DOI QR코드

DOI QR Code

Effect of Hormones and Short Chain Fatty Acids on CYP7A1 Gene Expression in HepG2 Cell

호르몬과 단쇄지방산이 HepG2 Cell 내에서 CYP7A1 발현에 미치는 효과

  • Yang, Jeong-Lye (Research Institute Of Human Ecology, Changwon University) ;
  • Lee, Hyun-Jung (Dept. of Food and Nutritional Sciences, Ewha Womans University) ;
  • Kim, Yang-Ha (Dept. of Food and Nutritional Sciences, Ewha Womans University)
  • 양정례 (창원대학교 생활과학연구소) ;
  • 이현정 (이화여자대학교 식품영양학과) ;
  • 김양하 (이화여자대학교 식품영양학과)
  • Published : 2005.06.01

Abstract

Cholesterol $7\alpha-hydroxylase$ (CYP7A1) is the rate-limiting enzyme in the conversion of cholesterol to bile acids and plays a central role in regulating cholesterol homeostasis. We previously showed that a fermentable $\beta-glucan$ ingestion decreased plasma cholesterol levels due to fecal bile acid excretion elevation involved inincrease of cholesterol $7\alpha-hydroxylase$ mRNA expression and activity. It is proposed that short chain fatty acids (SCFA) produced by cecal and colonic fermentation of soluble fiber are associated with cholesterol-lowering effect of fiber. In the present study, we investigated whether CYP7A1 expression is up-regulated by short chain fatty acids or by hormones in cultured human hepatoma (HepG2) cells. Confluent HepG2 cell were incubated with acetate, propionate, or butyrate at 1 mM concentration for 24 hrs. Acetate as well as propionate increased to 1.8-fold expression of CYP7A1 mRNA than the control. Butyrate also increased 1.5-fold expression of CYP7A1 mRNA. Our data show for the first time that SCFA increase expression of CYP7A1 mRNA. Adding insulin, dexamethasone and triiodothyronine $(1\;{\mu}M)$ to HepG2 cell increased the expression of CYP7A1 mRNA to $150\%,\;173\%,\;141\%$, respectively. These results suggest that SCFA produced by cecal fermentation stimulate enteric nervous system, in which secreted some neuropeptides may be responsible for change in cholesterol and bile acid metabolism. These findings suggest that SCFA are involved in lowering plasma cholesterol levels due to the up-regulation of CYP7A1 and bile acid synthesis.

References

  1. Vlahcevic ZR, Heuman DM, Hylemon PB. 1991. Regulation of bile acid synthesis. Hepatology 13: 590-600 https://doi.org/10.1002/hep.1840130331
  2. Pandak WM, Li YC, Chiang JYL, Studer EJ, Gurley EC, Heuman DM, Hylemon PB. 1991. Regulation of cholesterol 7$alpha$-hydroxyalse mRNA and transcriptional activity by taurocholate and cholesterol in the chronic biliary diverted rat. J Biol Chem 266: 3416-3421
  3. Li YC, Wang DP, Chiang JYL. 1990. Regulation of cholesterol 7$alpha$-hydroxylase in the liver. J Biol Chem 265: 12012-12019
  4. Jelinek DF, Andersson S, Slaughter CA, Russell DW. 1990. Cloning and regulation of cholesterol 7$alpha$-hydroxylase, the rate-limiting in bile acid biosynthesis. J Biol Chem 265: 8190-8197
  5. Ness GC, Pendleton LC, Li YC, Chiang JYL. 1990. Effect of thyroid hormone on hepatic cholesterol 7$alpha$-hydroxylase, LDL receptor, HMG-CoA reductase, famesyl pyrophosphate synthetase and apolipoprotein A-I mRNA levels in hypophysectomized rats. Biochem Biophys Res Commun 172: 1150-1156 https://doi.org/10.1016/0006-291X(90)91568-D
  6. Hylernon PB, Gurley EC, Stravitz RT, Litz JS, Pandak WM, Chiang JYL, Vlahcevic ZR. 1992. Hormonal regulation of cholesterol 7$alpha$-hydroxylase mRNA levels and transcriptional activity in primary rat hepatocyte cultures. J Biol Chem 267: 16866-16871
  7. Twisk JM, Hoekman FM, Lehmann EM, Meijer P, Mager WH, Princen HMG. 1995. Insulin suppresses bile acid synthesis in cultured rat hepatocytes by down-regulation of cholesterol 7$alpha$-hydroxylase and sterol 27-hydroxylase gene transcription. Hepatology 21: 501-510
  8. Lavery DJ, Schibler U. 1993. Circadian transcription of the cholesterol 7$alpha$ -hydroxylase gene may involve the liverenriched bZIP protein DBP. Gene & Dev 7: 1871-1884 https://doi.org/10.1101/gad.7.10.1871
  9. Jelinek D, Russel DW. 1990. Structure of the rat gene encoding cholesterol 7 $alpha$-hydroxylase. Biochemistry 29: 7781-7785 https://doi.org/10.1021/bi00486a001
  10. Nishimoto M, Gotoh O, Okuda K, Noshiro M. 1991. Structural anaylsis of the gene encoding rat cholesterol 7$\alpha$-hydroxylase, the key enzyme for bile acid synthesis. J Biol Chem 266: 6467-6471
  11. Chiang JYL, Yang TP, Wang DP. 1992. Cloning and 5'-flanking sequence of a rat cholesterol 7$alpha$-hydroxylase gene. Biochim Biophys Acta 1132: 337-339 https://doi.org/10.1016/0167-4781(92)90175-Y
  12. Molowa DT, Chen WS, Cimis GM, Tan CP. 1992. Transcriptional regulation of the human cholesterol 7$\alpha$-hydroxylase gene. Biochemistry 31: 2539-2544 https://doi.org/10.1021/bi00124a014
  13. Cohen JC, Cali JJ, Jelinek DF, Mehrabian M, Sparkes RS, Lusis AJ, Russell DW, Hobbs HH. 1992. Cloning of the human cholesterol 7$alpha$-Iiydroxylase gene (CYP7) and localization to chromosome 8q11-q12. Genomics 14: 153-161 https://doi.org/10.1016/S0888-7543(05)80298-8
  14. Wang DP, Chiang JYL. 1994. Structure and nucleotide sequences of the human cholesterol 7$alpha$-hydroxylase gene (CYP7). Genomics 20: 320-323 https://doi.org/10.1006/geno.1994.1177
  15. Crestani M, Galli G, Chaing JYL. 1993. Genomic cloning, sequencing and analysis of hamster cholesterol 7$alpha$-hydroxylase gene (CYP7). Arch Biochem Biophys 306: 451-460 https://doi.org/10.1006/abbi.1993.1537
  16. Tzung K, Ishimura-Oka K, Kihara S, Oka K, Chan L. 1994. Structure of the mouse cholesterol 7$alpha$-Iiydroxylase gene. Genomics 21: 244-247 https://doi.org/10.1006/geno.1994.1250
  17. Davis RA, Highsmith WE, Malone-McNeal M, ArchambaultSchexnayder J, Kuan JCW. 1983. Bile acid synthesis by cultured hepatocytes: inhibition by mevinolin but not by bile acids. J Biol Chem 258: 4079-4082
  18. Stravitz RT, Hylemon PB, Heuman DM, Hagey LR, Schteingart CD, Ton-Nu H-T, Hoffman AF, Vlahcevic ZR. 1993. Transcriptional regulation of cholesterol 7 $alpha$-hydroxylase mRNA by conjugated bile acids in primary cultures of rat hepatocytes. J Biol Chem 268: 13987-13993
  19. Kwekkeboom J. Princen HMG, Van Voorthuizen EM, Kempen HJM. 1990. Bile acids exert negative feedback control on bile acid synthesis in cultured pig hepatocytes by suppression of cholesterol 7$\alpha$-hydroxylase, Hepatology 12: 1209-1215 https://doi.org/10.1002/hep.1840120522
  20. Everson GT, Polokoff MA. 1986. A human hepatomablastoma line exhibiting defects in bile acid synthesis and conjugation. J Biol Chem 261: 2197-2201
  21. Yang JL, Kim YH, Lee HS, Lee MS, Moon YK. 2003. Barley beta-glucan lowers serum cholesterol 7alpha-hydroxylase activity and mRNA abundance in cholesterol-fed rats. J Nutr Sci Vitaminol 49: 381-387 https://doi.org/10.3177/jnsv.49.381
  22. Anderson JW. 1985. Physiological and metabolic effects of dietary fiber. Fed Proc 44: 2902-2906
  23. Moundras C, Demigne C, Mazur A, Remesy C. 1995. The cholesterol lowering effect of steroid sequestrants is modulated by large intestine fermentation. J Nutr Biochem 6: 158-162 https://doi.org/10.1016/0955-2863(94)00016-F
  24. Nishimura N, Nishikawa H, Kiriyama S. 1993. Ileorectostomy or cecectomy but not colectomy abolishes the plasma cholesterol-lowering effect of dietary beet fiber in rats. J Nutr 123: 1260-1269
  25. Hara H, Haga S, Takanori K, Kiriyama S. 1998. Fermentation products of sugar-beet fiber by cecal bacteria lower plasma cholesterol concentration in rats. J Nutr 128: 688-693
  26. Hara H, Haga S, Aoyama Y, Kiriyama S. 1999. Short-chain fatty acids suppress cholesterol synthesis in rat liver. J Nutr 129: 942-948
  27. Lin Y, Vonk RJ, Slooff MJ, Kuipers F, Smit MJ. 1995. Differences in propionate-induced inhibition of cholesterol and triacylglycerol synthesis between human and rat hepatocytes in primary culture. Br J Nutr 74: 197-207 https://doi.org/10.1079/BJN19950123
  28. Nishina PM, Freedl RA. 1990. Effects of propionate on lipid biosynthesis in isolated rat hepatocytes. J Nutr 120: 668-673 https://doi.org/10.1093/jn/120.7.668
  29. Goodlad RA, Ratcliffe B, Fordham JP, Ghatei MA, Domin J, Bloom SR, Wright NA. 1989. Plasma enteroglucagon, gastrin and peptide YY in conventional and germ-free rats refed with a fiber-free or fiber-supplemented diet. Q J Exp Physiol 74: 437-442
  30. Kennedy HJ, Sarson DL, Bloom SR, Truelove SC. 1982. Gut hormone responses in subjects with a permanent ileosstorny. Digestion 24: 133-136 https://doi.org/10.1159/000198788
  31. Cho WK, Mennone A, Rydberg SA, Boyer JL. 1997. Bombesin stimulates bicarbonate secretion from rat cholangiocytes: implications for neural regulation of bile secretion. Gastroenterology 113: 311-321 https://doi.org/10.1016/S0016-5085(97)70109-4
  32. Farouk M, Geoghegan JG, Pruthi RS, Thomson HJ, Pappas TN, Meyers WC. 1992. Intracerebroventricular neuropeptide Y stimulates bile secretion via a vagal mechanism. Gut 33: 1562-1565 https://doi.org/10.1136/gut.33.11.1562
  33. Gebhard RL, Morley JE, Prigge WF, Goodman MW, Prasad C. 1981. TRH and histidyl-proline diketopiperazine inhibit cholesterol synthesis in dog intestine. Peptides 2: 137-140
  34. Levy-Wilson B, Fortier C, Blackhart BD, McCarthy BJ. 1988. DNase I -and micrococcal nuclease-hypersensitive sites in the human apolipoprotein B gene are tissue specific. Mol Cell Biol 8: 71-80 https://doi.org/10.1128/MCB.8.1.71
  35. Pohlodek K, Galbavy S, Bartosova M, Mucha V, Holoman K, Pastorek J, Pastorekova S. 2004. Semi-quantitative RTPCR assessment of molecular markers in breast large-core needle biopsies. Neoplasma 51: 415-421
  36. Twisk J, Lehmann EM, Princen HMG. 1993. Differential feedback regulation of cholesterol 7 alpha-hydroxylase mRNA and transcriptional activity by rat bile acids in primary monolayer cultures of rat hepatocytes. Biochem J 290: 685-691 https://doi.org/10.1042/bj2900685
  37. Princen HMG, Meijer P, Hofstee B. 1989. Dexamethasone regulates bile acid synthesis in monolayer cultures of rat hepatocytes by induction of cholesterol 7 alpha-hydroxylase. Biochem J 262: 341-348 https://doi.org/10.1042/bj2620341
  38. Lavery DJ, Schibler U. 1993. Circadian transcription of the cholesterol 7 alpha-hydroxylase gene may involve the liver-enriched bZIP protein DBP. Genes Dev 7: 1871-1884 https://doi.org/10.1101/gad.7.10.1871
  39. Rudel L, Deckelman C, Wilson M, Scobey M, Anderson R. 1994. Dietary cholesterol and down-regulation of cholesterol 7$alpha$-hydroxyalse and cholesterol absorption in African green monkeys. J Clin Invest 93: 2463-2472 https://doi.org/10.1172/JCI117255
  40. Poorman JA, Buck RA, Smith SA, Overtunf ML, LooseMitchell DS. 1993. Bile acid excretion and cholesterol 7$alpha$-hydroxylase expression in hypercholesterolemia-resistant rabbits. J Lipid Res 34: 1675-1685
  41. Bertolotti M, Abate N, Loria P, Dilengite M, Carubbi F, Pinetti A, Digrisolo A, Carulli N. 1991. Regulation of bile acid synthesis in humans: effect of treatment with bile acids, cholestyramine, or simvastatin on cholesterol 7$alpha$- hydroxylase rats in vivo. Hepatology 14: 830-837 https://doi.org/10.1002/hep.1840140515
  42. Karam WG, Chiang JYL. 1994. Expression and purification of human cholesterol 7$alpha$-hydroxylase in Escherichia coli. J Lipid Res 35: 1222-1231
  43. Cooper AD, Craig WY, Taniguchi T, Everson GT. 1994. Characteristics and regulation of bile salts synthesis and secretion by human hepatoma HepG2 cells. Hepatology 20: 1522-1531 https://doi.org/10.1002/hep.1840200623
  44. Anderson JW, Deakins DA, Floore TL, Smith BM, Whitis SE. 1990. Dietary fiber and coronary heart disease. Crit Rev Food Sci Nutr 29: 95-147 https://doi.org/10.1080/10408399009527518
  45. Jackson KA, Sute DA, Topping DL. 1994. Oat bran, barley and malted barley lower plasma cholesterol relative to wheat bran but differ in their effects on liver cholesterol in rats fed diets with and without cholesterol. J Nutr 124: 1678-1684
  46. IIlman RJ, Trimble RP, Snoswell AM, Topping DL. 1982. Daily variations in the concentrations of volatile fatty acids in the blood vessels of rats fed diets high in pectin and bran. Nutr Rep Int 26: 439-446
  47. Nishina PM, Freedland RA. 1990. Effects of propionate on lipid biosynthesis in isolated rat hepatocytes. J Nutr 120: 668-673 https://doi.org/10.1093/jn/120.7.668
  48. Anderson JW, Bridges SR. 1984. Short-chain fatty acid fermentation products of plant fiber affect glucose metabolism of isolated rat hepatocytes. Proc Soc Exp Biol Med 177: 372-376
  49. Chen WJ, Anderson JW, jennings D. 1984. Propionate may mediate the hypocholesterolemic effects of certain soluble plant fibers in cholesterol-fed rats. Proc Soc Exp Biol Med 175: 215-218
  50. Wright RS, Anderson JW, Bridges SR. 1990. Propionate inhibits hepatocyte lipid synthesis. Proc Soc Exp Biol Med 195: 26-29
  51. Demigne C, Mor C, Levrat MA, Besson C, Moundras C, Remesy C. 1995. Effect of propionate on fatty acid and cholesterol synthesis and on acetate metabolism in isolated rat hepatocytes. Br J Nutr 74: 209-219 https://doi.org/10.1079/BJN19950124
  52. Kishimoto Y, Wakabayashi S, Takeda H. 1995. Effects of intravenous injection and intraperitoneal continual administration of sodium propionate on serum cholesterol levels in rats. J Nutr Sci Vitaminol 41: 73-81 https://doi.org/10.3177/jnsv.41.73
  53. Levrat MA, Favier ML, Moundras C, Remesy C, Demigne C, Morand C. 1994. Role of dietary propionic acid and bile acid excretion in the hypocholesterolemic effects of oligosaccharides in rats. J Nutr 124: 531-538
  54. Beaulieu KE, McBurney MI. 1992. Changes in pig serum lipids, nutrient digestibility and sterol excretion during cecal infusion of propionate. J Nutr 122: 241-245 https://doi.org/10.1093/jn/122.2.241
  55. Arjmi BH, Craig J, Nathani S, Reeves RD. 1992. Soluble dietary fiber and cholesterol influence in vivo hepatic and intestinal cholesterol biosynthesis in rats. J Nutr 122: 1559-1565 https://doi.org/10.1093/jn/122.7.1559
  56. Stark AH, Madar Z. 1993. In vitro production of short-chain fatty acids by bacterial fermentation of dietary fiber compared with effects of those fibers on hepatic sterol synthesis in rats. J Nutr 123: 2166-2173
  57. Illman RJ, Topping DL. 1985. Effects of dietary oat bran on fecal steroid excretion, plasma volatile fatty acids and lipid synthesis in rats. Nutr Res 5: 839-846 https://doi.org/10.1016/S0271-5317(85)80171-8
  58. Illman RJ, Topping DL, Mcintosh GH, Trimble RP, Storer GB, Taylor MN, Cheng BQ. 1988. Hypocholesterolemic effects of dietary propionate: studies in whole animals and perfused rat liver. Ann Nutr Metab 32: 95-107

Cited by

  1. Effect of Artemisia capillaries on Gene Expression of Lipid Metabolism in Rat vol.14, pp.3, 2011, https://doi.org/10.3831/KPI.2011.14.3.063
  2. Transcriptomic and metabolomic responses induced in the livers of growing pigs by a short-term intravenous infusion of sodium butyrate vol.12, pp.11, 2018, https://doi.org/10.1017/S1751731118000174