Co-Administration Exposure Toxicity of Isoniazid and Rifampicin in Chang Liver Cell Lines
Abstract
The study is designed to investigate the exposure toxicity of isoniazid (INH) and rifampicin (RIF) in combination on Chang liver cell lines. Each 50, 100, and 200 ng/ml of INH and RIF were co-administered to the Chang cell lines for a period of 24 hours; the untreated cell lines were served as a control. Alanine aminotransferase (ALT), Aspartate aminotransferase (AST) and alkaline phosphatase (ALP) activity in the culture medium were assayed to determine the integrity of the cell lines. β-Galactosidase and cathepsin activities were also assayed in the lysosomal fraction. A comet assay was performed to determine DNA damage in the cell lines. The mRNA expressions of CYP2E1, BAX, BCL2, P53, and caspase-3 were studied. Cells treated with INH and RIF showed abnormal levels of ALP, AST, and ALT activities in the culture medium. However, no such abnormalities were inferred in the medium of the unexposed control cells. Altered enzymatic activities were seen in the lysosomal fractions of the treated cells, whereas no such abnormalities were detected in the untreated control cell lines. 71% of DNA damage was recorded in the intoxicated cells, whereas the DNA of the control cell lines remained intact. A significant increase in the CYP2E1, P53, and caspase-3, along with alterations in BAX and BCL2 mRNA expressions, was recorded in the intoxicated cells. In contrast, the untreated control cells showed optimal mRNA expression. This study demonstrated that exposure to INH and RIF in Chang liver cell lines compromised cellular integrity, altered the mRNA expression, and induced significant DNA damage.
Keywords:
Apoptosis, CYP2E1, Isoniazid, Oxidative Stress, Rifampicin
Downloads
Download data is not yet available.
References
Anson, M. L. (1938). The estimation of pepsin, trypsin, papain, and cathepsin with hemoglobin. The Journal of General Physiology, 22(1), 79-89. https://doi.org/10.1085/jgp.22.1.79
Batool, Z., Wu, Q., Weng, G., & Shen, B. (2026). A Review on Isoniazid and Rifampicin‐Associated Hepatotoxicity: From Metabolism to Immunity. Archiv der Pharmazie, 359(3), e70204. https://doi.org/10.1002/ardp.70204
Beyfuss, K., & Hood, D. A. (2018). A systematic review of p53 regulation of oxidative stress in skeletal muscle. Redox Report, 23(1), 100-117.https://doi.org/10.1080/13510002.2017.1416773
Bishnu, D., Santra, S., Purkait, S., & Santra, A. (2025). Unraveling the Role of CYP2E1 in Antitubercular Drug–Induced Hepatotoxicity: From Molecular Mechanisms to Clinical Implications. International Journal of Hepatology, 2025(1), 9980298. https://doi.org/10.1155/ijh/9980298
Chopra, S., & Griffin, P. H. (1985). Laboratory tests and diagnostic procedures in evaluation of liver disease. The American Journal of Medicine, 79(2), 221-230. https://doi.org/10.1016/0002-9343(85)90013-0
Conchie, J., &Levvy, G. A. (1957). Inhibition of glycosidases by aldonolactones of corresponding configuration. Biochemical Journal, 65(2), 389-395. https://doi.org/10.1042/bj0650389
Dou, H., Yu, P. Y., Liu, Y. Q., Zhu, Y., Li, F. C., Wang, Y. Y., ... & Xiao, M. (2024). Recent advances in caspase-3, breast cancer, and traditional Chinese medicine: A review. Journal of Chemotherapy, 36(5), 370-388.https://doi.org/10.1080/1120009X.2023.2278014
Ekwall, B., & Acosta, D. (1982). In vitro comparative toxicity of selected drugs and chemicals in HeLa cells, Chang liver cells, and rat hepatocytes. Drug and Chemical Toxicology, 5(3), 219-231. https://doi.org/10.3109/01480548209041054
González, L. T., Minsky, N. W., Espinosa, L. E. M., Aranda, R. S., Meseguer, J. P., & Pérez, P. C. (2017). In vitro assessment of hepatoprotective agents against damage induced by acetaminophen and CCl4. BMC Complementary and Alternative Medicine, 17(1), 39.https://doi.org/10.1186/s12906-016-1506-1
Gopisetty, G., Ramachandran, K., & Singal, R. (2006). DNA methylation and apoptosis. Molecular Immunology, 43(11), 1729-1740. https://doi.org/10.1016/j.molimm.2005.11.010
He, X., Song, Y., Wang, L., & Xu, J. (2020). Protective effect of pyrrolidine dithiocarbamate on isoniazid/rifampicin-induced liver injury in rats. Molecular Medicine Reports, 21(1), 463-469. https://doi.org/10.3892/mmr.2019.10817
Killingsworth, Z. K., Misare, K. R., Ryan, A. S., Ampolini, E. A., Mendenhall, T. T., Engevik, M. A., & Hartman, J. H. (2024). Subcellular expression of CYP2E1 in HepG2 cells impacts response to free oleic and palmitic acid. Current Research in Toxicology, 7, 100195. https://doi.org/10.1016/j.crtox.2024.100195
Lin, Q., Zhang, J., Qi, J., Tong, J., Chen, S., Zhang, S., ... & Niu, J. (2025). Hepatocyte‐Derived FGF1 Alleviates Isoniazid and Rifampicin‐Induced Liver Injury by Regulating HNF4α‐Mediated Bile Acids Synthesis. Advanced Science, 12(7), 2408688. https://doi.org/10.1002/advs.202408688
Liu, D., & Xu, Y. (2011). p53, oxidative stress, and aging. Antioxidants & redox signaling, 15(6), 1669-1678. https://doi.org/10.1089/ars.2010.3644
Liu, X., Zhao, M., Mi, J., Chen, H., Sheng, L., & Li, Y. (2017). Protective effect of bicyclol on anti-tuberculosis drug induced liver injury in rats. Molecules, 22(4), 524. https://doi.org/10.3390/molecules22040524
Møller, P., Gajski, G., Gerić, M., Giovannelli, L., Azqueta, A., Haveric, A., ... & Ladeira, C. (2025). The comet assay as a tool in human biomonitoring studies: Effects of confounding factors. Mutation Research-Reviews in Mutation Research, 108566.https://doi.org/10.1016/j.mrrev.2025.108566
Naji, K. M., Al-Khatib, B. Y., Al-Haj, N. S., & D’souza, M. R. (2021). Hepatoprotective activity of melittin on isoniazid-and rifampicin-induced liver injuries in male albino rats. BMC Pharmacology and Toxicology, 22(1), 39.https://doi.org/10.1101/2020.09.08.287094
Pal, R., Rana, S., Vaiphei, K., & Singh, K. (2008). Effect of different doses of carotenoids in isoniazid-rifampicin induced hepatotoxicity in rats. Tropical gastroenterology : official journal of the Digestive Diseases Foundation, 29(3), 153–159. https://scispace.com/pdf/effect-of-different-doses-of-carotenoids-in-isoniazid-33fsouenyr.pdf
Plummer, D. T. (1987). An introduction to practical biochemistry (pp. 236-237). London: McGraw-Hill Book Company.https://doi.org/10.1016/0307-4412(78)90089-4
Putri, G. F. T. &Nurfatihah, Z. (2025). Bax/Bcl-2 ratio as the golden marker of apoptosis: molecular mechanisms and regulatory pathways. International Journal of Cell and Biomedical Science, 4(10), 309-317.https://doi.org/10.59278/cbs.v4i10.65
Sankar, M., Rajkumar, J., & Sridhar, D. (2015). Hepatoprotective activity of heptoplus on isoniazid and rifampicin induced liver damage in rats. Indian Journal of Pharmaceutical Sciences, 77(5), 556-562. https://doi.org/10.4103/0250-474x.169028
Shayakhmetova, G., Bondarenko, L., & Kovalenko, V. (2012). Damage of testicular cell macromolecules and reproductive capacity of male rats following co-administration of ethambutol, rifampicin, isoniazid and pyrazinamide. Interdisciplinary Toxicology, 5(1), 9-14. https://doi.org/10.2478/v10102-012-0002-9
Shivaji, P., & Prince, S. E. (2025). Unveiling the molecular toxicity of Isoniazid and Rifampicin in tuberculosis therapy: Emerging insights and therapeutic strategies. Toxicology Mechanisms and Methods, 35(9), 1239-1270.https://doi.org/10.1080/15376516.2025.2554918
Singh, N. P., McCoy, M. T., Tice, R. R., & Schneider, E. L. (1988). A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental Cell Research, 175(1), 184-191. https://doi.org/10.1016/0014-4827(88)90265-0
Tostmann, A., Boeree, M. J., Aarnoutse, R. E., De Lange, W. C., Van Der Ven, A. J., &Dekhuijzen, R. (2008). Antituberculosis drug‐induced hepatotoxicity: concise up‐to‐date review. Journal of Gastroenterology and Hepatology, 23(2), 192-202.https://doi.org/10.1111/j.1440-1746.2007.05207.x
Wang, F., Gómez‐Sintes, R., & Boya, P. (2018). Lysosomal membrane permeabilization and cell death. Traffic, 19(12), 918-931.https://doi.org/10.1111/tra.12613
Wang, P., Pradhan, K., Zhong, X. B., & Ma, X. (2016). Isoniazid metabolism and hepatotoxicity. Acta Pharmaceutica Sinica B, 6(5), 384-392.https://doi.org/10.1016/j.apsb.2016.07.014
Zentner, I., Back, H. M., Kagan, L., Subbian, S., Nagajyothi, J., Srivastava, S., ... &Vinnard, C. (2020). Redox imbalance and oxidative DNA damage during isoniazid treatment of HIV-associated tuberculosis: A clinical and translational pharmacokinetic study. Frontiers in Pharmacology, 11, 1103. https://doi.org/10.3389/fphar.2020.01103
Zhuang, X., Li, L., Liu, T., Zhang, R., Yang, P., Wang, X., & Dai, L. (2022). Mechanisms of isoniazid and rifampicin-induced liver injury and the effects of natural medicinal ingredients: A review. Frontiers in Pharmacology, 13, 1037814. https://doi.org/10.3389/fphar.2022.1037814
Batool, Z., Wu, Q., Weng, G., & Shen, B. (2026). A Review on Isoniazid and Rifampicin‐Associated Hepatotoxicity: From Metabolism to Immunity. Archiv der Pharmazie, 359(3), e70204. https://doi.org/10.1002/ardp.70204
Beyfuss, K., & Hood, D. A. (2018). A systematic review of p53 regulation of oxidative stress in skeletal muscle. Redox Report, 23(1), 100-117.https://doi.org/10.1080/13510002.2017.1416773
Bishnu, D., Santra, S., Purkait, S., & Santra, A. (2025). Unraveling the Role of CYP2E1 in Antitubercular Drug–Induced Hepatotoxicity: From Molecular Mechanisms to Clinical Implications. International Journal of Hepatology, 2025(1), 9980298. https://doi.org/10.1155/ijh/9980298
Chopra, S., & Griffin, P. H. (1985). Laboratory tests and diagnostic procedures in evaluation of liver disease. The American Journal of Medicine, 79(2), 221-230. https://doi.org/10.1016/0002-9343(85)90013-0
Conchie, J., &Levvy, G. A. (1957). Inhibition of glycosidases by aldonolactones of corresponding configuration. Biochemical Journal, 65(2), 389-395. https://doi.org/10.1042/bj0650389
Dou, H., Yu, P. Y., Liu, Y. Q., Zhu, Y., Li, F. C., Wang, Y. Y., ... & Xiao, M. (2024). Recent advances in caspase-3, breast cancer, and traditional Chinese medicine: A review. Journal of Chemotherapy, 36(5), 370-388.https://doi.org/10.1080/1120009X.2023.2278014
Ekwall, B., & Acosta, D. (1982). In vitro comparative toxicity of selected drugs and chemicals in HeLa cells, Chang liver cells, and rat hepatocytes. Drug and Chemical Toxicology, 5(3), 219-231. https://doi.org/10.3109/01480548209041054
González, L. T., Minsky, N. W., Espinosa, L. E. M., Aranda, R. S., Meseguer, J. P., & Pérez, P. C. (2017). In vitro assessment of hepatoprotective agents against damage induced by acetaminophen and CCl4. BMC Complementary and Alternative Medicine, 17(1), 39.https://doi.org/10.1186/s12906-016-1506-1
Gopisetty, G., Ramachandran, K., & Singal, R. (2006). DNA methylation and apoptosis. Molecular Immunology, 43(11), 1729-1740. https://doi.org/10.1016/j.molimm.2005.11.010
He, X., Song, Y., Wang, L., & Xu, J. (2020). Protective effect of pyrrolidine dithiocarbamate on isoniazid/rifampicin-induced liver injury in rats. Molecular Medicine Reports, 21(1), 463-469. https://doi.org/10.3892/mmr.2019.10817
Killingsworth, Z. K., Misare, K. R., Ryan, A. S., Ampolini, E. A., Mendenhall, T. T., Engevik, M. A., & Hartman, J. H. (2024). Subcellular expression of CYP2E1 in HepG2 cells impacts response to free oleic and palmitic acid. Current Research in Toxicology, 7, 100195. https://doi.org/10.1016/j.crtox.2024.100195
Lin, Q., Zhang, J., Qi, J., Tong, J., Chen, S., Zhang, S., ... & Niu, J. (2025). Hepatocyte‐Derived FGF1 Alleviates Isoniazid and Rifampicin‐Induced Liver Injury by Regulating HNF4α‐Mediated Bile Acids Synthesis. Advanced Science, 12(7), 2408688. https://doi.org/10.1002/advs.202408688
Liu, D., & Xu, Y. (2011). p53, oxidative stress, and aging. Antioxidants & redox signaling, 15(6), 1669-1678. https://doi.org/10.1089/ars.2010.3644
Liu, X., Zhao, M., Mi, J., Chen, H., Sheng, L., & Li, Y. (2017). Protective effect of bicyclol on anti-tuberculosis drug induced liver injury in rats. Molecules, 22(4), 524. https://doi.org/10.3390/molecules22040524
Møller, P., Gajski, G., Gerić, M., Giovannelli, L., Azqueta, A., Haveric, A., ... & Ladeira, C. (2025). The comet assay as a tool in human biomonitoring studies: Effects of confounding factors. Mutation Research-Reviews in Mutation Research, 108566.https://doi.org/10.1016/j.mrrev.2025.108566
Naji, K. M., Al-Khatib, B. Y., Al-Haj, N. S., & D’souza, M. R. (2021). Hepatoprotective activity of melittin on isoniazid-and rifampicin-induced liver injuries in male albino rats. BMC Pharmacology and Toxicology, 22(1), 39.https://doi.org/10.1101/2020.09.08.287094
Pal, R., Rana, S., Vaiphei, K., & Singh, K. (2008). Effect of different doses of carotenoids in isoniazid-rifampicin induced hepatotoxicity in rats. Tropical gastroenterology : official journal of the Digestive Diseases Foundation, 29(3), 153–159. https://scispace.com/pdf/effect-of-different-doses-of-carotenoids-in-isoniazid-33fsouenyr.pdf
Plummer, D. T. (1987). An introduction to practical biochemistry (pp. 236-237). London: McGraw-Hill Book Company.https://doi.org/10.1016/0307-4412(78)90089-4
Putri, G. F. T. &Nurfatihah, Z. (2025). Bax/Bcl-2 ratio as the golden marker of apoptosis: molecular mechanisms and regulatory pathways. International Journal of Cell and Biomedical Science, 4(10), 309-317.https://doi.org/10.59278/cbs.v4i10.65
Sankar, M., Rajkumar, J., & Sridhar, D. (2015). Hepatoprotective activity of heptoplus on isoniazid and rifampicin induced liver damage in rats. Indian Journal of Pharmaceutical Sciences, 77(5), 556-562. https://doi.org/10.4103/0250-474x.169028
Shayakhmetova, G., Bondarenko, L., & Kovalenko, V. (2012). Damage of testicular cell macromolecules and reproductive capacity of male rats following co-administration of ethambutol, rifampicin, isoniazid and pyrazinamide. Interdisciplinary Toxicology, 5(1), 9-14. https://doi.org/10.2478/v10102-012-0002-9
Shivaji, P., & Prince, S. E. (2025). Unveiling the molecular toxicity of Isoniazid and Rifampicin in tuberculosis therapy: Emerging insights and therapeutic strategies. Toxicology Mechanisms and Methods, 35(9), 1239-1270.https://doi.org/10.1080/15376516.2025.2554918
Singh, N. P., McCoy, M. T., Tice, R. R., & Schneider, E. L. (1988). A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental Cell Research, 175(1), 184-191. https://doi.org/10.1016/0014-4827(88)90265-0
Tostmann, A., Boeree, M. J., Aarnoutse, R. E., De Lange, W. C., Van Der Ven, A. J., &Dekhuijzen, R. (2008). Antituberculosis drug‐induced hepatotoxicity: concise up‐to‐date review. Journal of Gastroenterology and Hepatology, 23(2), 192-202.https://doi.org/10.1111/j.1440-1746.2007.05207.x
Wang, F., Gómez‐Sintes, R., & Boya, P. (2018). Lysosomal membrane permeabilization and cell death. Traffic, 19(12), 918-931.https://doi.org/10.1111/tra.12613
Wang, P., Pradhan, K., Zhong, X. B., & Ma, X. (2016). Isoniazid metabolism and hepatotoxicity. Acta Pharmaceutica Sinica B, 6(5), 384-392.https://doi.org/10.1016/j.apsb.2016.07.014
Zentner, I., Back, H. M., Kagan, L., Subbian, S., Nagajyothi, J., Srivastava, S., ... &Vinnard, C. (2020). Redox imbalance and oxidative DNA damage during isoniazid treatment of HIV-associated tuberculosis: A clinical and translational pharmacokinetic study. Frontiers in Pharmacology, 11, 1103. https://doi.org/10.3389/fphar.2020.01103
Zhuang, X., Li, L., Liu, T., Zhang, R., Yang, P., Wang, X., & Dai, L. (2022). Mechanisms of isoniazid and rifampicin-induced liver injury and the effects of natural medicinal ingredients: A review. Frontiers in Pharmacology, 13, 1037814. https://doi.org/10.3389/fphar.2022.1037814
Statistics
7 Views | 7 Downloads
How to Cite
Madhavan, S. and Dorai, S. (2026) “Co-Administration Exposure Toxicity of Isoniazid and Rifampicin in Chang Liver Cell Lines”, International Journal of Advancement in Life Sciences Research, 9(4), pp. 206-216. doi: https://doi.org/10.31632/ijalsr.2026.v09i04.013.
Section
Research Articles
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
. 