야관문(夜關門)의 포도당 독성에 대한 세포 보호 효과

Cytoprotective Effect of Lespedeza Cuneata Extract on Glucose Toxicity

  • 최정식 (대전대학교 한의과대학 신계내과학교실) ;
  • 조충식 (대전대학교 한의과대학 신계내과학교실) ;
  • 김철중 (대전대학교 한의과대학 신계내과학교실)
  • Choi, Jeong-Sik (Dept. of Internal Medicine, College of Oriental Medicine, Daejeon University) ;
  • Cho, Chung-Sik (Dept. of Internal Medicine, College of Oriental Medicine, Daejeon University) ;
  • Kim, Cheol-Jung (Dept. of Internal Medicine, College of Oriental Medicine, Daejeon University)
  • 투고 : 2010.04.05
  • 심사 : 2010.06.16
  • 발행 : 2010.07.31

초록

Objective: Production of ROS from glucose toxicity results in injury of pancreatic $\beta$-cells in diabetes models. This study was undertaken to examine the influence of Lespedeza Cuneata extract (LCE) on cytoprotective effects on glucose toxicity, insulin secretion and gene expression in RIN-m5F cells. Methods: First, we measured LCE's antioxidant activity by DPPH free radical-scavenging activity and SOD activity. After the various concentrations of LCE were added to the RIN-m5F cells, we measured cell viability with glucose stimulation by MTT assay and glucose-stimulated insulin secretion. We analyzed gene expression with Agilent whole mouse genome 44K oligo DNA microarray and searched for related pathways in KEGG (Kyoto Encyclopedia of Genes and Genomes). Lastly we measured INS-1, INS-2, INS-R, IRS-1, IRS-2, IRS-3, GLP-1R, and GLP-2R mRNA expression by real time RT-PCR. Results: Free radical-scavenging activity, SOD activity and insulin secretion increased dependent on LCE concentration, but LCE did not show considerable cytoprotective effect on RIN-m5F cells. More than twice expressed gene was 6362 in Oligo DNA chip. In KEGG, the most related pathway was the metabolic pathway. In the insulin signaling pathway, up expressed genes were Irs1, Mapk8, Akt1, and Lipe and down expressed genes were Rhoq, Fbp2, Prkar2b, Gck, and Prkag1. In real time RT-PCR, IRS-2, and IRS-3 expression increased significantly compared to the control group on LCE $12{\mu}g/m{\ell}$ concentration and GCK expression decreased significantly compared to the control group. Conclusions: These results show that LCE encourages insulin secretion and insulin metabolism by complicated gene mechanisms. Further mechanism study and clinical study seem to be necessary about Lespedeza Cuneata.

키워드

참고문헌

  1. Robertson RP. Chronic oxidative stress as a central mechanism for glucose toxicity in pancreatic islet beta cells in diabetes. J Biol Chem. 2004;279(41):279:42351-4. https://doi.org/10.1074/jbc.R400019200
  2. Robertson RP, Harmon J, Tran PO, Tanaka Y, Takahashi H. Glucose toxicity in beta-cells: type 2 diabetes, good radicals gone bad, and the glutathione connection. Diabetes. 2003;52(3):581-7. https://doi.org/10.2337/diabetes.52.3.581
  3. Robertson RP, Harmon J, Tran PO, Poitout V. $\beta$-Cell glucose toxicity, lipotoxicity, and chronic oxidative stress in type 2 diabetes. Diabetes. 2004;53(S1):S119-24.
  4. Yoon JS, Won KC, Lee HW. Glucose Oxidation and Production of Reactive Oxygen Species (ROS) in INS-1 Cells and Rat Islet Cells Exposed to High Glucose. Diabetes. 2006;30:246-53.
  5. Wolff SP, Dean RT. Glucose autoxidation and protein modification. The potential role of 'autoxidative glycosylation' in diabetes. J Bio chem. 1987;245(1):243-50.
  6. Tiedge M, Lortz S, Drinkgern J, Lenzen S. Relation between antioxidant enzyme gene expression and antioxidative defense status of insulin-producing cells. Diabetes. 1997;46(11):1733-42. https://doi.org/10.2337/diabetes.46.11.1733
  7. Tanaka Y, Gleason CE, Tran PO, Harmon JS, Robertson RP. Prevention of glucose toxicity in HIT-T15 cells and Zucker diabetic fatty rats by antioxidants. PNAS. 1999;96:10857-62. https://doi.org/10.1073/pnas.96.19.10857
  8. Kwon MJ, Jung HS, Kim MK, Kang SH, Seo GW, Song JK et al. Diabetes. Cytoprotective Effect by Antioxidant Activity of Quercetin in INS-1 Cell Line. 2007;31(5):383-90.
  9. Ryu GR, Min DS, Rhie DJ, Yoon SH, Hahn SJ, Jo YH et al. The Inhibitory Effect of Epicatechin on IL-1$\beta$-induced iNOS Expression and NO Production in RINm5F Cell. Diabetes. 2003;27(6):456-66.
  10. Lee CW, Shin HC, Jeong JC. Effects of the Extract in Streptozotocin- induced Diabetic Rats. Korean Oriental Medical Society. 2006;27(1):91-103.
  11. Kim OG, Jeong JC. Effects of the Mori folium Extract in Streptozotocin-Induced Diabetic Rats. Journal of korean oriental internal medicine. 2006;27(4):811-21.
  12. Choi JS, Chang SK, Cho CS, Kim CJ, Han DU. Effects of Sopyung-tang Extract on Blood Glucose & Antioxidant Enzyme Activities of Streptozotocin- induced Diabetic Rats. Journal of korean oriental internal medicine. 2008;29(1): 90-103.
  13. Gong TH, Jeong JC. Effects of Ojung-hwan on Blood Glucose, Hyperlipidemia, Polyol Pathway and Antioxidative Mechanism in Ob/ob Mouse. Journal of Korean oriental medical society. 2007;28(3):57-69.
  14. Park SD, Ju WS, Go WD. Effects of Hyulbuchukeotang and its component groups on diabetes, free radical and antioxidative defense system in alloxan-induced diabetic rats. The Korea Association of Herbology. 2002; 17(1):93-111.
  15. 김창민, 신민교, 안덕균, 이경순, 강병수, 강상수 외. 중약대사전. 서울:도서출판정담. 1997:2770-3.
  16. Kim SJ, Kim DW. Antioxidative Activity of Hot Water and Ethanol Extracts of Lespedeza cuneata Seeds. Journal of the Korean Society of Post-Harvest Science & Technology of Agricultural Products. 2007;14(3):332-5.
  17. Kim MS, Min OJ, Rhyu DY. Effect of Lespedeza cuneata Extracts on Diabetes and Diabetic Nephropathy. The plant resources of society of Korea. 2008;4:83.
  18. Lee DS, Jeong GS, An RB, Bin Li, Erisa Byun, Kim YC. Search for Plant Extracts with Protective Effects of Pancreatic Beta Cell against Oxidative Stress. Kor. J. Pharmacogn. 2008;39(4):335-40.
  19. 대한당뇨병학회. 당뇨병의 진단 및 분류 권고안: 당뇨병의 분류. 임상당뇨병. 2007;8(1):31-3.
  20. Donath MY, Ehses JA, Maedler K, Shumann DM, Ellingsgaard H, Eppler E et al. Mechanisms of beta cell death in type 2 diabetes. Diabetes. 2005;54(S2):S108-13.
  21. Holman RR. Assessing the potential for $\alpha$ -glucosidase inhibitors in prediabetic states. Diabetes Res Clin Pract. 1998;40(S1):S21-5.
  22. UK Prospective Diabetes study group. Intensive blood-glucose control with sulphonylureas or insulin compared with conventional treatment and risk of complications in patients with type 2 diabetes(UKPDS 33). Lancet. 1998;352:837-53. https://doi.org/10.1016/S0140-6736(98)07019-6
  23. Kim SS, Son SM. Oxidative Stress and Cell Dysfunction in Diabetes: Role of ROS Produced by Mitochondria and NAD(P)H Oxidase. Diabetes. 2008;32:389-98.
  24. An SH, Cho CS, Kim CJ. Effects of Radix Sophora Flavescentis Extract on Insulin Secretion in RIN-m5F Cells and $\alpha$-glucosidase Inhibition. Journal of korean oriental internal medicine. 2009;30(3):481-94.
  25. Sartori A, Garay-Malpartida HM, Forni MF, Schumacher RI, Dutra F, Sogayar MC et al. Aminoacetone, a putative endogenous source of methylglyoxal, causes oxidative stress and death to insulin-producing RIN-m5F cells. Chem Res Toxicol. 2008;21:1841-50. https://doi.org/10.1021/tx8001753
  26. Jung ED, Lee JH, Jang WG, Kim JG, Kim BW, Lee IK. Microarray Analysis of Short Heterodimer Partner (SHP)- induced Changes in Gene Expression in INS-1 Cells. Diabetes. 2007;31(3):193-9.
  27. Kim JW, Joe YG, Suh KD. Poly(methyl methacrylate) hollow particles by water-in-oilin-water emulsion polymerization. Colloid Polym Sci. 1999;277:252-6. https://doi.org/10.1007/PL00013751
  28. Sakamoto Y, Higashi T. Glutathione. Japan: Scientific societies; 1989, p.5.
  29. Chung JK. Hormone Signalings Mediated by Receptor Tyrosine Kinases -Insulin model-. Korean J Endocrinology. 2000;15(3): 342-52.
  30. Son HS. Review of Insulin Signaling Network. Diabetes. 2005;29(5):383-92.
  31. Nam JH, Lee HC, Kim YE, Kwon SH, Yoon YS, Park SW et al. Glucokinase gene mutation in non-insulin- dependent diabetes mellitus(NIDDM), and secondary diabetes in Koreans. Korean J Intern Med. 1998;54(6):755-64.
  32. Xu G, Stoffers DA, Habener JF, Bonner-Weir S. Exendin-4 stimulates both beta-cell replication and neogenesis, resulting in increased beta-cell mass and improved glucose tolerance in diabetic rats. Diabetes. 1999;48:2270-6. https://doi.org/10.2337/diabetes.48.12.2270
  33. Kreymann B, Ghatei MA, Williams G, Bloom SR. Glucagon-like peptide-1(7-36): a physiological incretin in man. Lancet. 1987;2:1300-4.
  34. Komatsu R, Matsuyama T, Namba M, Watanabe N, Itoh H, Kono N et al. Glucagonostatic and insulinotropic action of glucagonlike peptide I-(7-36)-amide. Diabetes. 1989;38:902-5. https://doi.org/10.2337/diabetes.38.7.902
  35. Laurie D, Alan BT. Intestinal hormones and growth factors: Effects on the small intestine. World J Gastroenterol. 2009;15(4):385-406. https://doi.org/10.3748/wjg.15.385