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녹용대보탕 열수 추출물의 실험적으로 유발된 랫트 아급성 출혈성 빈혈에 대한 효과

Ameliorating Effects of Nokyongdaebo-tang on Experimental Subacute Hemorrhagic Anemia in Rats

大韓韓方婦人科學會誌 = The Journal of Korean obstetrics & gynecology, v.30 no.3, 2017년, pp.1 - 19  

김정아 (부산대학교 한방병원 여성의학과) ,  김동철 (대구한의대학교 한의과대학 부인과교실)

초록
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목 적: 본 연구는 녹용대보탕이 안와정맥총 방혈로 유발시킨 랫트의 아급성 출혈성 빈혈 모델에 대해 항빈혈 및 조혈촉진 효과를 확인하였다. 방 법: 본 실험에서는 안와 정맥총에서 혈액을 채혈하여 아급성 출혈성 빈혈을 유발시켰으며, 녹용대보탕 추출물을 경구투여하고, 체중 및 조혈 관련 장기(대퇴골 골수, 비장 및 간) 중량, 적혈구 관련 혈액학적 지표의 변화, 혈액 도말 표본의 세포학적 변화, 조혈관련 장기의 조직병리학적 변화와 함께 조혈장기 내 혈구형성 줄기세포의 수적 변화를 면역조직화학적으로 관찰하였다. 또한 철분 보충제의 대표적 부작용인 소화 장애, 특히 변비 유발 여부를 확인하기 위해 탄분을 이용해 위장관 운동성을 평가하였으며, 결장내 분변 표면의 점막 두께와 결장점막 내 점액생산세포 및 결장 점막 두께 변화를 조직병리학적으로 평가하였다. 결 과: 본 실험에서 유발된 아급성 출혈성 빈혈 소견이 모든 세 용량의 녹용대보탕 추출물의 용량 의존적으로 현저히 억제되었다. 또한 세 용량의 녹용대보탕 투여군에서는 정상 대조군 및 아급성 출혈성 빈혈 대조군에 비해 위장관 운동성 변화 및 결장 및 결장내 분변의 점액성분의 유의미한 변화는 없었다. 결 론: 이상의 결과에서, 녹용대보탕의 경구투여는 아급성 출혈성 빈혈 유발 랫트에서 단순 철분 보충제의 부작용인 소화관 장애 없이, 조혈관련 장기들에서 혈구형성줄기세포의 증식 촉진을 통해 투여 용량 의존적으로 철 결핍성 빈혈의 증상을 현저히 완화시키는 것으로 관찰되었다. 따라서 녹용대보탕은 새로운 개념의 조혈촉진 빈혈치료제로서의 개발 가치가 충분할 것으로 판단된다.

Abstract AI-Helper 아이콘AI-Helper

Objectives: The object of this study is to observe the possible ameliorating effects of Nokyongdaebo-tang (NYDBT) on the experimental subacute hemorrhagic anemia (SHA) in rats. Methods: In the present study, SHA in rats was induced by exsanguinations from orbital plexus, and ameliorating effects of ...

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제안 방법

  • Items for hematology measurement are listed below and the detecting methods and unit measured are presented as follow items, respectively. All hematological measurements were conducted in Veterinary Teaching Hospital, College of Veterinary Medicine, Kyungpook National University (Daegu, Korea) using automated hematology cell counter (MS9-5V; Melet Schloesing Lab., Paris, France).
  • Changes of body weight were checked at once a day from 1 day before initial exsanguinations and test material throughout all periods of experiment using an automatic electronic balance (Precisa Instrument, Zuerich, Switzland). To reduce the differences of individuals, the body weight gains after 7 days of administrations were calculated as follow Equation [1].
  • Moreover, FM 5 ml/kg also induced noticeable and significant decreases of mucous components in the colon and remnant feces in situ under alcian blue staining as direct evidences that they induced constipation. However, no meaningful changes on the charcoal transfers, mucous components in the colonic mucosa and also in remnant feces were demonstrated in all three different dosages of NYDBT as compared to those of intact vehicle and SHA control in this experiment. These are direct evidences that NYDBT has potent favorable anti-anemic effects on SHA through proliferating effects on hematopoietic stem cells with less digestive discomfort, constipations, one of major problems in iron supplements4).
  • , Armonk, NY, USA). In addition, the percent changes between intact vehicle and SHA control rats were calculated to observe the severities of SHA induced by continuous exsanguinations in this study, and the percent changes as compared with SHA control and test material treated rats were also calculated to help the understanding of the anti-anemic effects of test substances as follow Equation[4] and [5], according to previous method described by Kang et al22), respectively.
  • The cells taken by over 20% of immunoreactivities, the density of CD34 and CD45, were regarded as positive. The mean numbers of CD34 and CD45-immunoreactive cells dispersed in the liver, spleen and femur bone marrow (mm2) were counted using a computer-assisted image analysis program, respectively. The histopathologist was blinded to the group distribution when performing the analysis.
  • Therefore, in this study, we examined the ameliorating effects of NYDBT on experimental subacute hemorrhagic anemia in rats and compared with commercial iron supplements, FerromaxTM (7.14% ferric hydroxide polymaltose complex; FM, Hanmi Pharm. Co. Ltd., Seoul, Korea).

대상 데이터

  • Total sixty healthy female SPF/VAF Crl : CD1 [Sprague-Dawley] rats (6 weeks old upon receipt; Orient Bio, Seungnam, Korea; Body weight ranged in 120~150 g upon receipt), were used after acclimatization for 8 days. Animals were assigned four to five per polycarbonate cage in a temperature (20~25℃) and humidity (50~55%) controlled room.

데이터처리

  • Variance homogeneity was examined using the Levene test21). If the Levene test signified no significant deviations from variance homogeneity, the obtain data was analyzed by one way ANOVA test followed by least-significant differences multi-comparison (LSD) test to determine which pairs of group comparison were significantly different. In case of significant deviations from variance homogeneity were observed at Levene test, a non-parametric comparison test, Kruskal-Wallis H test was conducted.

이론/모형

  • Multiple comparison tests for all dose groups were treated. Variance homogeneity was examined using the Levene test21). If the Levene test signified no significant deviations from variance homogeneity, the obtain data was analyzed by one way ANOVA test followed by least-significant differences multi-comparison (LSD) test to determine which pairs of group comparison were significantly different.
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참고문헌 (27)

  1. Shin TF. Steps to Internal Medicine 1. Seoul:Jungdam. 2005:29-66. 

  2. HaBelmann J, Dornbusch J. A rare cause of a chronic bleeding in the upper gastrointestinal tract. Chirurg. 2013;84(4):322-4. 

  3. Bernardi LA, et al. The association between subjective assessment of menstrual bleeding and measures of iron deficiency anemia in premenopausal African-American women: a cross-sectional study. BMC Womens Health. 2016;16(1):50. 

  4. Geisser P. Safety and efficacy of iron(III)-hydroxide polymaltose complex/a review of over 25 years experience. Arzneimittelforschung. 2007;57(6A):439-52. 

  5. Feltrin C, et al. Effect of soluble fiber pectin on growth and intestinal iron absorption in rats during recovery from iron deficiency anemia. Biol Trace Elem Res. 2009;129(1-3):221-8. 

  6. Shin CH. Catechetical Basics of Korean medicine. Seoul:Seongbosa. 1988:240. 

  7. Kim WH. Byeonjeungronchi of internal organs(臟腑辨證論治). Seoul:Seongbosa. 1985:60-2. 

  8. Lee C. Uihagimmun(醫學入門). Seoul:Bubin. 2009:1900. 

  9. Cheong MH, Jung IC, Lee SR. The neuroprotective effects of Nokyongdaebo-tang (Lurongdabutang) treatment in pathological Alzheimer's disease model of neural tissues. J Oriental Neuropsychiatry. 2009;20(2):1-17. 

  10. Lee SW, et al. Preventive effects of Nokyongdaebotang on hyperlipidemia rats. Kor J Oriental Preventive Medical Society. 2003;7(2):107-19. 

  11. Lee KA, Chung HY. Biological activities of a Korean traditional prescription, Nogyongdaebotang. J Korean Soc Food Sci Nutr. 2004;33(1):28-33. 

  12. Bae NY, Ahn TW. The anti-oxidative effect of oral administration of NYD (Nocyongdaebo-tang) in oxidized rats induced by AAPH. J Sasang Constitutional Medicine. 2007;19(2):155-69. 

  13. Lee SY, Ahn TW. Anti-aging effect of Tae-Eumin's Nocyongdaebo-tang (NYD) in aged rats. J Sasang Constitutional Medicine. 2008;20(2):58-71. 

  14. Pulina MO, et al. Effect of lactoferrin on consequences of acute experimental hemorrhagic anemia in rats. Bull Exp Biol Med. 2010;149(2):219-22. 

  15. Sagar L, Sehgal R, Ojha S. Evaluation of antimotility effect of Lantana camara L. var. acuelata constituents on neostigmine induced gastrointestinal transit in mice. BMC Complement Altern Med. 2005;5:18. 

  16. Choi JS, et al. Synergistic effect of fermented rice extracts on the probiotic and laxative properties of yoghurt in rats with loperamide-induced constipation. Evid Based Complement Alternat Med. 2014;2014:878503. 

  17. Song MY. Effect of low molecular weight fucoidan on genotoxicity and chronic cisplatin-induced delayed gastrointestinal motility. Seoul:Konkuk University. 2013:36-8. 

  18. Price EA, et al. Anemia in older persons: etiology and evaluation. Blood Cells Mol Dis. 2011;46(2):159-65. 

  19. Wu D, et al. Traditional Chinese formula, lubricating gut pill, stimulates cAMP-dependent Cl(-) secretion across rat distal colonic mucosa. J Ethnopharmacol. 2010;134(2):406-13. 

  20. Tang X, et al. Antigen-retrieval procedure for bromodeoxyuridine immunolabeling with concurrent labeling of nuclear DNA and antigens damaged by HCl pretreatment. J Neurosci. 2007;27(22):5837-44. 

  21. Levene A. Pathological factors influencing excision of tumours in the head and neck. Part I. Clin Otolaryngol Allied Sci. 1981;6(2):145-51. 

  22. Kang SJ, et al. Fermentation with Aquilariae Lignum enhances the anti-diabetic activity of green tea in type II diabetic db/db mouse. Nutrients. 2014;6(9):3536-71. 

  23. Moghadam AM, et al. Relationship between blood donors' iron status and their age, body mass index and donation frequency. Sao Paulo Med J. 2013;131(6):377-83. 

  24. Scholbach J, et al. Comparison of hematopoietic stem cells derived from fresh and cryopreserved whole cord blood in the generation of humanized mice. PLoS One. 2012;7(10):e46772. 

  25. Melikian AL, et al. Hereditary spherocytic hemolytic anemia in an adult with the formation of ectopic foci of extramedullary hemopoiesis in the chest. Ter Arkh. 2010;82(7):72-5. 

  26. Baranski GM, et al. ${\beta}$ -blockade protection of bone marrow following trauma: the role of G-CSF. J Surg Res. 2011;170(2):325-31. 

  27. Wintola OA, Sunmonu TO, Afolayan AJ. The effect of Aloe ferox Mill. in the treatment of loperamide-induced constipation in Wistar rats. BMC Gastroenterol. 2010;10:95. 

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