Article Data

  • Views 1625
  • Dowloads 143

Original Research

Open Access

ALTERATION OF CYP3A1 mRNA LEVEL IN PRIMARY RAT HEPATOCYTES IN RESPONSE TO AMPK ACTIVATOR AICAR  

  • Bang-Sub Lee1
  • Jooyoung Kim2
  • Wi-Young So1

1Sports and Health Care Major, College of Humanities and Arts, Korea National University of Transportation, Chungju-si, Republic of Korea

2Sports, Health, and Rehabilitation Major, College of Physical Education, Kookmin University, Seoul, Republic of Korea

DOI: 10.22374/jomh.v15i1.113 Vol.15,Issue 1,March 2019 pp.23-29

Published: 22 March 2019

*Corresponding Author(s): Wi-Young So E-mail: wowso@ut.ac.kr

Abstract

Background and Objective

AMP-activated protein kinase (AMPK) functions as a sensor of the intracellular energy status that can be stimulated by a synthetic activator, 5-aminoimidazole–4–carboxamide–1–beta–D–ribofuranoside (AICAR), which is used to replicate the effect of physical exercise in hepatocyte embryoid bodies. This study inves-tigated the effect of AICAR on the CYP3A1 mRNA expression in primary hepatocyte embryoid bodies derived from a rat liver.

Material and Methods

The primary hepatocytes were isolated from a male Sprague Dawley (SD) rat (215 g) and subjected to the following treatments: control without AICAR (CTL, n=3), 1 μM AICAR (n=3), 10 μM AICAR (n=3), and 100 μM AICAR (n=3). RNA was isolated and used as the template for synthesizing cDNA by reverse tran-scriptase to perform quantitative PCR (qPCR). The independent samples t-test was conducted to examine differences between groups. Statistical significance was set at p<0.05.

Results

The qPCR analysis demonstrated that CYP3A1 mRNA expression in primary hepatocyte embryoid bodies significantly increased in the presence of 10 μM (t=1.730, p<0.05) and 100 μM AICAR (t=3.207, p<0.05) as compared to that in the control group hepatocytes. However, the observed increase of CYP3A1 mRNA in hepatocyte embryoid bodies was not statistically significant in the presence of 1 μM AICAR as the low-est test concentration.

Conclusion

In this study, we demonstrated that AICAR, an AMPK activator, can increase the expression of CYP3A1 mRNA in primary hepatocytes. Future studies should assess the effect of AICAR treatment on CYP3A4 in human hepatocytes.

Keywords

5-aminoimidazole–4–carboxamide–1–beta–D–ribofuranoside, AMP-activated protein kinase, CYP3A1

Cite and Share

Bang-Sub Lee,Jooyoung Kim,Wi-Young So. ALTERATION OF CYP3A1 mRNA LEVEL IN PRIMARY RAT HEPATOCYTES IN RESPONSE TO AMPK ACTIVATOR AICAR . Journal of Men's Health. 2019. 15(1);23-29.

References

1. Merrill GF, Kurth EJ, Hardie DG, et al. AICA riboside increases AMP-activated protein kinase, fatty acid oxidation, and glucose uptake in rat muscle. Am J Physiol 1997;273(6 Pt 1):1107–12.

2. Chambers MA, Moylan JS, Smith JD, et al. Stretch-stimulated glucose uptake in skeletal muscle is medi-ated by reactive oxygen species and p38 MAP-kinase. J Physiol 2009;587(13):3363–73.

3. Witczak CA, Sharoff CG, Goodyear LJ. AMP-activated protein kinase in skeletal muscle: from structure and localization to its role as a master regulator of cellular metabolism. Cell Molec Life Scie 2008;65(23):3737–55.

4. Gibala MJ, McGee SL, Garnham AP, et al. Brief intense interval exercise activates AMPK and p38 MAPK signaling and increases the expression of PGC-1alpha in human skeletal muscle. J Appl Physiol 1985;106(3):929–34.

5. Williamson DL, Bolster DR, Kimball SR, et al. Time course changes in signaling pathways and protein syn-thesis in C2C12 myotubes following AMPK activation by AICAR. American journal of physiology. Endocrin Metab 2006;291(1):80–89.

6. Guengerich FP, Shimada T, Yun CH, et al. Interactions of ingested food, beverage, and tobacco components involving human cytochrome P4501A2, 2A6, 2E1, and 3A4 enzymes. Environ Health Perspect 1994;9:49–53.

7. Stupans I, Murray M, Kirlich A, et al. Inactivation of cytochrome P450 by the food-derived complex phenol oleuropein. Food Cheml Toxicol 2001;39(11):1119–24.

8. Martignoni M, Groothuis G, de Kanter R. Com-parison of mouse and rat cytochrome P450-mediated metabolism in liver and intestine. Drug Metabol Disposit 2006;34(6):1047–54.

9. Furge LL, Guengerich FP. Cytochrome P450 enzymes in drug metabolism and chemical toxicology: An introduc-tion. Biochem Molec Biol Educat 2006;34(2):66–74.

10. Smith DA, Abel SM, Hyland R, et al. Human cy-tochrome P450s: selectivity and measurement in vivo. Xenobiotica 1998;28(12):1095–28.

11. Wang P, Mason PS, Guengerich FP. Purification of human liver cytochrome P-450 and comparison to the enzyme isolated from rat liver. Arch Biochem Biophys 1980;199(1):206–19.

12. Frenkl R, Szeberenyi S. Enzyme inducing effect of muscular exertion in the rat. Acta Medica Academ Scient Hungaricae 1976;33(1):95–100.

13. Ware WR. Nutrition and the prevention and treatment of cancer: association of cytochrome P450 CYP1B1 with the role of fruit and fruit extracts. Integrat Cancer Ther 2009;8(1):22–28.

14. Guengerich FP. Cytochrome P-450 3A4: regulation and role in drug metabolism. Ann Rev Pharmacol Toxicol 1999;39:1–17.

15. Watkins PB, Wrighton SA, Schuetz EG, et al. Identifi-cation of glucocorticoid-inducible cytochromes P-450 in the intestinal mucosa of rats and man. J Clin Invest 1987;80(4):1029–36.

16. Lown KS, Bailey DG, Fontana RJ, et al. Grapefruit juice increases felodipine oral availability in humans by decreasing intestinal CYP3A protein expression. J Clin Investig 1997;99(10):2545–53.

17. Kolars JC, Lown KS, Schmiedlin-Ren P, et al. CYP3A gene expression in human gut epithelium. Pharmaco-genetics 1994;4(5):247–59.

18. Huang Z, Fasco MJ, Figge HL, et al. Expression of cytochromes P450 in human breast tissue and tumors. Drug Metab Disposit 1996;24(8):899–905.

19. Shimada T, Yamazaki H, Mimura M, et al. Interindividual variations in human liver cytochrome P-450 enzymes involved in the oxidation of drugs, carcinogens and toxic chemicals: studies with liver microsomes of 30 Japanese and 30 Caucasians. J Pharmacol Experiment Ther 1994;270(1):414–23.

20. Takizawa D, Kakizaki S, Horiguchi N, et al. Constitutive active/androstane receptor promotes hepatocarcinogen-esis in a mouse model of non-alcoholic steatohepatitis. Carcinogenesis 2011;32(4):576–83.

21. Cuciureanu M, Vlase L, Muntean D, et al. Grapefruit juice--drug interactions: importance for pharmacotherapy. Rev Med Chir Soc Med Nat Iasi 2010;114(3):885–91.

22. Schmucker DL, Ohta M, Kanai S, et al. Hepatic injury induced by bile salts: correlation between biochemical and morphological events. Hepatology 1990;12(5):1216–21.

23. Jorquera F, Almar MM, Pozuelo M, et al. Effects of aging on antipyrine clearance: predictive factors of metaboliz-ing capacity. J Clin Pharmacol 1995;35(9):895–901.

24. Paterson DH, Jones GR, Rice CL. Ageing and physical activity: evidence to develop exercise recommendations for older adults. Can J Public Health 2007;98(Suppl 2):S69–108.

25. Liptrott NJ, Penny M, Bray PG, et al. The impact of cytokines on the expression of drug transporters, cytochrome P450 enzymes, and chemokine receptors in human PBMC. Br J Pharmacol 2009;156(3):497–8.

26. Wolbold R, Klein K, Burk O, et al. Sex is a major determinant of CYP3A4 expression in human liver. Hepatology 2003;38(4):978–88.

27. Tanaka E. In vivo age-related changes in hepatic drug-oxidizing capacity in humans. J Clin Pharm Ther 1998;23(4):247–55.

28. Badyal DK, Dadhich AP. Cytochrome P450 and drug interactions. Indian J Pharmacol 2001;33(4):248–59.

29. Schwartz JB. The influence of sex on pharmacokinet-ics. Clin Pharmacokinet 2003;42(2):107–21.

30. Harris RZ, Benet LZ, Schwartz JB. Gender effects in pharmacokinetics and pharmacodynamics. Drugs 1995;50(2):222–39.

31. Meibohm B, Beierle I, Derendorf H. How important are gender differences in pharmacokinetics? Clin Pharmacokinet 2002;41(5):329–42.

32. Lew KH, Ludwig EA, Milad MA, et al. Gender-based effects on methylprednisolone pharmacokinet-ics and pharmacodynamics. Clin Pharmacol Ther 1993;54(4):402–14.

33. Meffin PJ, Brooks PM, Sallustio BC. Alterations in prednisolone disposition as a result of time of ad-ministration, gender and dose. Br J Clin Pharmacol 1984;17(4):395–4.

34. Fukuno S, Nagai K, Horii A et al. Pharmacokinetics and metabolic elimination of tolbutamide in female rats: Comparison with male rats. Biopharm Drug Dispos 2018;39(7):321–27.

35. Pritchard JF, Bryson JC, Kernodle AE, et al. Age and gender effects on ondansetron pharmacokinetics: evaluation of healthy aged volunteers. Clin Pharmacol Ther 1992;51(1):51–55.

36. Kobayashi K, Abe C, Endo M, et al. Gender Difference of hepatic and intestinal CYP3A4 in CYP3A humanized mice generated by a human chromosome-engineering technique. Drug Metab Lett 2017;11(1):60–67.

37. Staskin DR. Overactive bladder in the elderly: a guide to pharmacological management. Drugs Aging 2005;22(12):1013–28.

38. De Stefano F, Zambon S, Giacometti L, et al. Obesity, muscular strength, muscle composition and physical performance in an elderly population. J Nutrition Health Aging 2015;19(7):785–91.

39. Wauthier V, Verbeeck RK, Calderon PB. The effect of ageing on cytochrome p450 enzymes: consequences for drug biotransformation in the elderly. Curr Med Chem 2007;14:745–57.

40. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T) method. Methods 2001;25(4):402–8.

41. Lee CM, Pohl J, Morgan ET. Dual mechanisms of CYP3A protein regulation by proinflammatory cy-tokine stimulation in primary hepatocyte cultures. Drug Metabol Disposit 2009;37(4):865–72.

42. Hu N, Hu M, Duan R, et al. Increased levels of fatty acids contributed to induction of hepatic CYP3A4 activity induced by diabetes - in vitro evidence from HepG2 cell and Fa2N-4 cell lines. J Pharmacol Sci 2014;124(4):433–44.

43. 43. Vignati L, Turlizzi E, Monaci S, et al. An in vitro approach to detect metabolite toxicity due to CYP3A4 dependent bioactivation of xenobiotics. Toxicology 2005;216(2-3):154–67.

44. van Herwaarden AE, Wagenaar E, van der Kruijssen CM, et al. Knockout of cytochrome P450 3A yields new mouse models for understanding xenobiotic metabolism. J Clin Investigat 2007;117(11):3583–92.

45. Lee BS, So WY, Kim SH, et al. Expression profiles of cytochrome P450s following swimming exercise in aging rats. J Men’s Health 2017;13(2):e25–e33.

46. Lee BS, So WY, Chung WY. Suppression of expression levels of constitutive androstane receptor by moderate exercise in BALB/c nude mice with breast cancer. Iran J Pub Health 2017;46(8):1154–55.

47. Jaffe CA, Ocampo-Lim B, Guo W, et al. Regulatory mechanisms of growth hormone secretion are sexu-ally dimorphic. J Clin Investig 1998;102(1):153–64.

48. Park SH, Liu X, Hennighausen L, et al. Distinctive roles of STAT5a and STAT5b in sexual dimorphism of hepatic P450 gene expression: impact of STAT5a gene disruption. J Biol Chem 1999;274(11):7421–30.

49. Waxman DJ, LeBlanc GA, Morrissey JJ, et al. Adult male-specific and neonatally programmed rat hepatic 

P- 450 forms RLM2 and 2a are not dependent on pul-satile plasma growth hormone for expression. J Biol Chem 1988;263(23):11396–406.

50. Robertson GR, Farrell GC, Liddle C. Sexually dimor-phic expression of rat CYP3A9 and CYP3A18 genes is regulated by growth hormone. Biochem Biophys Res Commun 1998;242(1):57–60.

51. Kawai M, Bandiera SM, Chang TK, et al. Growth hormone regulation and developmental expression of rat hepatic CYP3A18, CYP3A9 and CYP3A2. Biochem Pharmacol 2000;59(10):1277–87.

52. Sakuma T, Endo Y, Mashino M, et al. Regulation of the expression of two female-predominant CYP3A mRNAs (CYP3A41 and CYP3A44) in mouse liver by sex and growth hormones. Arch Biochem Biophys 2002;40492:234–42.

53. Cheung C, Yu AM, Chen CS, et al. Growth hormone determines sexual dimorphism of hepatic cytochrome P450 3A4 expression in transgenic mice. J Pharmacol Exp Ther 2006;316(3):1328–34.

54. van den Berg G, Veldhuis JD, Frolich M, et al. An amplitude-specific divergence in the pulsatile mode of growth hormone (GH) secretion underlies the gender difference in mean GH concentrations in men and premenopausal women. J Clin Endocrinol Metab 1996;81(7):2460–67.

55. Hashimoto M, Kobayashi K, Watanabe M, et al. Knockout of mouse Cyp3a gene enhances synthesis of cholesterol and bile acid in the liver. J Lipid Res 2013;54(8):2060–68.

Abstracted / indexed in

Science Citation Index Expanded (SciSearch) Created as SCI in 1964, Science Citation Index Expanded now indexes over 9,200 of the world’s most impactful journals across 178 scientific disciplines. More than 53 million records and 1.18 billion cited references date back from 1900 to present.

Journal Citation Reports/Science Edition Journal Citation Reports/Science Edition aims to evaluate a journal’s value from multiple perspectives including the journal impact factor, descriptive data about a journal’s open access content as well as contributing authors, and provide readers a transparent and publisher-neutral data & statistics information about the journal.

Directory of Open Access Journals (DOAJ) DOAJ is a unique and extensive index of diverse open access journals from around the world, driven by a growing community, committed to ensuring quality content is freely available online for everyone.

SCImago The SCImago Journal & Country Rank is a publicly available portal that includes the journals and country scientific indicators developed from the information contained in the Scopus® database (Elsevier B.V.)

Publication Forum - JUFO (Federation of Finnish Learned Societies) Publication Forum is a classification of publication channels created by the Finnish scientific community to support the quality assessment of academic research.

Scopus: CiteScore 0.7 (2022) Scopus is Elsevier's abstract and citation database launched in 2004. Scopus covers nearly 36,377 titles (22,794 active titles and 13,583 Inactive titles) from approximately 11,678 publishers, of which 34,346 are peer-reviewed journals in top-level subject fields: life sciences, social sciences, physical sciences and health sciences.

Norwegian Register for Scientific Journals, Series and Publishers Search for publication channels (journals, series and publishers) in the Norwegian Register for Scientific Journals, Series and Publishers to see if they are considered as scientific. (https://kanalregister.hkdir.no/publiseringskanaler/Forside).

Submission Turnaround Time

Conferences

Top