Cepharanthine hydrochloride alleviates herpesvirus encephalitis by inhibiting Nrf2 degradation

Qiongzhen Zeng; Weixiangmin Zou; Jiaqi Wu; Menghe Li; Zexu Wang; Caiwenjie La; Cuifang Ye; Zhe Ren; Yifei Wang; Kai Zheng

doi:10.1016/j.virs.2026.03.016

April 2026

Citation: Qiongzhen Zeng, Weixiangmin Zou, Jiaqi Wu, Menghe Li, Zexu Wang, Caiwenjie La, Cuifang Ye, Zhe Ren, Yifei Wang, Kai Zheng. Cepharanthine hydrochloride alleviates herpesvirus encephalitis by inhibiting Nrf2 degradation .VIROLOGICA SINICA, 2026, 41(2) : 392-403. http://dx.doi.org/10.1016/j.virs.2026.03.016

Cepharanthine hydrochloride alleviates herpesvirus encephalitis by inhibiting Nrf2 degradation

Qiongzhen Zeng ^a,b ,
Weixiangmin Zou ^a,c ,
Jiaqi Wu ^a,c ,
Menghe Li ^a,c ,
Zexu Wang ^a,c ,
Caiwenjie La ^a,c ,
Cuifang Ye ^a ,
Zhe Ren ^{a,c
,,} ,
Yifei Wang ^{a,c
,,} ,
Kai Zheng ^{d
,,}

a. Department of Cell Biology, College of Life Science and Technology, Jinan University, Guangzhou 510632, China;
b. Guangdong Provincial Clinical Research Center for Geriatrics, Shenzhen Clinical Research Center for Geriatrics, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University, The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen 518020, China;
c. Institute of Biomedicine, Guangdong Province Key Laboratory of Bioengineering Medicine, Key Laboratory of Innovative Technology Research on Natural Products and Cosmetics Raw Materials, Jinan University, Guangzhou 510632, China;
d. School of Pharmacy, Shenzhen University Medical School, Shenzhen University, Shenzhen 518055, China

Corresponding author: Zhe Ren, rz62@163.com
Yifei Wang, twang-yf@163.com
Kai Zheng, zhengk@szu.edu.cn
Received Date: 09 October 2025
Accepted Date: 30 March 2026
Available online: 01 April 2026

Abstract

Herpes simplex virus type 1 (HSV-1) infection can induce herpes simplex encephalitis (HSE), a life-threatening neurological disorder characterized by active viral replication within the central nervous system accompanied by excessive neuroinflammatory responses. Cepharanthine hydrochloride (CH) is a natural bisbenzylisoquinoline alkaloid with diverse pharmacological activities. CH was evaluated for its antiviral and anti-inflammatory activities against HSV-1 infection. In microglia, CH markedly inhibited viral replication and suppressed STING-NF-κB signaling, thereby reducing HSV-1-associated neuroinflammation. During HSV-1 infection, CH binds to Nrf2, disrupting Keap1-Nrf2 interaction and subsequent ubiquitination. This hindered Nrf2 degradation and attenuated STING activation. In an HSE mouse model, CH significantly reduced viral loads in brain tissue, improved survival rates, prevented weight loss, and alleviated neurological symptoms. Furthermore, CH promoted the accumulation of Nrf2 and inhibited the STING-NF-κB signaling pathway in vivo. Collectively, these results indicate that CH represents a potential antiviral strategy for HSE.
- Herpes simplex encephalitis (HSE)
- , Neuroinflammation
- , Cepharanthine hydrochloride (CH)
- , Nrf2
- , STING-NF-κB pathway

Electronic Supplementary Material

10.1016j.virs.2026.03.016-ESM.docx
References
1. Bellezza, I., Giambanco, I., Minelli, A.,Donato, R., 2018. Nrf2-Keap1 signaling in oxidative and reductive stress. Biochim. Biophys. Acta Mol. Cell Res., 1865, 721-733.
2. Benarroch, E.E., 2017. Nrf2, cellular redox regulation, and neurologic implications. Neurology, 88, 1942-1950.
3. Bradshaw, M.J.,Venkatesan, A., 2016. Herpes simplex Virus-1 encephalitis in adults: pathophysiology, diagnosis, and management. Neurotherapeutics, 13, 493-508.
4. Chen, G., Wen, D., Shen, L., Feng, Y., Xiong, Q., Li, P.,Zhao, Z., 2023. Cepharanthine exerts antioxidant and anti-inflammatory effects in lipopolysaccharide (LPS)-induced macrophages and DSS-induced colitis mice. Molecules, 28.
5. Chen, K., Lai, C., Su, Y., Bao, W.D., Yang, L.N., Xu, P.P.,Zhu, L.Q., 2022. cGAS-STING-mediated IFN-I response in host defense and neuroinflammatory diseases. Curr. Neuropharmacol., 20, 362-371.
6. Conrady, C.D., Zheng, M., Van Rooijen, N., Drevets, D.A., Royer, D., Alleman, A.,Carr, D.J., 2013. Microglia and a functional type I IFN pathway are required to counter HSV-1-driven brain lateral ventricle enlargement and encephalitis. J. Immunol., 190, 2807-2817.
7. Crisman, E., Duarte, P., Dauden, E., Cuadrado, A., Rodríguez-Franco, M.I., López, M.G., León, R., 2023. KEAP1-NRF2 protein-protein interaction inhibitors: Design, pharmacological properties and therapeutic potential. Med Res Rev 43, 237–287.
8. Daskou, M., Fotooh Abadi, L., Gain, C., Wong, M., Sharma, E., Kombe Kombe, A.J., Nanduri, R.,Kelesidis, T., 2023. The role of the NRF2 pathway in the pathogenesis of viral respiratory infections. Pathogens, 13.
9. Dong, Z., Chang, X., Luo, X., Li, H., Deng, M., Huang, Z., Chen, T., Chen, Y., Sun, B., Wu, Y., Wu, R., Wu, Q., Zhu, J.,Zheng, J., 2025. Integration of network pharmacology, transcriptomics, and experimental verification to investigate the mechanism of action of cepharanthine hydrochloride against prostate cancer. Sci. Rep., 15, 18115.
10. Guo, Y., Ou, C., Zhang, N., Liu, Q., Xiong, K., Yu, J., Cheng, H., Chen, L., Ma, M., Xu, J., Wu, J., 2023. Roflumilast attenuates neuroinflammation post retinal ischemia/reperfusion injury by regulating microglia phenotype via the Nrf2/STING/NF-κB pathway. Int Immunopharmacol 124, 110952.
11. Guo, W., Yang, Y., Liu, G., Zhao, J., Zhang, Y., Li, Y., Yang, B., Zhu, X., Li, D., Qin, X., Zhang, P., Yang, Z., Guo, W., Kong, D.,Zhang, W., 2025. The anti-neuroinflammatory effects of cepharanthine in uric acid-induced neuroinflammation. J. Ethnopharmacol., 342, 119409.
12. Huang, Z., Li, S., Zhong, L., Su, Y., Li, M., Wang, X., Wang, Z., Wang, Z., Ye, C., Ren, Z., Wang, X., Zeng, Q., Zheng, K.,Wang, Y., 2024. Effect of resveratrol on herpesvirus encephalitis: evidences for its mechanisms of action. Phytomedicine, 127, 155476.
13. Itoh, K., Wakabayashi, N., Katoh, Y., Ishii, T., Igarashi, K., Engel, J.D., Yamamoto, M., 1999. Keap1 represses nuclear activation of antioxidant responsive elements by Nrf2 through binding to the amino-terminal Neh2 domain. Genes Dev 13, 76–86.
14. Katzilieris-Petras, G., Lai, X., Rashidi, A.S., Verjans, G., Reinert, L.S.,Paludan, S.R., 2022. Microglia activate early antiviral responses upon Herpes simplex virus 1 entry into the brain to counteract development of encephalitis-like disease in mice. J. Virol., 96, e0131121.
15. Kim, B., Kim, Y.S., Li, W., Kwon, E.B., Chung, H.S., Go, Y.,Choi, J.G., 2024. Ginsenoside Rg5, a potent agonist of Nrf2, inhibits HSV-1 infection-induced neuroinflammation by inhibiting oxidative stress and NF-κB activation. J Ginseng Res, 48, 384-394.
16. Kou, B., Zhang, Y., Zhang, W., Zhang, J.,Jiang, R., 2025. STING regulates porphyromonas gingivalis lipopolysaccharide-induced pyroptosis and inflammatory response through the NF-κB/NLRP3 signaling pathway in human gingival fibroblasts. Arch. Oral Biol., 173, 106197.
17. Li, N., Wang, J., Zang, X., Wang, Z., Zhang, T., Zhao, B., Miao, J.,Lin, Z., 2021. H(2)S probe CPC inhibits autophagy and promotes apoptosis by inhibiting glutathionylation of Keap1 at Cys434. Apoptosis, 26, 111-131.
18. Liu, Y., Jesus, A.A., Marrero, B., Yang, D., Ramsey, S.E., Sanchez, G.a.M., Tenbrock, K., Wittkowski, H., Jones, O.Y., Kuehn, H.S., et al., 2014. Activated STING in a vascular and pulmonary syndrome. N. Engl. J. Med., 371, 507-518.
19. Lo, S.C., Li, X., Henzl, M.T., Beamer, L.J.,Hannink, M., 2006. Structure of the Keap1:Nrf2 interface provides mechanistic insight into Nrf2 signaling. EMBO J., 25, 3605-3617.
20. Marcocci, M.E., Napoletani, G., Protto, V., Kolesova, O., Piacentini, R., Li Puma, D.D., Lomonte, P., Grassi, C., Palamara, A.T.,De Chiara, G., 2020. Herpes simplex Virus-1 in the brain: the dark side of a sneaky infection. Trends Microbiol., 28, 808-820.
21. Motwani, M., Pesiridis, S.,Fitzgerald, K.A., 2019. DNA sensing by the cGAS-STING pathway in health and disease. Nat. Rev. Genet., 20, 657-674.
22. Olagnier, D., Brandtoft, A.M., Gunderstofte, C., Villadsen, N.L., Krapp, C., Thielke, A.L., Laustsen, A., Peri, S., Hansen, A.L., Bonefeld, L., et al., 2018. Nrf2 negatively regulates STING indicating a link between antiviral sensing and metabolic reprogramming. Nat. Commun., 9, 3506.
23. Paul, B.D., Snyder, S.H.,Bohr, V.A., 2021. Signaling by cGAS-STING in neurodegeneration, neuroinflammation, and aging. Trends Neurosci., 44, 83-96.
24. Piret, J.,Boivin, G., 2020. Immunomodulatory strategies in Herpes simplex virus encephalitis. Clin. Microbiol. Rev., 33.
25. Qin, D., Song, H., Wang, C., Ma, X., Fu, Y., Zhao, C., Zhao, W., Zhang, L., Zhang, W., 2024. ZC3HAV1 facilitates STING activation and enhances inflammation. Commun Biol 7, 1418.
26. Reinert, L.S., Lopusna, K., Winther, H., Sun, C., Thomsen, M.K., Nandakumar, R., Mogensen, T.H., Meyer, M., Vaegter, C., Nyengaard, J.R., Fitzgerald, K.A.,Paludan, S.R., 2016. Sensing of HSV-1 by the cGAS-STING pathway in microglia orchestrates antiviral defence in the CNS. Nat. Commun., 7, 13348.
27. Reinert, L.S., Rashidi, A.S., Tran, D.N., Katzilieris-Petras, G., Hvidt, A.K., Gohr, M., Fruhwürth, S., Bodda, C., Thomsen, M.K., Vendelbo, M.H., et al., 2021. Brain immune cells undergo cGAS/STING-dependent apoptosis during herpes simplex virus type 1 infection to limit type I IFN production. J Clin Invest 131.
28. Rogosnitzky, M., Danks, R., 2011. Therapeutic potential of the biscoclaurine alkaloid, cepharanthine, for a range of clinical conditions. Pharmacol Rep 63, 337–347.
29. Sun, X., Xie, Z., Hu, B., Zhang, B., Ma, Y., Pan, X., Huang, H., Wang, J., Zhao, X., Jie, Z., Shi, P., Chen, Z., 2020. The Nrf2 activator RTA-408 attenuates osteoclastogenesis by inhibiting STING dependent NF-κb signaling. Redox Biol 28, 101309.
30. Uruno, A., Yamamoto, M., 2023. The KEAP1-NRF2 System and Neurodegenerative Diseases. Antioxid Redox Signal 38, 974–988.
31. Wang, E., Huang, X., Ye, Y., Zou, S., Chen, G., Yang, L., Fraser, N.W., Bao, F., Zhou, J., Cao, X., 2023. Persistent inflammation and neuronal loss in the mouse brain induced by a modified form of attenuated herpes simplex virus type I. Virol Sin 38, 108–118.
32. Wang, H., Jia, X., Zhang, M., Cheng, C., Liang, X., Wang, X., Xie, F., Wang, J., Yu, Y., He, Y., Dong, Q., Wang, Y., Xu, A., 2023. Isoliquiritigenin inhibits virus replication and virus-mediated inflammation via NRF2 signaling. Phytomedicine 114, 154786.
33. Wang, Y., Luo, W., Wang, X., Ma, Y., Huang, L., Wang, Y., 2022. MAMDC2, a gene highly expressed in microglia in experimental models of Alzheimers Disease, positively regulates the innate antiviral response during neurotropic virus infection. J Infect 84, 187–204.
34. Wang, Y., Su, G., Huang, Z., Fan, J.,Wang, Y., 2019. Cepharanthine hydrochloride degrades polyglutamine-expanded androgen receptor proteins through an autophagy pathway in neuron cells. Eur. J. Pharmacol., 861, 172534.
35. Wang, Z., Zou, W., Zeng, Q., Song, X., Li, M., Pang, J., Zhu, H., La, C., Wang, X., Wang, Y.,Zheng, K., 2025. Novel Hsp90α inhibitor inhibits HSV-1 infection by suppressing the Akt/β-catenin pathway. Int. J. Antimicrob. Agents, 65, 107448.
36. Wu, S., Wang, S., Lin, X., Yang, S., Ba, X., Xiong, D., Xiao, L., Li, R., 2024. Lanatoside C inhibits herpes simplex virus 1 replication by regulating NRF2 distribution within cells. Phytomedicine 124, 155308.
37. Xu, L.T., Han, Q.T., Xu, Z.P., Wang, X.N., Zhang, H.,Shen, T., 2025. Tussilagone attenuated cigarette smoke-induced chronic obstructive pulmonary disease through regulating Nrf2 and NF-κB/NLRP3 inflammasome via directly targeting cysteine 434 of KEAP1. J. Adv. Res., 10.1016/j.jare.2025.07.019.
38. Xu, X.Q., Xu, T., Ji, W., Wang, C., Ren, Y., Xiong, X., Zhou, X., Lin, S.H., Xu, Y., Qiu, Y., 2023. Herpes Simplex Virus 1-Induced Ferroptosis Contributes to Viral Encephalitis. mBio 14, e0237022.
39. Yamashiro, L.H., Wilson, S.C., Morrison, H.M., Karalis, V., Chung, J.J., Chen, K.J., Bateup, H.S., Szpara, M.L., Lee, A.Y., Cox, J.S.,Vance, R.E., 2020. Interferon-independent STING signaling promotes resistance to HSV-1 in vivo. Nat. Commun., 11, 3382.
40. Yin, Q., Tian, Y., Kabaleeswaran, V., Jiang, X., Tu, D., Eck, M.J., Chen, Z.J.,Wu, H., 2012. Cyclic di-GMP sensing via the innate immune signaling protein STING. Mol. Cell, 46, 735-745.
41. Yin, C., Yang, P., Xiao, Q., Sun, P., Zhang, X., Zhao, J., Hu, X., Shan, C., 2024. Novel antiviral discoveries for Japanese encephalitis virus infections through reporter virus-based high-throughput screening. J Med Virol 96, e29382.
42. Zhang, S.Y.,Casanova, J.L., 2024. Genetic defects of brain immunity in childhood herpes simplex encephalitis. Nature, 635, 563-573.
43. Zhou, Y.B., Wang, Y.F., Zhang, Y., Zheng, L.Y., Yang, X.A., Wang, N., Jiang, J.H., Ma, F., Yin, D.T., Sun, C.Y.,Wang, Q.D., 2012. In vitro activity of cepharanthine hydrochloride against clinical wild-type and lamivudine-resistant hepatitis B virus isolates. Eur. J. Pharmacol., 683, 10-15.
44. Zou, J., Zheng, Z., Ye, W., Jin, M., Yang, P., Little, P.J., Wang, J.,Liu, Z., 2024. Targeting the smooth muscle cell KEAP1-Nrf2-STING axis with pterostilbene attenuates abdominal aortic aneurysm. Phytomedicine, 130, 155696.
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