Expression of caspase-8 in non-alcoholic fatty liver disease related to ligature-induced periodontitis in rats

Authors

  • Felipe Rodolfo Pereira da Silva Universidade Federal do Pará (UFPA), Faculdade de Medicina
  • Victor Brito Dantas Martins Universidade Federal do Delta do Parnaíba (UFDPar), Laboratório de Análises e Preparo Histológico (LAPHis)
  • Hélio Mateus Silva Nascimento Universidade Federal do Delta do Parnaíba (UFDPar), Laboratório de Análises e Preparo Histológico (LAPHis)
  • Paulo Roberto Carneiro Gomes Universidade Federal do Delta do Parnaíba (UFDPar), Laboratório de Análises e Preparo Histológico (LAPHis)
  • Nikaely Brandão Barbosa Universidade Federal do Delta do Parnaíba (UFDPar), Laboratório de Análises e Preparo Histológico (LAPHis)
  • Vinícius da Silva Caetano Universidade Federal do Delta do Parnaíba (UFDPar), Laboratório de Análises e Preparo Histológico (LAPHis)
  • Ivana Márcia Alves Diniz Universidade Federal de Minas Gerais (UFMG), Faculdade de Odontologia, Departamento de Dentística Restaurador
  • Karine Duarte da Silva Universidade Federal de Minas Gerais (UFMG), Faculdade de Odontologia, Departamento de Patologia e Cirurgia Oral
  • Any Carolina Cardoso Guimarães Vasconcelos Instituto de Educação do Vale do Parnaíba (IESVAP - Afya), Faculdade de Medicina
  • Daniel Fernando Pereira Vasconcelos Universidade Federal do Delta do Parnaíba (UFDPar), Laboratório de Análises e Preparo Histológico (LAPHis)

DOI:

https://doi.org/10.1590/1678-7765-2025-0717

Keywords:

Apoptosis, Periodontal Disease, Liver dysfunction, Oxidative stress

Abstract

Aim  This study aimed to evaluate the oral and hepatic changes, as well as the expression of caspase-8 in liver tissue damage, both associated with ligature-induced periodontitis in rats. Methodology  A total of 16 female Wistar rats were divided into two groups (n=8): one received ligature around the first lower molar to promote periodontitis, and the other was the control group with no ligature. After the period of induction of the disease the animals were evaluated by clinical measures for periodontitis, euthanized, and the samples of gingival tissue, hepatic tissue, and serum were collected to the measurement of biomarkers for inflammation (myeloperoxidase), oxidative stress (glutathione, malonaldehyde, and nitrate [NO3]), liver damage and histopathological evaluation with immunohistochemistry for caspase-8. The data were expressed as mean and standard deviation for data with normal distribution or median and interquartile range for data with non-normal distribution. We used ANOVA followed by the Student-Newman-Keuls test for multiple comparisons of normally distributed data, and the Kruskal-Wallis test followed by Dunn's test for non-normally distributed data. The statistical tests were performed with GraphPad Prism Software (version 5.0), in which a p-value <0.05 was considered significant. Results  Our study demonstrates that the group with ligature-induced periodontitis showed increased measurement of periodontal destruction, local and systemic biomarkers of inflammation as well as liver damage which we observed several hepatocytes with loss of conformation and steatosis in the periodontitis group. The histopathological evaluation evidenced the periodontitis-related steatosis and higher expression of Caspase-8 in comparison with the control group (p<0.0006). Conclusion  Our study demonstrates the high expression of Caspase-8 in liver damage related to ligature-induced periodontitis in rats.

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References

1- Silva FR, Pessoa LS, Shin JI, Alves EH, Koga RS, Smith CV, et al. Polymorphisms in the interleukin genes and chronic periodontitis: a field synopsis and revaluation by Bayesian approaches. Cytokine. 2021;138:155361. doi: 10.1016/j.cyto.2020.155361

» https://doi.org/10.1016/j.cyto.2020.155361

2- Dai X, Dai M, Liang Y, Li X, Zhao W. Global burden and trends of oral disorders among adolescent and young adult (10-24 years old) from 1990 to 2021. BMC Oral Health. 2025;25(1):486. doi: 10.1186/s12903-025-05864-z

» https://doi.org/10.1186/s12903-025-05864-z

3- Zhang X, Wang X, Wu J, Wang M, Hu B, Qu H, et al. The global burden of periodontal diseases in 204 countries and territories from 1990 to 2019. Oral Dis. 2024;30(2):754-68. doi: 10.1111/odi.14436

» https://doi.org/10.1111/odi.14436

4- Isola G, Polizzi A, Serra S, Boato M, Sculean A. Relationship between periodontitis and systemic diseases: a bibliometric and visual study. Periodontol 2000. 2025;98(1):228-40. doi: 10.1111/prd.12621

» https://doi.org/10.1111/prd.12621

5- Galeno JG, França LF, Silva FR, Alves EH, Lenardo DD, Nascimento HM, et al. Renal alterations caused by ligature-induced periodontitis persist after ligature removal in rats. J Periodontal Res. 2021;56(2):306-13. doi: 10.1111/jre.12822

» https://doi.org/10.1111/jre.12822

6- Ayala KN, Caetano VD, Vasconcelos AC, Sousa MV, Barbosa NB, Mesquita LM, et al. Development of liver disease caused by chronic periodontitis in rats. J Appl Oral Sci. 2025;33:e20250550. doi: 10.1590/1678-7757-2025-0550

» https://doi.org/10.1590/1678-7757-2025-0550

7- Zhang Z, Zheng Q, Liu Y, Chen G, Li Y. Association between periodontitis and mortality in participants with metabolic dysfunction-associated steatotic liver disease: results from NHANES. BMC Oral Health. 2025;25(1):567. doi: 10.1186/s12903-025-05959-7

» https://doi.org/10.1186/s12903-025-05959-7

8- Leow WQ, Chan AW, Mendonza PG, Lo K, Yap K, Kim H. Non-alcoholic fatty liver disease: the pathologist's perspective. Clin Mol Hepatol. 2023;29(Suppl):S302-S318. doi: 10.3350/cmh.2022.0329

» https://doi.org/10.3350/cmh.2022.0329

9- Mitra S, De A, Chowdhury A. Epidemiology of non-alcoholic and alcoholic fatty liver diseases. Transl Gastroenterol Hepatol. 2020;5:16. doi: 10.21037/tgh.2019.09.08

» https://doi.org/10.21037/tgh.2019.09.08

10- Pischke S, Shiprov A, Peters U, Schulze Zur Wiesch J, Kluwe J, Westphal T, et al. High prevalence of periodontal disease in patients with NASH-possible association of poor dental health with NASH severity. Ann Hepatol. 2023;28(2):100887. doi: 10.1016/j.aohep.2022.100887

» https://doi.org/10.1016/j.aohep.2022.100887

11- Alves EH, Carvalos AS, Silva FR, França LF, Lenardo DD, Vasconcelos AC, et al. Bromelain reduces the non-alcoholic fatty liver disease and periodontal damages caused by ligature-induced periodontitis. Oral Dis. 2020;26(8):1793-802. doi: 10.1111/odi.13476

» https://doi.org/10.1111/odi.13476

12- Andrade RS, França LF, Pessoa LS, Landim BA, Rodrigues AA, Alves EH, et al. High-fat diet aggravates the liver disease caused by periodontitis in rats. J Periodontol. 2019;90(9):1023-31. doi: 10.1002/JPER.18-0564

» https://doi.org/10.1002/JPER.18-0564

13- Pessoa LS, Silva FR, Alves EH, França LF, Lenardo DD, Carvalho JS, et al. One or two ligatures inducing periodontitis are sufficient to cause fatty liver. Med Oral Patol Oral Cir Bucal. 2018;23(3):e269-e276. doi: 10.4317/medoral.22204

» https://doi.org/10.4317/medoral.22204

14- Vasconcelos AC, Vasconcelos DF, Silva FR, França LF, Alves EH, Lenardo DD, et al. Periodontitis causes abnormalities in the liver of rats. J Periodontol. 2019;90(3):295-305. doi: 10.1002/JPER.18-0226

» https://doi.org/10.1002/JPER.18-0226

15- Ferreira IL, Costa S, Moraes BJ, Costa A, Fokt O, Marinho D, et al. Mitochondrial and redox changes in periodontitis and type 2 diabetes human blood mononuclear cells. Antioxidants (Basel). 2023;12(2):226. doi: 10.3390/antiox12020226

» https://doi.org/10.3390/antiox12020226

16- Li Q, Ma H, Shang Y, Xin X, Liu X, Wu Z, et al. The role of uncoupling protein 2 in experimental periodontitis-associated renal injury in rats. Hua Xi Kou Qiang Yi Xue Za Zhi. 2024;42(4):502-11. doi: 10.7518/hxkq.2024.2023378

» https://doi.org/10.7518/hxkq.2024.2023378

17- Dara L. The receptor interacting protein kinases in the liver. Semin Liver Dis. 2018;38(1):73-86. doi: 10.1055/s-0038-1629924

» https://doi.org/10.1055/s-0038-1629924

18- Gao H, Zhong Y, Zhou L, Lin S, Hou X, Ding Z, et al. Kindlin-2 inhibits TNF/NF-?B-Caspase 8 pathway in hepatocytes to maintain liver development and function. Elife. 2023;12:e81792. doi: 10.7554/eLife.81792

» https://doi.org/10.7554/eLife.81792

19- Aral K, Aral CA, Kapila Y. The role of caspase-8, caspase-9, and apoptosis inducing factor in periodontal disease. J Periodontol. 2019;90(3):288-94. doi: 10.1002/JPER.17-0716

» https://doi.org/10.1002/JPER.17-0716

20- Caetano VS, Andrade RS, França LF, Pessoa LS, Rodrigues AA, Alves EH, et al. Food restriction reduces hepatic alterations associated with experimental periodontitis. J Periodontol. 2022;93(1):156-65. doi: 10.1002/JPER.20-0772

» https://doi.org/10.1002/JPER.20-0772

21- Vasconcelos AC, Vasconcelos DF, Silva FR, França LF, Alves EH, Lenardo DD, et al. Alpha-terpineol complexed with beta-cyclodextrin reduces damages caused by periodontitis in rats. J Periodontal Res. 2020;55(6):877-86. doi: 10.1111/jre.12780

» https://doi.org/10.1111/jre.12780

22- Vasconcelos DF, Silva FR, Pinto ME, Santana LA, Souza IG, Souza LK, et al. Decrease of pericytes is associated with liver disease caused by ligature-induced periodontitis in rats. J Periodontol. 2017;88(2):e49-e57. doi: 10.1902/jop.2016.160392

» https://doi.org/10.1902/jop.2016.160392

23- Molon RS, Mascarenhas VI, Avila ED, Finoti LS, Toffoli GB, Spolidorio DM, et al. Long-term evaluation of oral gavage with periodontopathogens or ligature induction of experimental periodontal disease in mice. Clin Oral Investig. 2016;20(6):1203-16. doi: 10.1007/s00784-015-1607-0

» https://doi.org/10.1007/s00784-015-1607-0

24- Lee MJ, Ryu HH, Hwang JW, Kim JR, Cho ES, Choi JK, et al. Sirt6 activation ameliorates inflammatory bone loss in ligature-induced periodontitis in mice. Int J Mol Sci. 2023;24(13):10714. doi: 10.3390/ijms241310714

» https://doi.org/10.3390/ijms241310714

25- Lin P, Niimi H, Ohsugi Y, Tsuchiya Y, Shimohira T, Komatsu K, et al. Application of ligature-induced periodontitis in mice to explore the molecular mechanism of periodontal disease. Int J Mol Sci. 2021;22(16):8900. doi: 10.3390/ijms22168900

» https://doi.org/10.3390/ijms22168900

26- Ambati M, Rani KR, Reddy PV, Suryprasanna J, Dasari R, Gireddy H. Evaluation of oxidative stress in chronic periodontitis patients following systemic antioxidant supplementation: a clinical and biochemical study. J Nat Sci Biol Med. 2017;8(1):99-103. doi: 10.4103/0976-9668.198366

» https://doi.org/10.4103/0976-9668.198366

27- Mirzaei A, Shahrestanaki E, Malmir H, Ejtahed HS, Tajbakhsh D, Seif E, et al. Association of periodontitis with lipid profile: an updated systematic review and meta-analysis. J Diabetes Metab Disord. 2022;21(2):1377-93. doi: 10.1007/s40200-022-01071-7

» https://doi.org/10.1007/s40200-022-01071-7

28- Newton K, Wickliffe KE, Maltzman A, Dugger DL, Reja R, Zhang Y, et al. Activity of caspase-8 determines plasticity between cell death pathways. Nature. 2019;575(7784):679-82. doi: 10.1038/s41586-019-1752-8

» https://doi.org/10.1038/s41586-019-1752-8

29- Pradeep AR, Suke DK, Prasad MV, Singh SP, Martande SS, Nagpal K, et al. Expression of key executioner of apoptosis caspase-3 in periodontal health and disease. J Investig Clin Dent. 2016;7(2):174-9. doi: 10.1111/jicd.12134

» https://doi.org/10.1111/jicd.12134

30- Li CP, Li JH, He SY, Li P, Zhong XL. Roles of Fas/Fasl, Bcl-2/Bax, and Caspase-8 in rat nonalcoholic fatty liver disease pathogenesis. Genet Mol Res. 2014;13(2):3991-9. doi: 10.4238/2014.May.23.10

» https://doi.org/10.4238/2014.May.23.10

31- Zhang Y, Zhou G, Chen Z, Guan W, Zhang J, Bi M, et al. Si-Wu-Tang alleviates nonalcoholic fatty liver disease via blocking TLR4-JNK and Caspase-8-GSDMD signaling pathways. Evid Based Complement Alternat Med. 2020;2020:8786424. doi: 10.1155/2020/8786424

» https://doi.org/10.1155/2020/8786424

32- Tummers B, Green DR. Caspase-8: regulating life and death. Immunol Rev. 2017;277(1):76-89. doi: 10.1111/imr.12541

» https://doi.org/10.1111/imr.12541

33- Prasun P, Ginevic I, Oishi K. Mitochondrial dysfunction in nonalcoholic fatty liver disease and alcohol related liver disease. Transl Gastroenterol Hepatol. 2021;6:4. doi: 10.21037/tgh-20-125

» https://doi.org/10.21037/tgh-20-125

34- Alikhani M, Alikhani Z, Raptis M, Graves DT. TNF-alpha in vivo stimulates apoptosis in fibroblasts through caspase-8 activation and modulates the expression of pro-apoptotic genes. J Cell Physiol. 2004;201(3):341-8. doi: 10.1002/jcp.20067

» https://doi.org/10.1002/jcp.20067

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Published

2026-04-06

Issue

Vol. 34 (2026)

Section

Original Articles

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How to Cite

Silva, F. R. P. da, Martins, V. B. D., Nascimento, H. M. S., Gomes, P. R. C., Barbosa, N. B., Caetano, V. da S., Diniz, I. M. A., Silva, K. D. da, Vasconcelos, A. C. C. G., & Vasconcelos, D. F. P. (2026). Expression of caspase-8 in non-alcoholic fatty liver disease related to ligature-induced periodontitis in rats. Journal of Applied Oral Science, 34, e20250717. https://doi.org/10.1590/1678-7765-2025-0717