Heme Oxygenase-1 Attenuates Cadmium-induced Mitochondrial-caspase 3- Dependent Apoptosis in Human Hepatoma Cell Line

Overview

Journal BMC Pharmacol Toxicol

Publisher Biomed Central

Specialty Pharmacology

Date 2015 Dec 17

PMID 26670903

Citations 14

Authors

Akeem O Lawal

Jeanine L Marnewick

Elizabeth M Ellis

Affiliations

Soon will be listed here.

Abstract

Background: Cadmium (Cd) is a well known environmental and industrial toxicant causing damaging effects in numerous organs. In this study, we examined the role of heme oxygenase-1 (HO-1) in modulating the Cd-induced apoptosis in human hepatoma (HepG2) cells after 24 h exposure.

Methods: HepG2 cells were exposed to 5 and 10 μM Cd as CdCl2 for 24 h while other sets of cells were pre-treated with either 10 μM Cobalt protoporphyrin (CoPPIX) or 10 μM Tin protoporphyrin (SnPPIX) for 24 h, or 50 μM Z-DEVD-FMK for 1 h before exposure to 5 and 10 μM CdCl2 for 24 h. Expressions of caspase 3, cytosolic cytochrome c, mitochondrial Bax and anti-apoptotic BCL-xl proteins were assessed by western blot. Intracellular reactive oxygen species (ROS) production was determined using the dihydrofluorescein diacetate (H2DFA) method. Cell viability was assessed by MTT assay, while a flow cytometry method was used to assess the level of apoptosis in the cell populations.

Results: Our results show that there were a significant increase in the expression of cytosolic cytochrome c, mitochondrial Bax protein, and caspase 3 at 5 and 10 μM compared to the control, but these increases were attenuated by the presence of CoPPIX. The presence of SnPPIX significantly enhanced Cd-induced caspase 3 activities. CoPPIX significantly decreased the level of ROS production by 24.6 and 22.2 % in 5 and 10 μM CdCl2, respectively, but SnPPIX caused a significant increase in ROS production in the presence of CdCl2. HepG2 cell viability was also significantly impaired by 13.89 and 32.53 % in the presence of 5 and 10 μM CdCl2, respectively, but the presence of CoPPIX and Z-DEVD-FMK significantly enhanced cell survival, while SnPPIX enhanced Cd-impaired cell viability. The presence of CoPPIX and Z-DEVD-FMK also significantly decreased the population of apoptotic and necrotic cells compared with Cd.

Conclusion: In summary, the present study showed that HO-1 attenuates the Cd-induced caspase 3 dependent pathway of apoptosis in HepG2 cells, probably by modulating Cd-induced oxidative stress.

Citing Articles

Pretreatment with aqueous Lam. leaf extract prevents cadmium-induced hepatotoxicity by improving cellular antioxidant machinery and reducing cadmium accumulation.

Buranasudja V, Sanookpan K, Vimolmangkang S, Binalee A, Mika K, Krobthong S Heliyon. 2024; 10(18):e37424.

PMID: 39309955 PMC: 11416483. DOI: 10.1016/j.heliyon.2024.e37424.

Placental BCRP transporter reduces cadmium accumulation and toxicity in immortalized human trophoblasts.

Zhang R, Walker L, Wen X, Doherty C, Gorczyca L, Buckley B Reprod Toxicol. 2023; 121:108466.

PMID: 37660740 PMC: 10591833. DOI: 10.1016/j.reprotox.2023.108466.

Potential effect of luteolin, epiafzelechin, and albigenin on rats under cadmium-induced inflammatory insult: and approach.

Shahzadi A, Tariq N, Sonmez H, Waquar S, Zahid A, Javed M Front Chem. 2023; 11:1036478.

PMID: 36936530 PMC: 10016615. DOI: 10.3389/fchem.2023.1036478.

Oxidant- induced preconditioning: A pharmacologic approach for triggering renal 'self defense'.

Zager R Physiol Rep. 2022; 10(20):e15507.

PMID: 36305701 PMC: 9615572. DOI: 10.14814/phy2.15507.

Neuroprotective Effects of the Inert Gas Argon on Experimental Traumatic Brain Injury In Vivo with the Controlled Cortical Impact Model in Mice.

Schneider F, Krieg S, Lindauer U, Stoffel M, Ryang Y Biology (Basel). 2022; 11(2).

PMID: 35205025 PMC: 8869506. DOI: 10.3390/biology11020158.

References

Lee W, Thevenod F . Novel roles for ceramides, calpains and caspases in kidney proximal tubule cell apoptosis: lessons from in vitro cadmium toxicity studies. Biochem Pharmacol. 2008; 76(11):1323-32. DOI: 10.1016/j.bcp.2008.07.004. View

Pae H, Lee Y, Chung H . Heme oxygenase-1 and carbon monoxide: emerging therapeutic targets in inflammation and allergy. Recent Pat Inflamm Allergy Drug Discov. 2008; 2(3):159-65. DOI: 10.2174/187221308786241929. View

Tang C, Lu Y, Xie J, Wang F, Zou J, Yang J . Downregulation of survivin and activation of caspase-3 through the PI3K/Akt pathway in ursolic acid-induced HepG2 cell apoptosis. Anticancer Drugs. 2009; 20(4):249-58. DOI: 10.1097/CAD.0b013e328327d476. View

Hempel S, Buettner G, OMalley Y, Wessels D, Flaherty D . Dihydrofluorescein diacetate is superior for detecting intracellular oxidants: comparison with 2',7'-dichlorodihydrofluorescein diacetate, 5(and 6)-carboxy-2',7'-dichlorodihydrofluorescein diacetate, and dihydrorhodamine 123. Free Radic Biol Med. 1999; 27(1-2):146-59. DOI: 10.1016/s0891-5849(99)00061-1. View

Qu W, Diwan B, Reece J, Bortner C, Pi J, Liu J . Cadmium-induced malignant transformation in rat liver cells: role of aberrant oncogene expression and minimal role of oxidative stress. Int J Cancer. 2004; 114(3):346-55. DOI: 10.1002/ijc.20736. View

Abraham N, Kappas A . Heme oxygenase and the cardiovascular-renal system. Free Radic Biol Med. 2005; 39(1):1-25. DOI: 10.1016/j.freeradbiomed.2005.03.010. View

Oh S, Lim S . A rapid and transient ROS generation by cadmium triggers apoptosis via caspase-dependent pathway in HepG2 cells and this is inhibited through N-acetylcysteine-mediated catalase upregulation. Toxicol Appl Pharmacol. 2005; 212(3):212-23. DOI: 10.1016/j.taap.2005.07.018. View

Tsoyi K, Lee T, Lee Y, Kim H, Seo H, Lee J . Heme-oxygenase-1 induction and carbon monoxide-releasing molecule inhibit lipopolysaccharide (LPS)-induced high-mobility group box 1 release in vitro and improve survival of mice in LPS- and cecal ligation and puncture-induced sepsis model in vivo. Mol Pharmacol. 2009; 76(1):173-82. DOI: 10.1124/mol.109.055137. View

Liu J, Qu W, Kadiiska M . Role of oxidative stress in cadmium toxicity and carcinogenesis. Toxicol Appl Pharmacol. 2009; 238(3):209-14. PMC: 4287357. DOI: 10.1016/j.taap.2009.01.029. View

10.

Hossain S, Liu H, Nguyen M, Shore G, Almazan G . Cadmium exposure induces mitochondria-dependent apoptosis in oligodendrocytes. Neurotoxicology. 2009; 30(4):544-54. DOI: 10.1016/j.neuro.2009.06.001. View

11.

Ji C, Ren F, Ma H, Xu M . The roles of p38MAPK and caspase-3 in DADS-induced apoptosis in human HepG2 cells. J Exp Clin Cancer Res. 2010; 29:50. PMC: 2890544. DOI: 10.1186/1756-9966-29-50. View

12.

Lawal A, Ellis E . Differential sensitivity and responsiveness of three human cell lines HepG2, 1321N1 and HEK 293 to cadmium. J Toxicol Sci. 2010; 35(4):465-78. DOI: 10.2131/jts.35.465. View

13.

Lawal A, Ellis E . Nrf2-mediated adaptive response to cadmium-induced toxicity involves protein kinase C delta in human 1321N1 astrocytoma cells. Environ Toxicol Pharmacol. 2011; 32(1):54-62. DOI: 10.1016/j.etap.2011.03.010. View

14.

Lawal A, Ellis E . The chemopreventive effects of aged garlic extract against cadmium-induced toxicity. Environ Toxicol Pharmacol. 2011; 32(2):266-74. DOI: 10.1016/j.etap.2011.05.012. View

15.

Yang Y, Lii C, Lin A, Yeh Y, Yao H, Li C . Induction of glutathione synthesis and heme oxygenase 1 by the flavonoids butein and phloretin is mediated through the ERK/Nrf2 pathway and protects against oxidative stress. Free Radic Biol Med. 2011; 51(11):2073-81. DOI: 10.1016/j.freeradbiomed.2011.09.007. View

16.

Lawal A, Ellis E . Phospholipase C mediates cadmium-dependent apoptosis in HEK 293 cells. Basic Clin Pharmacol Toxicol. 2011; 110(6):510-7. DOI: 10.1111/j.1742-7843.2011.00843.x. View

17.

Nguyen K, Willmore W, Tayabali A . Cadmium telluride quantum dots cause oxidative stress leading to extrinsic and intrinsic apoptosis in hepatocellular carcinoma HepG2 cells. Toxicology. 2013; 306:114-23. DOI: 10.1016/j.tox.2013.02.010. View

18.

Yuan C, Kadiiska M, Achanzar W, Mason R, Waalkes M . Possible role of caspase-3 inhibition in cadmium-induced blockage of apoptosis. Toxicol Appl Pharmacol. 2000; 164(3):321-9. DOI: 10.1006/taap.2000.8921. View

19.

He X, Chen M, Ma Q . Activation of Nrf2 in defense against cadmium-induced oxidative stress. Chem Res Toxicol. 2008; 21(7):1375-83. DOI: 10.1021/tx800019a. View

20.

Belyaeva E, Dymkowska D, Wieckowski M, Wojtczak L . Mitochondria as an important target in heavy metal toxicity in rat hepatoma AS-30D cells. Toxicol Appl Pharmacol. 2008; 231(1):34-42. DOI: 10.1016/j.taap.2008.03.017. View