A Comparative Study of Sepiae Os, Arcae Concha, Ostreae Concha and Esomeprazole in a Mouse Model of Reflux Esophagitis

Chang-Hun Song; Tae-Hyun Baek; Song, Chang-Hun; Baek, Tae-Hyun

doi:10.13048/jkm.18018

JKM > Volume 39(2); 2018 > Article

Song and Baek: A Comparative Study of Sepiae Os, Arcae Concha, Ostreae Concha and Esomeprazole in a Mouse Model of Reflux Esophagitis

Original Article

The Journal of Korean Medicine 2018; 39(2): 92-105.

Published online: June 30, 2018

DOI: https://doi.org/10.13048/jkm.18018

A Comparative Study of Sepiae Os, Arcae Concha, Ostreae Concha and Esomeprazole in a Mouse Model of Reflux Esophagitis

Chang-Hun Song, Tae-Hyun Baek

Dept. of Internal Medicine, College of Korean Medicine, Sangji University

Correspondence to: 백태현(Tae-Hyun Baek), 강원도 원주시 우산동 283번지, 상지대학교 부속한방병원 내과학교실, Tel: +82-33-741-9207, E-mail: white@sangji.ac.kr

Received May 31, 2018 Revised June 20, 2018 Accepted June 20, 2018

Abstract

Objectives

This aim of this study is to compare the reflux esophagitis improvement effects of Sepiae Os, Arcae Concha, Ostreae Concha, and Proton Pump Inhibitor(esomeprazole) through rat experiments.

Methods

NO production inhibitory effect was measured by NO production amount and iNOS mRNA expression level in cell lines. iNOS, TNF-α and p-IκB, and serotonin were compared using immunohistochemistry at the rat reflux esophagitis. Reflux esophagitis connection external form, lower esophageal sphincter, and gap were observed and an esophageal inflammatory indicator, IL-6 activity was also evaluated by immunohistochemistry.

Results

NO production and iNOS mRNA expression was showed concentration dependent decrease in cell lines treated with Sepiae OS, Arcae Concha, and Ostreae Concha at the experiments of cell lines. In the suppression of iNOS and p-IκB at the rat reflux esophagitis, Sepiae Os treat group(SOT) and Ostreae Concha treat group(OCT) were more effective. In the increase of serotonin at the rat reflux esophagitis, ACT, MT and OCT were more effective. Damage of lower esophageal sphincter, and gap between esophageal keratin and mucosa were observed less at the SOT, ACT, OCT. In the suppression of IL-6 at the rat reflux esophagitis, SOT and OCT were more effective than GE and, SOT was more effective than MT significantly.

Conclusions

The anti-inflammatory effect was the best in the SOT and lower esophageal sphincter muscle contraction was the best in the ACT at the rat reflux esophagitis. Sepiae OS was more effective than esomeprazole in the suppression of iNOS, TNF-α, and IL-6.

Keywords: Sepiae Os, Arcae Concha, Ostreae Concha, esomeprazole, reflux esophagitis

Fig. 1

Experimental design and reflux esophagitis animal model

Abbreviation. HFD, high fat diet; SO, Sepiae Os; AC, Arcae Concha; OC, Ostreae Concha.

Fig. 2

Inhibition of NO production and iNOS mRNA expression by Sepiae Os (SO)

Fig. 3

Inhibition of NO production and iNOS mRNA expression by Arcae Concha (AC)

Fig. 4

Inhibition of NO production and iNOS mRNA expression by Ostrea Concha (OC)

Fig. 5

The iNOS positive reaction in gastroesophageal junction (GEJ) with reflux esophagitis

Abbreviation. Ctrl, no reflux esophagitis elicited group; GE, reflux esophagitis elicited group; MT, medicine as esomeprazole treat group before reflux esophagitis elicitation; SOT, Sepiae Os (SO) treat group before reflux esophagitis elicitation; ACT, Arcae Concha (AC) treat group before reflux esophagitis elicitation; OCT, Ostreae Concha (OC) treat group before reflux esophagitis elicitation; Bar size, 200μm; arrow, iNOS positive reaction; LES, lower esophageal sphincter; ES, esophagus; LP, lamina propria.

Fig. 6

The TNF-α positive reaction in gastroesophageal junction (GEJ) with reflux esophagitis

Fig. 7

The p-IκB positive reaction in gastroesophageal junction (GEJ) with reflux esophagitis

Fig. 8

The Serotonin positive reaction in gastroesophageal junction (GEJ) with reflux esophagitis

Fig. 9

The improvement effect of hemorrhagic erosion in GEJ by SO, AC, OC and Esomeprazole (a photo ×4; b photo bar size 200μm)

Fig. 10

The IL-6 positive reaction in gastroesophageal junction (GEJ) with reflux esophagitis

참고문헌

1. National Professors of Bikye Internal Medicine. Bikye Internal Medicine. Seoul: Kunja Publisher;2009. 96:p. 314–318.

2. Cho YS, Choi MG, Jeong JJ. Prevalence and clinical spectrum of gastroesophageal reflux: a population-based study in Asan-si, Korea. Am J Gastroenterol. 2005; 100:747–753.

3. Lee D, Lee KJ, Kim KM, Lim SK. Prevalence of asymptomatic erosive esophagitis and factors associated with symptom presentation of erosive esophagitis. Scand J Gastroenterol. 2013; 48:906–912.

4. Kim JH, Kim JB, Kim SY, Kim SE, Kim YS, Sung HY, et al. Current Issues on Gastroesophageal Reflux Disease. Korean J Gastroenterol. 2014; 64:127–128.

5. Jung HK, Hong SJ, Jo YJ, Jeon SW, Cho YK, Lee KJ, et al. Updated guidelines 2012 for gastroenterol reflux disease. Korean J Gastroenterol. 2012; 60:4. 195–218.

6. National Professors of Herbology. Herbology. Seoul: Youngleem Publisher;1998. p. 475p. 513p. 632

7. Anderson T, Rohss K, Bredberg E, Hassan-Alim M. Pharmacokinetics and pharmacodynamics of esomeprasole, the S-isomer of omeprasole. Aliment Pharmacol Ther. 2001; 15:1563–1569.

8. Nakamura k, Ozawa Y, Furuta Y, Miyazaki H. Effects of sodiumpolyacrylate(PANa) on acute esophagitis by gastric juice in rats. Japan J Pharmacol. 1982; 32:445–456.

9. Sin GT. Steps to Internal Medicine 6 Gastrodigestive Disease. Seoul: Jungdam Publisher;2002. p. 54–55.

10. Horn J. The proton-pump inhibitors: similarities and differences. Clinther. 2000; 22:266–280.

11. Kin HJ, Leem SY, Gwak MA, Kim DJ, Byeon JS. Effects of Yijintang-gamibang on Reflux Esophagitis Induced by Pylorus and Forestomach Ligition in Rat. J of Internal Korean Medicine. 2010; 31:1. 128–141.

12. Heo JI. Protective Effects of Yijin-tang-gamib-molyeo Aqueous Extracts on Reflux Esophagitis. PhD Dissertation. Daegu: Daegu Haany University;2011.

13. Dent J. Pharmacology of esomeprazole and comparisons with omeprazole. Aliment Pharmacol Ther. 2003; 17:suppl 1. 5–9.

14. Pharmscore. Available at: http://www.pharmscore.com/news/digestive_pharms/treatarticles/esomepr1.htmlAccessed June 21, 2016.

15. Fass R, Dickman R. Non erosive reflux disease. GI Motility online. 2006; 10.1038/gimo42

16. Fauci. Harrison’s Internal Medicine. Seoul: MIP Publisher;2010. p. 2234

17. Park K, Gwak MA, Kim DJ, Byeon JS. Protective Effects of Yijin-tang-gamibang Aqueous Extracts on Reflux Esophagitis Mediated by Antioxidant Defense Systems. Korean J of Oriental Physiology and Pathology. 2010; 24:3. 416–425.

18. Jang MW, Leem SW. Experimental Study for Effect of Banhasasim-tang on Mice with Reflux Esophagitis. J of Internal Korean Medicine. 2013; 34:4. 362–374.

19. Kim MY, Sin YO, Lee JY, Lee AR, Sin SH, Kwon OJ, et al. Improving Effect of a Combined Extract of Rhei Rhizoma and Glycyrrhizae Rhizoma through Anti-oxidative Stress in Reflux Esophagitis rats. Korea J of Herbology. 2015; 30:4. 37–44.

20. Kim DJ, No SS. Effect on Acute reflux Esophagitis by Evodiae Fructus Aquous Extract. Korea J of Herbology. 2012; 27:1. 51–58.

21. Lee YJ, Park JH, No SS. Effects on Rats with Reflux Esophagitis Treated with Lonicerae Flos Extract. Korean J of Oriental Physiology and Pathology. 2010; 24:6. 970–975.

22. Chinese Medicine Dictionary Publish Members. Chinese Medicine Dictionary. Seoul: Jungdam Publisher;1997. p. 1317–1320. p. 3123–3124. p. 4722–4726.

23. Lee NK, Yun HI, Park SC, Park JI, Jo MH. Effects of heat-treated alumen, halloysitum rubrum and os sepiae in experimentally induced stomach ulcer in rats. J of Applied Pharmacology. 1997; 5:246–252.

24. Kim TG, Lee YH, Choi YS, Byeon JS, Park SD. The Effects of ShogunJungtang-ga-younggol. morea on Gastric Ulcer. J of Internal Korean Medicine. 2001; 22:1. 13–20.

25. Patk SM, Jo YG, Kee JR, Lee YC, Kim HJ, Kwon YK, et al. Inhibitory Effect of Oyster Conchioloin on Pro-inflammatory Mediator in Lipopolysaccharide;Activated Raw 264.7 Cells. Korean J of Oriental Physiology and Pathology. 2008; 22:4. 878–883.

26. Lee YT, Choi BT, Choi Yh, Kang KH. Development of Health Assistances for Anti Stress used with Ostreae Concha. Korean J of Oriental Physiology and Pathology. 2006; 20:6. 1604–1611.

27. Itzkowitz SH, Yio X. Inflammation and cancer IV. Colorectal cancer in inflammatory bowel disease: the role of inflammation. Am J Physiol Gastrointest Liver Physiol. 2004; 287:1. G7–17.

28. Stamler JS, Singel DJ, Loscalzo J. Biochemistry of nitric oxide and its redox activated forms. Science. 1992; 258:1898–1902.

29. De May JG, Vanhoutte PM. Rolo of the intima in cholinergic and purinergic relaxation of isolated canin femoral arteries. Journal of Physiology. 1981; 316:347–355.

30. Furchgott RF, Zawadzki JV. The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature. 1980; 288:373–376.

31. Brennan FM, Maini RN, Feldmann M. Cytokine expression in chronic inflammatory disease. Br Med Bull. 1995; 51:2. 368–84.

32. Xie QW, Kashiwabara Y, Nathan C. Role of transcription factor NF-kappa B/Rel in induction of nitric oxide synthase. J Biol Chem. 18. 269:7. 4705–8. 1994.

33. Barnes PJ, Karin MN. Nuclear factor-kappaB: a pivotal transcription factor in chronic inflammatory diseases. Engl J Med. 10. 336:15. 1066–71. 1997.

34. Baeuerle PA, Baltimore D. NF-κB - Ten years after. Cell. 1996; 87:13–20.

35. Seo WG, Pae HO, Oh GS, Chai KY, Yun YG, Kwon TO, et al. Inhibitory effect of ethyl acetate fraction from Cudrania tricuspidata on the expression of nitric oxide synthase gene in RAW 264.7 macrophages stimulated with interferon-and lipopolysaccharide. Gen Pharmacol. 2000; 35:21–28.

36. Seo WG, Pae HO, Oh GS, Chai KY, Kwon TO, Yun YG, et al. Inhibitory effects of methanol extracts of Cyrerus rotundus rhizomes on nitric oxide and superoxide production by murine macrophages cell line, RAW 264.7 cells. J Ethnopharmacol. 2001; 76:59–64.

37. Lee BG, Kim SH, Zee OP, Lee KR, Lee HY, Han JW, et al. Suppression of inducible nitric oxide synthase expression in RAW 264.7 macrophagesby two-carboline alkaloids extractes from Melia azedarach. Eur J Pharmacol. 2000; 406:301–309.

38. Lee DA, Lee JR, Kim YW, Kwon YK, Byun SH, Sin SW, et al. Inhibition of Lipopolysaccharide-Inducible Nitric Oxide Synthase, TNF-α, IL-1β and COX-2 Expression by Flower and Whole Plant of Lonicera japonica. Korean J of Oriental Physiology and Pathology. 2005; 19:2. 481–489.

39. Byeon SH, Yang JH, Kim SC. Inhibitory effect of Scrophulariae Radix extract on TNF-a, IL-1β, IL-6 and Nitric Oxide production in Lipopolysaccharide - activated Raw 264.7 cells. Korea J of Herbology. 2005; 20:2. 7–16.

40. Racke K, Riemann A, Schworer H, Kilbinger H. Regulation of 5-HT release from enterochromaffin cell. Behav Brain Res. 1996; 73:83–7.

41. Morrow JD, Roberts LJ. Lipid-derived autocoides, eicosanoids and platelets-activating factor. Hardman JG, Limberd LE, editors. Goodman and Gilman’s The Pharmacological Basis of Therapeutics. USA: McGraw Hill and Co;2001. p. 673

42. Saegusa Y, Takeda H, Muto S, Oridate N, Nakagawa K, Sadakane C, et al. Decreased motility of the lower esophageal sphincter in a rat model of gastroesophageal reflux disease may be mediated by reductions of serotonin and acetylcholine signaling. Biol Pharm Bull. 2011; 34:5. 704–11.

43. Tobey NA, Carson JL, Alkiek RA, Orlando RC. Dilated intercellular spaces: a morphlogical feature of acid reflux – damaged human esophageal epithelium. Gastroenterology. 1996; 111:5. 1200–5.

44. De Hertogh G, Ectors N, Van Eyken P, Geboes K. Review article: the nature of oesophageal injury in gastro-oesophageal reflux disease. Aliment Pharmacol Ther. 2006; 24:Suppl2. 17–26.

45. Ardizzone S, Bianchi Porro G. Biologic therapy for inflammatory bowel disease. Drugs. 2005; 65:16. 2253–86.

46. Stamler JS, Singel DJ, Loscalzo J. Biochemistry of nitric oxide and its redox activated forms. Science. 1992; 258:1898–1902.

47. De May JG, Vanhoutte PM. Rolo of the intima in cholinergic and purinergic relaxation of isolated canin femoral arteries. Journal of Physiology. 1981; 316:347–355.

48. Furchgott RF, Zawadzki JV. The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature. 1980; 288:373–376.

49. Han GJ, Leem JT, Kim JS, Lee JH. Analysis of the Existing Guidelines and Clinical Trials for the Development of the Guidelines of Clinical Trials with Herbal Medicinal Products for Gastroesophageal Reflux Disease(GERD). J of Internal Korean Medicine. 2016; 37:1. 90–108.