackievirus B3 replication and virus-induced cardiomyocyte injury. Circ Res 2006; 99: 354-361.
32. Watanabe K, Sukumaran V, Veeraveedu PT, Thandavarayan RA, Gurusamy N, Ma M, et al. Regulation of inflammation and myocardial fibrosis in experimental autoimmune myocarditis. Inflamm Allergy Drug Targets 2011; 10: 218-225.
33. Lee WS, Erdelyi K, Matyas C, Mukhopadhyay P, Varga ZV, Liaudet L, et al. Cannabidiol limits T cell-mediated chronic autoimmune myocarditis: implications to autoimmune disorders and organ transplantation. Mol Med 2016; 22:136-146.
34. Wu B, Li J, Ni H, Mukhopadhyay P, Varga ZV, Liaudet L, et al. TLR4 activation promotes the progression of experimental autoimmune myocarditis to dilated cardiomyopathy by inducing mitochondrial dynamic imbalance. Oxid Med Cell Longev 2018; 3181278.
35. Nana-Leventaki E, Nana M, Poulianitis N, Sampaziotis D, Perrea D, Sanoudou D, et al. Cardiosphere-derived cells attenuate inflammation, preserve systolic function, and prevent adverse remodeling in rat hearts with experimental autoimmune myocarditis. J Cardiovasc Pharmacol Ther 2018; 24: 70-77.
36. Zhang JY, Yang Z, Fang K, Shi ZL, Ren DH, Sun J. Oleuropein prevents the development of experimental autoimmune myocarditis in rats. Int Immunopharmacol 2017; 48: 187-195.
37. Zhang H, Song Y. Glycyrrhizin administration ameliorates coxsackievirus B3-induced myocarditis in mice. Am J Med Sci 2012; 344: 206-210.
38. Noel R. Rose. Critical cytokine pathways to cardiac inflammation. J Interferon Cytokine Res 2011; 31: 705-710.
39. Reyes-García J, Flores-Soto E, Solís-Chagoyán H, Sommer B, Díaz-Hernández Verónica, Luz María García-Hernández, et al. Tumor necrosis factor alpha inhibits L-type Ca(2+) channels in sensitized guinea pig airway smooth muscle through ERK 1/2 pathway. Mediators Inflamm 2016; 2016: 5972302.
40. Duncan DJ, Yang Z, Hopkins PM, Steele DS, Harrison SM. TNF-alpha and IL-1beta increase Ca2+ leak from the sarcoplasmic reticulum and susceptibility to arrhythmia in rat ventricular myocytes. Cell calcium 2010; 47: 378-386.
41. Liu T, Zhang L, Joo D, Sun SC. NF-κB signaling in inflammation. Signal Transduct Target Ther 2017; 2: e17023.
42. Gui J, Yue Y, Chen R, Xu W, Xiong S. A20 (TNFAIP3) alleviates CVB3-induced myocarditis via inhibiting NF-κB signaling. PloS one 2012; 7: e46515.
43. Metelev VG, Kubareva EA, Oretskaya TS. Regulation of activity of transcription factor NF-κB by synthetic oligonucleotides. Biochemistry (Mosc) 2013; 78: 867-878.
44. Henao-Martínez AF, Agler AH, Watson AM, Hennessy C, Davidson E, Demos-Davies K. AKT network of genes and impaired myocardial contractility during murine acute Chagasic myocarditis. Am J Trop Med Hyg 2015; 92: 523-529.
45. Henao-Martínez AF, Parra-Henao G. Murine heart gene expression during acute Chagasic myocarditis. Genom Data 2015; 4: 76-77.
46. He H, Chang X, Gao J, Zhu L, Miao M, Yan T. Salidroside mitigates sepsis-induced myocarditis in rats by regulating IGF-1 / PI3K / Akt / GSK-3β signaling. Inflammation 2015; 38: 2178-2184.
47. Lee HY. Molecular mechanisms of deguelin-induced apoptosis in transformed human bronchial epithelial cells. Biochem Pharmacol 2004; 68: 1119-1124.
48. Baba Y, Fujii M, Maeda T, Suzuki A, Yuzawa S, Kato Y. Deguelin induces apoptosis by targeting both EGFR-Akt and IGF1R-Akt pathways in head and neck squamous cell cancer cell lines. Biomed Res Int 2015; 2015: 657179.
49. Day TF, Kallakury B, Ross JS, Voronel O, Vaidya S, Sheehan CE, et al. Dual targeting of EGFR and IGF1R in the TNFAIP8 knockdown non-small cell lung cancer cells. Mol Cancer Res 2019; 17: 1207-1219.
50. Liu YP, Lee JJ, Lai TC, Lee CH, Hsiao YW, Chen PS, et al. Suppressive function of low-dose deguelin on the invasion of oral cancer cells by downregulating tumor necrosis factor alpha-induced nuclear factor-kappa B signaling. Head Neck 2016; 38 Suppl 1: E524-34.
51. Zhao L, Yang XR, Han X. MicroRNA-146b induces the PI3K / Akt / NF-κB signaling pathway to reduce vascular inflammation and apoptosis in myocardial infarction by targeting PTEN. Exp Ther Med 2019; 17: 1171-1181.
52. Wang A, Wang W, Chen Y, Ma F, Wei X, Bi Y. Deguelin induces PUMA-mediated apoptosis and promotes sensitivity of lung cancer cells (LCCs) to doxorubicin (Dox). Mol Cell Biochem 2018; 442: 177-186.