Involvement of gap junctions in propylthiouracil‑induced cytotoxicity in BRL‑3A cells

Tang,Nan; Cai,Ziqing; Chen,Hongpeng; Cao,Longbin; Chen,Bo; Lin,Bihua

doi:10.3892/etm.2019.7244

Involvement of gap junctions in propylthiouracil‑induced cytotoxicity in BRL‑3A cells

Authors:
- Nan Tang
- Ziqing Cai
- Hongpeng Chen
- Longbin Cao
- Bo Chen
- Bihua Lin
View Affiliations

Affiliations: School of Pharmacy, Guangdong Medical University, Dongguan, Guangdong 523808, P.R. China, School of Information Engineering, Guangdong Medical University, Dongguan, Guangdong 523808, P.R. China, School of Basic Medical Sciences, Guangdong Medical University, Dongguan, Guangdong 523808, P.R. China
Published online on: February 6, 2019 https://doi.org/10.3892/etm.2019.7244
Pages: 2799-2806

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Gap junctions (GJs), which are important plasma membrane channels for the transfer of signaling molecules between adjacent cells, have been implicated in drug‑induced liver injury. However, the influence and the underlying mechanisms of GJs in propylthiouracil (PTU)‑induced hepatotoxicity are unclear. In the present study, distinct manipulations were performed to regulate GJ function in the BRL‑3A rat liver cell line. The results indicated that the toxic effect of PTU in BRL‑3A cells was mediated by GJ intercellular communication, as cell death was significantly attenuated in the absence of functional GJ channels. Furthermore, the specific knockdown of connexin‑32 (Cx32; a major GJ component protein in hepatocytes) using small interfering RNA was observed to decrease necrosis, intracellular PTU content and the level of reactive oxygen species (ROS) following PTU exposure. These observations demonstrated that suppressing GJ Cx32 could confer protection against PTU‑induced cytotoxicity through decreasing the accumulation of PTU and ROS. To the best of our knowledge, the present study is the first to demonstrate the role and possible underlying mechanisms of GJs in the regulation of PTU‑induced toxicity in BRL‑3A rat liver cells.

View Figures

View References

1	Landau RL: Landmark perspective: Treatment of hyperthyroidism. JAMA. 251:1747–1748. 1984. View Article : Google Scholar : PubMed/NCBI
2	Glinoer D and Cooper DS: The propylthiouracil dilemma. Curr Opin Endocrinol Diabetes Obes. 19:402–407. 2012. View Article : Google Scholar : PubMed/NCBI
3	Malozowski S and Chiesa A: Propylthiouracil-induced hepatotoxicity and death. Hopefully, never more. J Clin Endocrinol Metab. 95:3161–3163. 2010. View Article : Google Scholar : PubMed/NCBI
4	Yang J, Li LF, Xu Q, Zhang J, Weng WW, Zhu YJ and Dong MJ: Analysis of 90 cases of antithyroid drug-induced severe hepatotoxicity over 13 years in China. Thyroid. 25:278–283. 2015. View Article : Google Scholar : PubMed/NCBI
5	Wang MT, Lee WJ, Huang TY, Chu CL and Hsieh CH: Antithyroid drug-related hepatotoxicity in hyperthyroidism patients: A population-based cohort study. Br J Clin Pharmacol. 78:619–629. 2014. View Article : Google Scholar : PubMed/NCBI
6	Wu DB, Chen EQ, Bai L and Tang H: Propylthiouracil-induced liver failure and artificial liver support systems: A case report and review of the literature. Ther Clin Risk Manag. 13:65–68. 2017. View Article : Google Scholar : PubMed/NCBI
7	Bahn RS, Burch HS, Cooper DS, Garber JR, Greenlee CM, Klein IL, Laurberg P, McDougall IR, Rivkees SA, Ross D, et al: The role of propylthiouracil in the management of graves' disease in adults: Report of a meeting jointly sponsored by the american thyroid association and the food and drug administration. Thyroid. 19:673–674. 2009. View Article : Google Scholar : PubMed/NCBI
8	Williams KV, Nayak S, Becker D, Reyes J and Burmeister LA: Fifty years of experience with propylthiouracil-associated hepatotoxicity: What have we learned? J Clin Endocrinol Metab. 82:1727–1733. 1997. View Article : Google Scholar : PubMed/NCBI
9	Maeda S and Tsukihara T: Structure of the gap junction channel and its implications for its biological functions. Cell Mol Life Sci. 68:1115–1129. 2011. View Article : Google Scholar : PubMed/NCBI
10	Sosinsky GE and Nicholson BJ: Structural organization of gap junction channels. Biochim Biophys Acta. 1711:99–125. 2005. View Article : Google Scholar : PubMed/NCBI
11	Harris AL: Emerging issues of connexin channels: Biophysics fills the gap. Q Rev Biophys. 34:325–472. 2001. View Article : Google Scholar : PubMed/NCBI
12	Koffler LD, Fernstrom MJ, Akiyama TE, Gonzalez FJ and Ruch RJ: Positive regulation of connexin32 transcription by hepatocyte nuclear factor-1alpha. Arch Biochem Biophys. 407:160–167. 2002. View Article : Google Scholar : PubMed/NCBI
13	Zoidl G and Dermietzel R: Gap junctions in inherited human disease. Pflugers Arch. 460:451–466. 2010. View Article : Google Scholar : PubMed/NCBI
14	Scott DA, Kraft ML, Stone EM, Sheffield VC and Smith RJ: Connexin mutations and hearing loss. Nature. 391:321998. View Article : Google Scholar : PubMed/NCBI
15	Belousov AB, Fontes JD, Freitas-Andrade M and Naus CC: Gap junctions and hemichannels: Communicating cell death in neurodevelopment and disease. BMC Cell Biol. 18 (Suppl 1):42017. View Article : Google Scholar : PubMed/NCBI
16	Park WJ, Park JW, Erez-Roman R, Kogot-Levin A, Bame JR, Tirosh B, Saada A, Merrill AH Jr, Pewzner-Jung Y and Futerman AH: Protection of a ceramide synthase 2 null mouse from drug-induced liver injury: Role of gap junction dysfunction and connexin 32 mislocalization. J Biol Chem. 288:30904–30916. 2013. View Article : Google Scholar : PubMed/NCBI
17	Maurel M and Rosenbaum J: Closing the gap on drug-induced liver injury. Hepatology. 56:781–783. 2012. View Article : Google Scholar : PubMed/NCBI
18	Naiki-Ito A, Asamoto M, Naiki T, Ogawa K, Takahashi S, Sato S and Shirai T: Gap junction dysfunction reduces acetaminophen hepatotoxicity with impact on apoptotic signaling and connexin 43 protein induction in rat. Toxicol Pathol. 38:280–286. 2010. View Article : Google Scholar : PubMed/NCBI
19	Patel SJ, Milwid JM, King KR, Bohr S, Iracheta-Vellve A, Li M, Vitalo A, Parekkadan B, Jindal R and Yarmush ML: Gap junction inhibition prevents drug-induced liver toxicity and fulminant hepatic failure. Nat Biotechnol. 30:179–183. 2012. View Article : Google Scholar : PubMed/NCBI
20	Asamoto M, Hokaiwado N, Murasaki T and Shirai T: Connexin 32 dominant-negative mutant transgenic rats are resistant to hepatic damage by chemicals. Hepatology. 40:205–210. 2004. View Article : Google Scholar : PubMed/NCBI
21	Huang F, Li S, Gan X, Wang R and Chen Z: Propofol inhibits gap junctions by attenuating sevoflurane-induced cytotoxicity against rat liver cells in vitro. Eur J Anaesthesiol. 31:219–224. 2014. View Article : Google Scholar : PubMed/NCBI
22	Wang Q, You T, Yuan D, Han X, Hong X, He B, Wang L, Tong X, Tao L and Harris AL: Cisplatin and oxaliplatin inhibit gap junctional communication by direct action and by reduction of connexin expression, thereby counteracting cytotoxic efficacy. J Pharmacol Exp Ther. 333:903–911. 2010. View Article : Google Scholar : PubMed/NCBI
23	Tang N, Liu J, Chen B, Zhang Y, Yu M, Cai Z and Chen H: Effects of gap junction intercellular communication on the docetaxel-induced cytotoxicity in rat hepatocytes. Mol Med Rep. 15:2689–2694. 2017. View Article : Google Scholar : PubMed/NCBI
24	Wang R, Huang F, Chen Z and Li S: Downregulation of connexin 32 attenuates hypoxia/reoxygenation injury in liver cells. J Biochem Mol Toxicol. 29:189–197. 2015. View Article : Google Scholar : PubMed/NCBI
25	Hong X, Wang Q, Yang Y, Zheng S, Tong X, Zhang S, Tao L and Harris AL: Gap junctions propagate opposite effects in normal and tumor testicular cells in response to cisplatin. Cancer Lett. 317:165–171. 2012. View Article : Google Scholar : PubMed/NCBI
26	Zhang Y, Tao L, Fan L, Peng Y, Yang K, Zhao Y, Song Q and Wang Q: Different gap junction-propagated effects on cisplatin transfer result in opposite responses to cisplatin in normal cells versus tumor cells. Sci Rep. 5:125632015. View Article : Google Scholar : PubMed/NCBI
27	Tang N, Chen H, Cai Z, Zhan P, Pan T, Zhang Y and Wu X: Development and validation of RP-HPLC method for determination of propylthiouracil in hepatic cells. Curr Pharm Anal. 14:219–222. 2018. View Article : Google Scholar
28	Tian YY, An LJ, Jiang L, Duan YL, Chen J and Jiang B: Catalpol protects dopaminergic neurons from LPS-induced neurotoxicity in mesencephalic neuron-glia cultures. Life Sci. 80:193–199. 2006. View Article : Google Scholar : PubMed/NCBI
29	Davidson JS and Baumgarten IM: Glycyrrhetinic acid derivatives: A novel class of inhibitors of gap-junctional intercellular communication. Structure-activity relationships. J Pharmacol Exp Ther. 246:1104–1107. 1988.PubMed/NCBI
30	Vrionis FD, Wu JK, Qi P, Waltzman M, Cherington V and Spray DC: The bystander effect exerted by tumor cells expressing the herpes simplex virus thymidine kinase (HSVtk) gene is dependent on connexin expression and cell communication via gap junctions. Gene Ther. 4:577–585. 1997. View Article : Google Scholar : PubMed/NCBI
31	Hamada N, Matsumoto H, Hara T and Kobayashi Y: Intercellular and intracellular signaling pathways mediating ionizing radiation-induced bystander effects. J Radiat Res. 48:87–95. 2007. View Article : Google Scholar : PubMed/NCBI
32	Prise KM and O'Sullivan JM: Radiation-induced bystander signalling in cancer therapy. Nat Rev Cancer. 9:351–360. 2009. View Article : Google Scholar : PubMed/NCBI
33	Krutovskikh VA, Piccoli C and Yamasaki H: Gap junction intercellular communication propagates cell death in cancerous cells. Oncogene. 21:1989–1999. 2002. View Article : Google Scholar : PubMed/NCBI
34	Krysko DV, Leybaert L, Vandenabeele P and D'Herde K: Gap junctions and the propagation of cell survival and cell death signals. Apoptosis. 10:459–469. 2005. View Article : Google Scholar : PubMed/NCBI
35	Lin JH, Weigel H, Cotrina ML, Liu S, Bueno E, Hansen AJ, Hansen TW, Goldman S and Nedergaard M: Gap-junction-mediated propagation and amplification of cell injury. Nat Neurosci. 1:7431998. View Article : Google Scholar
36	Decrock E, Krysko DV, Vinken M, Kaczmarek A, Crispino G, Bol M, Wang N, De Bock M, De Vuyst E, Naus CC, et al: Transfer of IP3 through gap junctions is critical, but not sufficient, for the spread of apoptosis. Cell Death Differ. 19:947–957. 2012. View Article : Google Scholar : PubMed/NCBI
37	Decrock E, Vinken M, Bol M, D'Herde K, Rogiers V, Vandenabeele P, Krysko DV, Bultynck G and Leybaert L: Calcium and connexin-based intercellular communication, a deadly catch? Cell Calcium. 50:310–321. 2011. View Article : Google Scholar : PubMed/NCBI
38	Niessen H, Harz H, Bedner P, Krämer K and Willecke K: Selective permeability of different connexin channels to the second messenger inositol 1,4,5-trisphosphate. J Cell Sci. 113:1365–1372. 2000.PubMed/NCBI
39	Goldberg GS, Moreno AP and Lampe PD: Gap junctions between cells expressing connexin 43 or 32 show inverse permselectivity to adenosine and ATP. J Biol Chem. 277:36725–36730. 2002. View Article : Google Scholar : PubMed/NCBI
40	Bevans CG, Kordel M, Rhee SK and Harris AL: Isoform composition of connexin channels determines selectivity among second messengers and uncharged molecules. J Biol Chem. 273:2808–2816. 1998. View Article : Google Scholar : PubMed/NCBI
41	Kariya K, Sawahata T, Okuno S and Lee E: Inhibition of hepatic glutathione transferases by propylthiouracil and its metabolites. Biochem Pharmacol. 35:1475–1479. 1986. View Article : Google Scholar : PubMed/NCBI
42	Ye J, Zhong X, Du Y, Cai C and Pan T: Role of levothyroxine and vitamin E supplementation in the treatment of oxidative stress-induced injury and apoptosis of myocardial cells in hypothyroid rats. J Endocrinol Invest. 40:713–719. 2017. View Article : Google Scholar : PubMed/NCBI
43	Tas S, Dirican M, Sarandöl E and Serdar Z: The effect of taurine supplementation on oxidative stress in experimental hypothyroidism. Cell Biochem Funct. 24:153–158. 2006. View Article : Google Scholar : PubMed/NCBI
44	Schattenberg JM, Wang Y, Rigoli RM, Koop DR and Czaja MJ: CYP2E1 overexpression alters hepatocyte death from menadione and fatty acids by activation of ERK1/2 signaling. Hepatology. 39:444–455. 2004. View Article : Google Scholar : PubMed/NCBI
45	Zheng L, Wang C, Luo T, Lu B, Ma H, Zhou Z, Zhu D, Chi G, Ge P and Luo Y: JNK activation contributes to oxidative stress-induced parthanatos in glioma cells via increase of intracellular ROS production. Mol Neurobiol. 54:3492–3505. 2017. View Article : Google Scholar : PubMed/NCBI
46	Muriel P: Role of free radicals in liver diseases. Hepatol Int. 3:526–536. 2009. View Article : Google Scholar : PubMed/NCBI
47	Luo C, Yuan D, Li X, Yao W, Luo G, Chi X, Li H, Irwin MG, Xia Z and Hei Z: Propofol attenuated acute kidney injury after orthotopic liver transplantation via inhibiting gap junction composed of connexin 32. Anesthesiology. 122:72–86. 2015. View Article : Google Scholar : PubMed/NCBI