Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Anti-obesity and Anti-inflammation Effects of Cheonggukjang in C57Bl/6 mice with High Fat Diet Induced Obesity

고지방식이로 유도된 비만 마우스에서 청국장의 항비만 및 항염증 효과

  • Kim, Jiyoung (Departments of Food Science and Nutrition Pusan National University)
  • 김지영 (부산대학교 식품영양학과)
  • Received : 2017.11.10
  • Accepted : 2017.11.20
  • Published : 2017.11.30
Download PDF
⟨ Previous Next ⟩

Abstract

The purpose of this study was to investigate the anti-obesity and anti-inflammation effect of the cheonggukjang (a soybean paste fermented for only a few days) in diet induced obesity mice. Weight gain was significantly decreased in the mice fed cheonggukjang compared High Fat Diets (HFD). The HFD plus cheonggukjang (CGJ) were also effective in improving the lipid metabolism. The levels of plasma triglyceride, cholesterol, ALT, AST, leptin, glucose, and insulin were significantly lower in CGJ than HFD group (p<0.05). The adiponectin level of CGJ group was significantly increased compared to the HFD group (p<0.05). In the CGJ group, the mRNA expression of adipogenic genes in the liver and adipose tissues, which are transcription factors crucial for adipogenesis, were significantly suppressed (p<0.05). The number of $CD11b^+F4/80^+$ T cells, $Gr-1^{int}CD11b^{high}$ cells, and $Gr-1^{int}CD11b^{high}$ cells were significantly higher in HFD group than CGJ group (p<0.05). The size of adipocyte was significantly reduced in CGJ group compared to HFD group. In addition, the contents of liver lipid droplets were significantly downregulated in the CGJ mice than HFD mice (p<0.05). Collectively, these data suggest the novel function of cheonggukjang in modulating adipogenesis through an immune function-alteration involving downregulation of adipogenic transcription factors and macrophage activation.

본 연구에서는 청국장의 항비만 및 항염증 효과를 알아보고자 C57BL/6 마우스에 고지방식이를 13주감 섭취시켜 비만을 유도한 후에 고지방 식이에 청국장을 10% 첨가하여 항비만과 항염증 활성을 확인하였다. 고지방식이를 통해 비만이 유도된 mice (HFD)에 고지방식이와 함께 청국장을 첨가하여 섭취한 그룹(CGJ)의 체중 증가율은 고지방식이군과 비교하였을 때 현저하게 감소하였을 뿐만 아니라 정상식이군보다도 유의적으로 감소하였으며 혈중 TG, TC, glucose, insulin, AST와 ALT 분비는 감소하였고 adiponectin의 수치는 유의적으로 증가하였다(p<0.05). 또한 H&E와 Oil red O staining을 통해 13주 동안 고지방식이를 통해 유도된 지방 간의 심각정도가 청국장의 섭취로 인하여 감소한 것을 확인하였다(p<0.05). 간세포는 대식세포, T세포와 같은 면역시스템의 세포들을 포함하고 있는데 비만으로 유도된 간 조직의 염증반응에 청국장이 미치는 영향을 조사하기 위해 FACS 분석을 수행하였다. CGJ군은 HFD군에 비하여 T cell의 침윤과 대식세포의 축적, 호중구의 유입이 유의적으로 감소하였다(p<0.05). 간 조직으로 침윤된 면역세포의 증가가 간 조직 내 염증반응을 유도하는지 확인하기 위해 염증성 cytokine의 수준을 조사한 결과 간과 지방 조직에서 염증성 cytokine인 MCP-1과 $TNF-{\alpha}$의 mRNA 발현이 감소하였으며 지방합성에 관여하는 $PPAR-{\gamma}$, SREBP-1와 $CEBP/{\alpha}$의 mRNA 발현 역시 감소하였다. 이러한 결과들을 통해 청국장이 면역 시스템과 지방생성 전사 인자들의 발현에 관여함으로써 adipogenesis를 조절함과 동시에 염증 억제 효과를 가진다는 사실을 증명하였다.

Keywords

References

  1. Akram, D. S. Report of WHO Consultation on Obesity, 2000, WHO: Obesity: Preventing and Managing the Global Epidemic, pp.6-8, World Health Organization, Geneva, Switzerland
  2. Aoki, F. and Honda, H. 2007. Suppression by Licorice Flavonoids of Abdominal Fat Accumulation and Body Weight Gain in High-Fat Diet-Induced Obese C57BL/6 Mice. Biosci. Biotechnol. Biochem. 71, 206-214. https://doi.org/10.1271/bbb.60463
  3. Bechmann, L. P., Hannivoort, R. A., Gerken, G., Hotamisligil, G. S., Trauner, M. and Canbay, A. 2012. The interaction of hepatic lipid and glucose metabolism in liver diseases. J. Hepatol. 56, 952-64.
  4. Chien, H. L., Huang, H. Y. and Chou, C. C. 2006. Transformation of isoflavone phytoestrogens during the fermentation of soymilk with lactic acid bacteria and bifidobacteria. Food. Microbiol. 23, 772-778. https://doi.org/10.1016/j.fm.2006.01.002
  5. Chun, J., Kim, G. M., Lee, K. W., Choi, I. D., Kwon, G. H., Park, J. Y., Jeong, S. J., Kim, J. S. and Kim, J. H. 2007. Conversion of isoflavone glucosides to aglycones in soymilk by fermentation with lactic acid bacteria. J. Food. Sci. 72, 39-44.
  6. Chun, J., Kim, J. S. and Kim, J. H. 2008. Enrichment of isoflavone aglycones in soymilk by fermentation with single and mixed cultures of Streptococcus infantries 12 and Weissella sp. 4. Food. Chem. 109, 278-284. https://doi.org/10.1016/j.foodchem.2007.12.024
  7. Deng, Z., Liu, Y., Liu, C., Xiang, X., Wang, J., Cheng, Z., Shah, S. V., Zhang, S., Zhang, L., Zhuang, X., Michalek, S., Grizzle, W. E. and Zhang, H. G. 2009. Immature myeloid cells induced by a high-fat diet contribute to liver inflammation. Hepatology 50, 1412-1420. https://doi.org/10.1002/hep.23148
  8. Denlinger, L. C., Fisette, P. L., Garis, K. A., Kwon, G., Vazquez-Torres, A., Simon, A. D., Nguyen, B., Proctor, R. A., Bertics, P. J. and Corbett, J. A. 1996. Regulation of inducible nitric oxide synthase expression by macrophage purinoreceptors and calcium. J. Biol. Chem. 271, 337-342. https://doi.org/10.1074/jbc.271.1.337
  9. Fried, S. K., Bunkin, D. A. and Greenberg, A. S. 1998. Omental and subcutaneous adipose tissues of obese subjects release interleukin-6: depot difference and regulation by glucocorticoid. J. Clin. Endocrinol. Metab. 83, 847-850.
  10. Gotoh, T., Yamada, K., Yin, H., Ito, A., Kataoka, T. and Dohi, K. 1998. Chemoprevention of N-Nitroso-N-methylurea-induced rat mammary carcinogenesis by soy foods or biochanin A. Jpn. J. Can. Res. 89,137-142. https://doi.org/10.1111/j.1349-7006.1998.tb00541.x
  11. Hardie, L., Trayhurn, P., Abramovich, D. and Fowler, P. 1997. Circulating leptin in women; a longitudinal study in the menstrual cycle and during pregnancy. Clin. Endocrinol. 47, 101-106. https://doi.org/10.1046/j.1365-2265.1997.2441017.x
  12. Hollister, L. E., Overall, J. E. and Snow, H. L. 1967. Relationship of obesity to serum triglyceride, cholesterol, and uric acid, and to plasma glucose levels. Am. J. Clin. Nutr. 20, 777-782. https://doi.org/10.1093/ajcn/20.7.777
  13. Hongbin, Cui. 2001. Soybean bio-functional components development and application. 241-263. Beijing: China Light Industry Press House. Beijing. China
  14. Hotamisligil, G. S. and Spiegelman, B. M. 1994. Tumor necrosis factor alpha: a key component of the obesity-diabetes link. Diabetes 43,1271-1278.
  15. Hrnciar, J., Gabor, D., Hrnciarova, M., Okapcova, J., Szentivanyi, M. and Kurray, P. 1999. Relation between cytokines (TNF-alpha, IL-1 and-6) and homocysteine in android obesity and the phenomenon of insulin resistance syndromes. Vnitr. Lek. 45, 11-16.
  16. Iritani, N., Hosomi, H., Fukuda, H., Tada, K. and Ikeda, H. 1996. Soybean protein suppresses hepatic lipogenic enzyme gene expression in Wistar fatty rats. J. Nutr. 126, 380-388. https://doi.org/10.1093/jn/126.2.380
  17. Ishimi, Y., Yoshida, M., Wakimoto, S., Wu, J., Chiba, H., Wang, X., Takeda, K. and Miyaura, C. 2002. Genistein, a soybean isoflavone, affects bone marrow lymphopoiesis and prevents bone loss in castrated male mice. Bone 31, 180-185. https://doi.org/10.1016/S8756-3282(02)00780-9
  18. Karalis, K. P., Giannogonas, P., Kodela, E., Koutmani, Y., Zoumakis, M. and Teli, T. 2009. Mechanisms of obesity and related pathology: linking immune responses to metabolic stress. FFBS. J. 276, 5747-5745.
  19. Karam, J. H., Grodsky, G. M. and Forsham, P. H. 1963. Excessive insulin response to glucose in obese subjects measured by immunochemical assay. Diabetes 12, 197-204. https://doi.org/10.2337/diab.12.3.197
  20. Kim, J. B., Sarraf, P., Wright, M., Yao, K. M., Mueller, E., Solanes, G., Lowell, B. B. and Spiegelman, B. M. 1998. Nutritional and insulin regulation of fatty acid synthetase and leptin gene expression through ADD1/SREBP1. J. Clin. Invest. 101, 1-9. https://doi.org/10.1172/JCI1411
  21. Kuo, L. C., Cheng, W. Y., Wu, R. Y., Huang, C. J. and Lee, K. T. 2006. Hydrolysis of black soybean isoflavone glycosides by Bacillus subtilis natto. Appl. Microbiol. Biotechnol. 73, 314-320. https://doi.org/10.1007/s00253-006-0474-7
  22. Lee, C. H. 2001. Fermentation technology in Korea. pp77. Korea university press. Seoul. Korea
  23. Lin, G., Wang, G., Liu, G., Yang, L. J., Chang, L. J., Lue, T. F. and Lin, C. S. 2009. Treatment of type 1 diabetes with adipose tissue-derived stem cells expressing pancreatic duodenal homeobox 1. Stem. Cells. Dev. 18, 1399-1406 https://doi.org/10.1089/scd.2009.0010
  24. Liu, D., Zhen, W., Yang, Z., Carter, J. D., Si, H. and Reynolds, K. A. 2006. Genistein acutely stimulates insulin secretion in pancreatic ${\beta}$-cells through a cAMP-dependent protein kinase pathway. Diabetes 55, 1043-1050. https://doi.org/10.2337/diabetes.55.04.06.db05-1089
  25. MacDougald, O. A. and Lane, M. D. 1995. Transcriptional regulation of gene expression during adipocyte differentiation. Annu. Rev. Biochem. 64, 345-373. https://doi.org/10.1146/annurev.bi.64.070195.002021
  26. Meli, R., Mattace, R. G., Irace, C., Simeoli, R., Di, P. A., Paciello, O., Pagano, T. B., Calignano, A., A. and Santamaria, R. 2013. High fat diet induces liver steatosis and early dysregulation of iron metabolism in rats. PLoS One 8, e66570. https://doi.org/10.1371/journal.pone.0066570
  27. Mokdad, A. H., Marks, J. S., Stroup, D. F. and Gerberding, J.L. 2005. Correction: actual causes of death in the United States. JAMA. 293, 293-294.
  28. Must, A., Spadano, J., Coakley, E. H., Field, A. E., Colditz, G. and Dietz, W. H. 1999. The disease burden associated with overweight and obesity. JAMA. 282, 1523-1529. https://doi.org/10.1001/jama.282.16.1523
  29. Nakajima, N., Nozaki, N., Ishihara, K., Ishikawa, A. and Tsuji, H. 2005. Analysis of isoflavone content in tempeh, a fermented soybean, and preparation of a new isoflavoneenriched tempeh. J. Biosci. Bioeng. 100, 685-687. https://doi.org/10.1263/jbb.100.685
  30. Nathan, C. 2006. Neutrophils and immunity: challenges and opportunities. Nat. Rev. Immunol. 6, 173-182. https://doi.org/10.1038/nri1785
  31. Park, K. Y., Jung, K. O., Rhee, S. H. and Choi, Y. H. 2003. Antimutagenic effects of doenjang (Korean fermented soypaste) and its active compounds. Mutat. Res. 523, 43-53.
  32. Peterson, T. G., Ji, G. P., Kirk, M., Coward, L., Falany, C. N. and Barnes, S. 1998. Metabolism of the isoflavones genistein and dichanin A in human breast cancer cell lines. Am. J. Clin. Nutr. 68, 1505-1511. https://doi.org/10.1093/ajcn/68.6.1505S
  33. Potter, S. M., Baum, J. A., Teng, H., Stillman, R. J., Shay, N. F. and Erdman, J. W. Jr. 1998. Soy protein and isoflavones: their effects on blood lipids and bone density in postmenopausal women. Am. J. Clin. Nutr. 68,1375-1379. https://doi.org/10.1093/ajcn/68.6.1375S
  34. Rosen, E.D. 2005. The transcriptional basis of adipocyte development. Prostaglandins. Leukot. Essent. Fatty Acids. 73, 31-34. https://doi.org/10.1016/j.plefa.2005.04.004
  35. Rosen, E D., Sarraf, P., Troy, A. E., Bradwin, G., Moore, K., Milstone, D. S., Spiegelman, B. M. and Mortensen, R. M. 1999. PPAR gamma is required for the differentiation of adipose tissue in vivo and in vitro. Mol. Cell. 4, 611-617. https://doi.org/10.1016/S1097-2765(00)80211-7
  36. Sahai, A., Malladi, P., Pan, X., Paul, R., Melin-Aldana, H. and Green, R. M. 2004. Obese and diabetic db/db mice develop marked liver fibrosis in a model of nonalcoholic steatohepatitis: role of short-form leptin receptors and osteopontin. Am. J. Physiol. Gastrointest. Liver Physiol. 287, 1035-G1043. https://doi.org/10.1152/ajpgi.00199.2004
  37. Sarraf, P., Frederich, R. C., Turner, E. M., Ma, G., Jaskowiak, N. T., Flier, J. S., Lowell, B. B., Fraker, D. L. and Alexander, H. R. 1997. Multiple cytokines and acute inflammation raise mouse leptin levels: potential role in inflammatory anorexia. J. Exp. Med. 185, 171-175. https://doi.org/10.1084/jem.185.1.171
  38. Sartipy, P. and Loskutoff, D. J. 2003. Monocyte chemoattractant protein 1 in obesity and insulin resistance. Proc. Natl. Acad. Sci. USA 100, 7265-7270. https://doi.org/10.1073/pnas.1133870100
  39. Serhan, C. N. and Savill, J. 2005. Resolution of inflammation: the beginning programs the end. Nat. Immunol. 6, 1191-1197. https://doi.org/10.1038/ni1276
  40. Sethi, J. K. and Hotamisligil, G. S. 1999. The role of TNF alpha in adipocyte metabolism. Semin. Cell. Dev. Biol. 10, 19-29. https://doi.org/10.1006/scdb.1998.0273
  41. Slavin, J. L., Karr, S. C., Hutchins, A. M. and Lampe, J. W. 1998. Influence of soybean processing, habitual diet and soy dose on urinary isoflavonoid excretion. Am. J. Clin. Nutr. 68, 1492S-1495S. https://doi.org/10.1093/ajcn/68.6.1492S
  42. Soh, J. R., Shin, D. H., Kwon, D. Y. and Cha, Y. S. 2008.Effect of Cheonggukjang supplementation upon hepatic acyl-CoA synthase, carnitine palmitoyl transferase I, acyl-CoA oxidase and uncoupling protein 2 mRNA levels in C57BL/6J mice fed with high fat diet. Genes. Nutr. 2, 365-369. https://doi.org/10.1007/s12263-007-0066-x
  43. Staiger, H., Tschritter, O., Machann, J., Thamer, C., Fritsche, A., Maerker, E., Schick, F., Haring, H. U. and Stumvoll, M. 2003. Relationship of serum adiponectin and leptin concentrations with body fat distribution in humans. Obes. Res. 11, 368-376. https://doi.org/10.1038/oby.2003.48
  44. Sugano, M., Goto, S., Yamada, Y., Yoshida, K., Hashimoto, Y., Matsuno, T. and Kimoto, M. 1999. Cholesterol-lowering activity of various undigested fraction of soybean protein in rats. J. Nutr. 12, 977-985.
  45. Teede, H. J., Dalais, F. S., Kotsopoulos, D., Liang, Y. L., Davis, S. and McGrath, B. P. 2001. Dietary soy has both beneficial and potentially adverse cardiovascular effects: a placebo- controlled study in men and postmenopausal women. J. Clin. Endocrinol. Metab. 86, 3053-3060.
  46. Todoric, J., Loffler, M., Huber, J., Bilban, M., Reimers, M., Kadl, A., Zeyda, M., Waldhausl, W. and Stulnig, T. M. 2006. Adipose tissue inflammation induced by high-fat diet in obese diabetic mice is prevented by n-3 polyunsaturated fatty acids. Diabetologia 49, 2109-2119. https://doi.org/10.1007/s00125-006-0300-x
  47. Tzang, B. S., Yang, S. F., Fu, S. G., Yang, H. C., Sun, H. L. and Chen, Y. C. 2009. Effects of flaxseed oil on cholesterol metabolism of hamsters. Food Chem. 114, 1450-1455. https://doi.org/10.1016/j.foodchem.2008.11.030
  48. Uygun, A., Kadayifci, A., Yesilova, Z., Erdil, A., Yaman, H., Saka, M., Deveci, M. S., Bagci, S., Gulsen, M., Karaeren, N. and Dagalp, K. 2000. Serum leptin levels in patients with nonalcoholic steatohepatitis. Am. J. Gastroenterol. 95, 3584-3589. https://doi.org/10.1111/j.1572-0241.2000.03297.x
  49. Weisberg, S. P., McCann, D., Desai, M., Rosenbaum, M., Leibel, F. L. and Ferrante, A. W. Jr. 2003. Obesity is associated with macrophage accumulation in adipose tissue. J. Clin. Invest. 112, 1796-1808. https://doi.org/10.1172/JCI200319246
  50. Yun, J. W., Lee, B. S., Kim, C. W. and Kim, B. H. 2007. Comparison with 3 high-fat diet for studying obesity in C57BL/6 mouse. Lab. Anim. Res. 23, 245-250.