Vitexin ameliorates GCDC-induced hepatocyte injury through SIRT6 and JAK2/STAT3 pathways

Document Type : Original Article


Department of Pediatric Surgery, Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, 050000, China


Objective(s): Vitexin, a natural flavonoid, is commonly found in many foods and traditional herbal medicines and has clear health benefits. However, the role of vitexin in cholestasis is presently unclear. This study investigated whether vitexin mitigated glycochenodeoxycholate (GCDC)-induced hepatocyte injury and further elucidated the underlying mechanisms.
Materials and Methods: A cell counting kit-8 (CCK-8) assay was conducted to evaluate cell viability. The mitochondrial membrane potential (MMP, Δψm), reactive oxygen species (ROS) levels, and apoptosis rate of hepatocytes exposed to GCDC were detected by flow cytometry (FCM). We then measured the cytoprotective effects of vitexin against oxidative stress. The molecular signaling pathway was further investigated by using Western blotting and signaling pathway inhibitors.
Results: Here, we showed that vitexin increased cell viability and reduced cell apoptosis, necroptosis, and oxidative stress in a dose-dependent manner in GCDC-treated hepatocytes. In addition, by using selective inhibitors, we further confirmed that inhibition of the JAK2/STAT3 pathway by vitexin was mediated by prolonged activation of Sirtuin 6 (SIRT6).
Conclusion: Vitexin attenuated GCDC-induced hepatocyte injury via SIRT6 and the JAK2/STAT3 pathways.


1. Hu G, He P, Liu Z, Chen Q, Zheng B, Zhang Q. Diagnosis of ABCB11 gene mutations in children with intrahepatic cholestasis using high resolution melting analysis and direct sequencing. Molecular medicine reports 2014; 10:1264-1274.
2. Koh H, Park G-S, Shin S-M, Park CE, Kim S, Han SJ, et al. Mitochondrial mutations in cholestatic liver disease with biliary atresia. Scientific Reports 2018; 8:905-905.
3. Ibrahim S, Dayoub R, Saberi V, Buchner M, Melter M, Weiss TS. Augmenter of Liver Regeneration (ALR) regulates bile acid synthesis and attenuates bile acid-induced apoptosis via glycogen synthase kinase-3β (GSK-3β) inhibition. Exp Cell Res 2020; 397:112343.
4. Ganjam GK, Terpolilli NA, Diemert S, Eisenbach I, Hoffmann L, Reuther C, et al. Cylindromatosis mediates neuronal cell death in vitro and in vivo. Cell death and differentiation 2018; 25:1394-1407.
5. Afonso MB, Rodrigues CMP. Necrosome Formation and Necroptosis in Experimental Cholestasis. Methods Mol Biol 2019; 1981:149-162.
6. Li J, Yu D, Chen S, Liu Y, Shi J, Zhang J, et al. Sirt6 opposes glycochenodeoxycholate-induced apoptosis of biliary epithelial cells through the AMPK/PGC-1α pathway. Cell Biosci 2020; 10:43-54.
7. Yu T, Wang L, Lee H, O’Brien DK, Bronk SF, Gores GJ, et al. Decreasing mitochondrial fission prevents cholestatic liver injury. J Biol Chem 2014; 289:34074-34088.
8. Deng Y, Zhang J, Sun X, Ma G, Luo G, Miao Z, et al. miR-132 improves the cognitive function of rats with Alzheimer’s disease by inhibiting the MAPK1 signal pathway. Exp Ther Med 2020; 20:159-159.
9. Hu XK, Rao SS, Tan YJ, Yin H, Luo MJ, Wang ZX, et al. Fructose-coated Angstrom silver inhibits osteosarcoma growth and metastasis via promoting ROS-dependent apoptosis through the alteration of glucose metabolism by inhibiting PDK. Theranostics 2020; 10:7710-7729.
10. Yoon JJ, Son CO, Kim HY, Han BH, Lee YJ, Lee HS, et al. Betulinic acid protects dox-triggered cardiomyocyte hypertrophy response through the GATA-4/Calcineurin/NFAT pathway. Molecules 2020; 26:53-66.
11. Hao L, Bang IH, Wang J, Mao Y, Yang JD, Na SY, et al. ERRγ suppression by Sirt6 alleviates cholestatic liver injury and fibrosis. JCI Insight 2020; 5.
12. Zhang J, Li Y, Liu Q, Huang Y, Li R, Wu T, et al. Sirt6 alleviated liver fibrosis by deacetylating conserved lysine 54 on Smad2 in hepatic stellate cells. Hepatology 2021;73:1140-1157.
13. Zhong X, Huang M, Kim HG, Zhang Y, Chowdhury K, Cai W, et al. SIRT6 protects against liver fibrosis by deacetylation and suppression of SMAD3 in hepatic stellate cells. Cell Mol Gastroenterol Hepatol 2020; 10:341-364.
14. Zhang Y, Biao Y, Chu X, Hao L, Shi C, Liu Y, et al. Protective effect of Chushizi (Fructus Broussonetiae) on acetaminophen-induced rat hepatitis by inhibiting the Toll-like receptor 3/c-Jun N-terminal kinase/c-jun/c-fos/janus protein tyrosine kinase/activators of transcription 3 pathway. J Tradit Chin Med 2020; 40:965-973.
15. An X, Liu J, Li Y, Dou Z, Li N, Suo Y, et al. Chemerin/CMKLR1 ameliorates nonalcoholic steatohepatitis by promoting autophagy and alleviating oxidative stress through the JAK2-STAT3 pathway. Peptides 2021; 135:170422.
16. Lee CH, Choi Y, Cho H, Bang IH, Hao L, Lee SO, et al. Histone deacetylase 8 inhibition alleviates cholestatic liver injury and fibrosis. Biochem Pharmacol 2021; 183:114312.
17. Lyu Z, Cao J, Wang J, Lian H. Protective effect of vitexin reduces sevoflurane-induced neuronal apoptosis through HIF-1α, VEGF and p38 MAPK signaling pathway in vitro and in newborn rats. Exp Ther Med 2018; 15:3117-3123.
18. Zhao Q, Yang R, Wang J, Hu DD, Li F. PPARα activation protects against cholestatic liver injury. Sci Rep 2017; 7:9967-9979.
19. Gao L, Lv G, Guo X, Jing Y, Han Z, Zhang S, et al. Activation of autophagy protects against cholestasis-induced hepatic injury. Cell Biosci 2014; 4:47-56.
20. Greimel T, Jahnel J, Pohl S, Strini T, Tischitz M, Meier-Allard N, et al. Bile acid-induced tissue factor activity in hepatocytes correlates with activation of farnesoid X receptor. Lab Invest 2021; 101:1394-1402.
21. Arenas F, Hervias I, Uriz M, Joplin R, Prieto J, Medina JF. Combination of ursodeoxycholic acid and glucocorticoids up-regulates the AE2 alternate promoter in human liver cells. J Clin Invest 2008; 118:695-709.
22. Jiang H, Cheng S-T, Ren J-H, Ren F, Yu H-B, Wang Q, et al. SIRT6 Inhibitor, OSS_128167 restricts hepatitis B virus transcription and replication through targeting transcription factor peroxisome proliferator-activated receptors α. Front Pharmacol 2019; 10:1270-1270.
23. Yu T, Wang L, Lee H, O’Brien DK, Bronk SF, Gores GJ, et al. Decreasing mitochondrial fission prevents cholestatic liver injury. The Journal of biological chemistry 2014; 289:34074-34088.
24. Kim JY, Jang HJ, Leem J, Kim GM. Protective effects of bee venom-derived phospholipase A(2) against cholestatic liver disease in mice. Biomedicines 2021; 992-1004.
25. Gonzalez B, Fisher C, Rosser BG. Glycochenodeoxycholic acid (GCDC) induced hepatocyte apoptosis is associated with early modulation of intracellular PKC activity. Mol Cell Biochem 2000; 207:19-27.
26. Li J, Yu D, Chen S, Liu Y, Shi J, Zhang J, et al. Sirt6 opposes glycochenodeoxycholate-induced apoptosis of biliary epithelial cells through the AMPK/PGC-1α pathway. Cell Biosci 2020; 10:43-54.
27. Afonso MB, Rodrigues PM, Simão AL, Ofengeim D, Carvalho T, Amaral JD, et al. Activation of necroptosis in human and experimental cholestasis. Cell Death Dis 2016; 7:e2390-e2390.
28. Yoon JJ, Son CO, Kim HY, Han BH, Lee YJ, Lee HS, et al. Betulinic acid protects DOX-triggered cardiomyocyte hypertrophy response through the GATA-4/calcineurin/NFAT pathway. Molecules (Basel, Switzerland) 2020; 26:53.
29. Hohenester S, Kanitz V, Kremer AE, Paulusma CC, Wimmer R, Kuehn H, et al. Glycochenodeoxycholate promotes liver fibrosis in mice with hepatocellular cholestasis. Cells 2020; 9.
30. Wang K, Wang M, Gannon M, Holterman A. Growth hormone mediates its protective effect in hepatic apoptosis through Hnf6. PLoS One 2016; 11:e0167085.
31. Molaei P, Vaseghi S, Entezari M, Hashemi M, Nasehi M. The effect of neuroaid (MLC901) on cholestasis-induced spatial memory impairment with respect to the expression of BAX, BCL-2, BAD, PGC-1α and TFAM genes in the hippocampus of male wistar rats. Neurochem Res 2021; 46:2154-2166.
32. Bai L, Kong M, Duan Z, Liu S, Zheng S, Chen Y. M2-like macrophages exert hepatoprotection in acute-on-chronic liver failure through inhibiting necroptosis-S100A9-necroinflammation axis. Cell Death Dis 2021; 12:93-105.
33. Afonso MB, Rodrigues PM, Simão AL, Gaspar MM, Carvalho T, Borralho P, et al. miRNA-21 ablation protects against liver injury and necroptosis in cholestasis. Cell Death Differ 2018; 25:857-872.
34. Qiu LZ, Yue LX, Ni YH, Zhou W, Huang CS, Deng HF, et al. Emodin-induced oxidative inhibition of mitochondrial function assists BiP/IRE1α/CHOP signaling-mediated ER-related apoptosis. Oxid Med Cell Longev 2021; 2021:8865813.
35. Ebrahimi R, Sepand MR, Seyednejad SA, Omidi A, Akbariani M, Gholami M, et al. Ellagic acid reduces methotrexate-induced apoptosis and mitochondrial dysfunction via up-regulating Nrf2 expression and inhibiting the IĸBα/NFĸB in rats. Daru 2019; 27:721-733.
36. Zhong M, Che L, Du M, Liu K, Wang D. Desflurane protects against liver ischemia/reperfusion injury via regulating miR-135b-5p. J Chin Med Assoc 2021; 84:38-45.