Dexamethasone protects normal human liver cells from apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand by upregulating the expression of P-glycoproteins

Zhao,Bo; Xie,Gui‑Juan; Li,Rui‑Feng; Chen,Qing; Zhang,Xu‑Qing

doi:10.3892/mmr.2015.4458

Dexamethasone protects normal human liver cells from apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand by upregulating the expression of P-glycoproteins

Authors:
- Bo Zhao
- Gui‑Juan Xie
- Rui‑Feng Li
- Qing Chen
- Xu‑Qing Zhang
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Affiliations: Department of Infectious Diseases, Southwest Hospital, Third Military Medical University, Chongqing 400038, P.R. China
Published online on: October 21, 2015 https://doi.org/10.3892/mmr.2015.4458
Pages: 8093-8100

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Abstract

Glucocorticoids are effective for the treatment of acute-on-chronic pre-liver failure, severe chronic hepatitis B and acute liver failure; however, the mechanism underlying the effects of treatment by glucocorticoids remains to be fully elucidated. The role and detailed mechanism of how glucocorticoids prevent liver disease progression can be elucidated by investigating the apoptosis of hepatocytes following glucocorticoid treatment. P‑glycoproteins (P‑gps) also confer resistance to apoptosis induced by a diverse range of stimuli. Glucocorticoids, particularly dexamethasone (DEX), upregulate the expression of P‑gp in several tissues. In the present study, the normal human L‑02 liver cell line was used, and techniques, including immunocytochemistry, western blot analysis, flow cytometry and reverse transcription‑quantitative polymerase chain reaction analysis were used for determining the expression levels of P‑gps, and for evaluating the effect of DEX pretreatment on the expression of P‑gps. DEX (1‑10 µM) was added to the cell culture media and incubated for 24‑72 h. The results revealed that DEX upregulated the mRNA and protein levels of P‑gp in a dose‑ and time‑dependent manner. Subsequently, tumor necrosis factor‑related apoptosis‑inducing ligand (TRAIL) was used for the induction of apoptosis in the cells, followed by a terminal deoxynucleotidyl transferase dUTP nick end labeling assay to assess the apoptotic stages. The results demonstrated that apoptosis in the group of cells, which were pre‑treated with DEX was significantly lower than that in the control group. Treatment with tariquidar, a P‑gp inhibitor, reduced the anti‑apoptotic effects of DEX. These results established that DEX protects normal human liver cells from TRAIL‑induced apoptosis by upregulating the expression of P-gp. These observations may be useful for elucidating the mechanism of DEX for preventing the progression of liver disease.

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1	Levy P, Marcellin P, Martinot-Peignoux M, Degott C, Nataf J and Benhamou JP: Clinical course of spontaneous reactivation of hepatitis B virus infection in patients with chronic hepatitis B. Hepatology. 12:570–574. 1990. View Article : Google Scholar : PubMed/NCBI
2	Sarin SK, Kumar A, Almeida JA, Chawla YK, Fan ST, Garg H, de Silva HJ, Hamid SS, Jalan R, Komolmit P, et al: Acute-on-chronic liver failure: Consensus recommendations of the Asian Pacific Association for the study of the liver (APASL). Hepatol Int. 3:269–282. 2009. View Article : Google Scholar : PubMed/NCBI
3	Zhang XQ, Jiang L, You JP, Liu YY, Peng J, Zhang HY, Xu BY and Mao Q: Efficacy of short-term dexamethasone therapy in acute-on-chronic pre-liver failure. Hepatol Res. 41:46–53. 2011. View Article : Google Scholar
4	Fujiwara K, Yasui S, Yonemitsu Y, Fukai K, Arai M, Imazeki F, Suzuki A, Suzuki H, Sadahiro T, Oda S and Yokosuka O: Efficacy of combination therapy of antiviral and immunosuppressive drugs for the treatment of severe acute exacerbation of chronic hepatitis B. J Gastroenterol. 43:711–719. 2008. View Article : Google Scholar
5	Naveau S, Balian A, Capron F, Raynard B, Fallik D, Agostini H, Grangeot-Keros L, Portier A, Galanaud P, Chaput JC and Emilie D: Balance between pro and anti-inflammatory cytokines in patients with acute alcoholic hepatitis. Gastroenterol Clin Biol. 29:269–274. 2005. View Article : Google Scholar : PubMed/NCBI
6	Wang M, Shen F, Shi LH, Xi T, Li XF, Chen X and Wu MC: Protective effect of prednisolone on ischemia-induced liver injury in rats. World J Gastroenterol. 14:4332–4337. 2008. View Article : Google Scholar : PubMed/NCBI
7	Thoresen GH, Gjone IH, Gladhaug IP, Refsnes M, Ostby E and Christoffersen T: Studies of glucocorticoid enhancement of the capacity of hepatocytes to accumulate cyclic AMP. Pharmacol Toxicol. 65:175–180. 1989. View Article : Google Scholar : PubMed/NCBI
8	Jaeschke H, Gujral JS and Bajt ML: Apoptosis and necrosis in liver disease. Liver Int. 24:85–89. 2004. View Article : Google Scholar : PubMed/NCBI
9	Nieuwenhuis B, Lüth A and Kleuser B: Dexamethasone protects human fibroblasts from apoptosis via an S1P3-receptor subtype dependent activation of PKB/Akt and Bcl XL. Pharmacol Res. 61:449–459. 2010. View Article : Google Scholar
10	Haake SM, Dinh CT, Chen S, Eshraghi AA and Van De Water TR: Dexamethasone protects auditory hair cells against TNFalpha-initiated apoptosis via activation of PI3K/Akt and NFkappaB signaling. Hear Res. 255:22–32. 2009. View Article : Google Scholar : PubMed/NCBI
11	Li M, Chen F, Liu CP, Li DM, Li X, Wang C and Li JC: Dexamethasone enhances trichosanthin-induced apoptosis in the HepG2 hepatoma cell line. Life Sci. 86:10–16. 2010. View Article : Google Scholar
12	Gottesman MM and Pastan I: Biochemistry of multidrug resistance mediated by the multidrug transporter. Annu Rev Biochem. 62:385–427. 1993. View Article : Google Scholar : PubMed/NCBI
13	Ambudkar SV, Dey S, Hrycyna CA, Ramachandra M, Pastan I and Gottesman MM: Biochemical, cellular and pharmacological aspects of the multidrug transporter. Annu Rev Pharmacol Toxicol. 39:361–398. 1999. View Article : Google Scholar
14	Ruefli AA and Johnstone RW: A role for P-glycoprotein in regulating cell growth and survival. Clin Appl Immunol Rev. 4:31–47. 2003. View Article : Google Scholar
15	Ruefli AA, Tainton KM, Darcy PK, Smyth MJ and Johnstone RW: P-glycoprotein inhibits caspase-8 activation but not formation of the death inducing signal complex (disc) following fas ligation. Cell Death Differ. 9:1266–1272. 2002. View Article : Google Scholar : PubMed/NCBI
16	Tainton KM, Smyth MJ, Jackson JT, Tanner JE, Cerruti L, Jane SM, Darcy PK and Johnstone RW: Mutational analysis of P-glycoprotein: Suppression of caspase activation in the absence of ATP-dependent drug efflux. Cell Death Differ. 11:1028–1037. 2004. View Article : Google Scholar : PubMed/NCBI
17	Pallis M, Turzanski J, Grundy M, Seedhouse C and Russell N: Resistance to spontaneous apoptosis in acute myeloid leukaemia blasts is associated with p-glycoprotein expression and function, but not with the presence of FLT3 internal tandem duplications. Br J Haematol. 120:1009–1016. 2003. View Article : Google Scholar : PubMed/NCBI
18	Narang VS, Fraga C, Kumar N, Shen J, Throm S, Stewart CF and Waters CM: Dexamethasone increases expression and activity of multidrug resistance transporters at the rat blood-brain barrier. Am J Physiol Cell Physiol. 295:C440–C450. 2008. View Article : Google Scholar : PubMed/NCBI
19	Chan GN, Saldivia V, Yang Y, Pang H, de Lannoy I and Bendayan R: In vivo induction of P-glycoprotein expression at the mouse blood-brain barrier: An intracerebral microdialysis study. J Neurochem. 127:342–352. 2013. View Article : Google Scholar : PubMed/NCBI
20	Rigalli JP, Perdomo VG, Luquita MG, Villanueva SS, Arias A, Theile D, Weiss J, Mottino AD, Ruiz ML and Catania VA: Regulation of biotransformation systems and ABC transporters by benznidazole in HepG2 cells: Involvement of pregnane X-receptor. PLoS Negl Trop Dis. 6:e19512012. View Article : Google Scholar : PubMed/NCBI
21	Jigorel E, Le Vee M, Boursier-Neyret C, Parmentier Y and Fardel O: Differential regulation of sinusoidal and canalicular hepatic drug transporter expression by xenobiotics activating drug-sensing receptors in primary human hepatocytes. Drug Metab Dispos. 34:1756–1763. 2006. View Article : Google Scholar : PubMed/NCBI
22	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar
23	Takeba Y, Sekine S, Kumai T, Matsumoto N, Nakaya S, Tsuzuki Y, Yanagida Y, Nakano H, Asakura T, Ohtsubo T and Kobayashi S: Irinotecan-induced apoptosis is inhibited by increased P-glycoprotein expression and decreased p53 in human hepatocellular carcinoma cells. Biol Pharm Bull. 30:1400–1406. 2007. View Article : Google Scholar : PubMed/NCBI
24	Sakaeda T, Nakamura T, Hirai M, Kimura T, Wada A, Yagami T, Kobayashi H, Nagata S, Okamura N and Yoshikawa T: MDR1 up-regulated by apoptotic stimuli suppresses apoptotic signaling. Pharm Res. 19:1323–1329. 2002. View Article : Google Scholar : PubMed/NCBI
25	Salje K, Lederer K, Oswald S, Dazert E, Warzok R and Siegmund W: Effects of rifampicin, dexamethasone, St. John's Wort and thyroxine on maternal and foetal expression of Abcb1 and organ distribution of talinolol in pregnant rats. Basic Clin Pharmacol Toxicol. 111:99–105. 2012.PubMed/NCBI
26	Petropoulos S, Gibb W and Matthews SG: Effect of glucocorticoids on regulation of placental multidrug resistance phosphoglycoprotein (P-gp) in the mouse. Placenta. 31:803–810. 2010. View Article : Google Scholar : PubMed/NCBI
27	Nishimura M, Koeda A, Morikawa H, Satoh T, Narimatsu S and Naito S: Comparison of inducibility of multidrug resistance (MDR)1, multidrug resistance-associated protein (MRP)1 and MRP2 mRNAs by prototypical microsomal enzyme inducers in primary cultures of human and cynomolgus monkey hepatocytes. Biol Pharm Bull. 31:2068–2072. 2008. View Article : Google Scholar : PubMed/NCBI
28	Mark PJ, Augustus S, Lewis JL, Hewitt DP and Waddell BJ: Changes in the placental glucocorticoid barrier during rat pregnancy: Impact on placental corticosterone levels and regulation by progesterone. Biol Reprod. 80:1209–1215. 2009. View Article : Google Scholar : PubMed/NCBI
29	Yumoto R, Murakami T, Sanemasa M, Nasu R, Nagai J and Takano M: Pharmacokinetic interaction of cytochrome P450 3A-related compounds with rhodamine 123, a P-glycoprotein substrate, in rats pretreated with dexamethasone. Drug Metab Dispos. 29:145–151. 2001.PubMed/NCBI
30	Demeule M, Jodoin J, Beaulieu E, Brossard M and Béliveau R: Dexamethasone modulation of multidrug transporters in normal tissues. FEBS Lett. 442:208–214. 1999. View Article : Google Scholar : PubMed/NCBI
31	Maier A, Zimmermann C, Beglinger C, Drewe J and Gutmann H: Effects of budesonide on P-glycoprotein expression in intestinal cell lines. Br J Pharmacol. 150:361–368. 2007. View Article : Google Scholar
32	Katayama K, Noguchi K and Sugimoto Y: FBXO15 regulates P-glycoprotein/ABCB1 expression through the ubiquitin-proteasome pathway in cancer cells. Cancer Sci. 104:694–702. 2013. View Article : Google Scholar : PubMed/NCBI
33	Micuda S, Fuksa L, Mundlova L, Osterreicher J, Mokry J, Cermanova J, Brcakova E, Staud F, Pokorna P and Martinkova J: Morphological and functional changes in p-glycoprotein during dexamethasone-induced hepatomegaly. Clin Exp Pharmacol Physiol. 34:296–303. 2007. View Article : Google Scholar : PubMed/NCBI
34	Mizutani T and Hattori A: New horizon of MDR1 (P-glycoprotein) study. Drug Metab Rev. 37:489–510. 2005. View Article : Google Scholar : PubMed/NCBI