생약조성물(MH-30)의 면역조혈계 및 재생조직 방사선 손상에 대한 방호 효과

국문 초록 (Abstract)

본 연구팀에서는 방사선에 의한 면역조혈계 및 위장관계 손상에 대한 방호를 위하여 당귀, 천궁, 작약 혼합물의 열수추출물로부터 그 다당체 함량을 강화한 생약조성물 Hemo HIM을 개발한 바 있다. 본 연구에서는 열수 추출물에 기반을 두어 제조된 HemoHIM에 비하여 지용성 폴리페놀 성분을 강화함으로써 더 뛰어난 생리활성을 갖는 생약조성물을 개발하고자 30% 에탄올 추출과 열수 추출을 함께 시행하여 얻은 추출물을 기반으로 생약조성물 MH-30을 제조하였다. MH-30과 HemoHIM의 에탄올 분획 내의 성분을 HPLC로 비교 분석한 결과 수용성 성분에는 큰 차이가 없었으나 여러가지 지용성 성분이 MH-30에서 크게 증가하는 것을 확인하였으며, 특히 decursin의 함량은 8.7배로 크게 증가하였다. 다음으로 MH-30의 시험관 내 항산화 및 면역세포 활성화 효과와 방사선 조사 마우스에서 면역조혈계 및 재생조직의 방호 효과를 HemoHIM과 비교하여 관찰하였다. 항산화 활성을 비교하기 위한 시험관 내 hydroxyl radical 및 superoxide anion 소거 활성 평가에서 MH-30이 HemoHIM 보다 높은 항산화 활성을 보였다. 림프구 증식능을 이용한 시험관 내 면역세포 활성화 시험에서는 MH-30과 Hemo HIM은 거의 비슷한 활성을 나타내었다. 방사선 조사 마우스에서 MH-30은 내재성 비장 조혈세포 집락수를 증가시키고 골수조직 내 세포사멸을 줄였으며, 위장관 재생조직인 소장움의 생존율을 증가시키는 등 방사선에 의한 면역조혈계와 재생조직 손상을 방호하는 효과를 보여주었다. 또한, 종합적인 방사선 방호 효과를 평가하는 지표인 방사선 조사 마우스의 30일 생존율도 MH-30 투여로 유의하게 증가함을 확인하였다. 특히 상기한 모든 마우스 실험에서 MH-30은 Hemo HIM보다 더 뛰어난 방호 효과가 있음을 관찰할 수 있었다. 이상의 결과들은 새롭게 개발한 생약조성물 MH-30이 면역조혈계와 재생조직의 방사선 손상을 줄여주는 효과가 있으며 기존에 개발된 HemoHIM에 비해 더 뛰어난 활성을 갖고 있음을 보여주었다. 따라서 MH-30은 방사선 사고 또는 암환자의 방사선치료 시 발생할 수 있는 면역조혈계 및 재생조직의 손상을 경감시킬 수 있는 방사선 방호 물질로서 유용하게 활용될 수 있을 것으로 생각한다.

다국어 초록 (Multilingual Abstract)

We previously developed an herbal composition (HemoHIM) based on the water extracts of Angelica gigas radix, Cnidium officinale rhizoma, and Paeonia japonica radix to protect and recover hematopoietic and intestinal tissues against radiation injuries. In this study, to develop a composition with improved activities based on enhanced fat-soluble polyphenol contents, we prepared a new herbal composition, MH-30, from the above three herbs by 30% ethanol extraction and hot water extraction. HPLC analysis of the ethanol fractions of MH-30 and HemoHIM revealed that MH-30 had higher contents of many fat-soluble polyphenol compounds than HemoHIM (8.7-fold increase for decursin), whereas contents of water-soluble polyphenol compounds showed little differences between the two compositions. Then, we evaluated MH-30 and HemoHIM for their in vitro antioxidant and immune cell-stimulating activities as well as in vivo protective effects against radiation injuries in hematopoietic and self-renewal tissues. In antioxidant activity assays, MH-30 showed higher hydroxyl radical scavenging activity than HemoHIM (1.4- to 1.9-fold for compositions and 2.3- to 4.5-fold for ethanol fractions). On the other hand, MH-30 and HemoHIM exhibited similar immune cell-stimulating activities as measured by in vitro lymphocyte proliferation. MH-30 increased endogenous spleen colony formation, decreased bone marrow cell apoptosis, and enhanced survival of intestinal crypts in irradiated mice, demonstrating effective protection of MH-30 against radiation-induced injuries in hematopoietic and self-renewal tissues. The 30-day survival rate of lethally irradiated mice, a comprehensive index for radioprotective efficacy, was also elevated by MH-30. Noticeably, MH-30 showed higher protective effects than HemoHIM in all mouse experiments. These results demonstrate that MH-30 can protect hematopoietic and self-renewal tissues against radiation injuries more effectively than HemoHIM. Therefore, MH-30 can be a good candidate to reduce radiation injuries in hematopoietic and self-renewal tissues incurred by radiation accidents or cancer radiation therapy.

목차 (Table of Contents)

• ABSTRACT
• 서론
• 재료 및 방법
• 결과
• 고찰
• 요약
• REFERENCES

참고문헌 (Reference)

1 조성기, "방사선에 대한 생약복합조성물(HemoHIM)의 재생조직 및 면역계 방호·회복촉진 효과" 한국식품영양과학회 34 (34): 805-813, 2005

2 조성기, "면역조혈계 및 재생조직의 방사선 손상에 대한 생약복합물(HIM-I)의 방호 효과" 한국식품영양과학회 34 (34): 605-612, 2005

3 Singh VK., "γ-Tocotrienol as a promising countermeasure for acute radiation syndrome: Current Status" 37 : E663-, 2016

4 MacVittie TJ., "Therapeutic use of recombinant human G-CSF(rhG-CSF)in a canine model of sublethal and lethal whole-body irradiation" 57 : 723-736, 1990

5 Kim SH., "The radioprotective effects of bu-zhongyi-qi-tang : a prescription of traditional Chinese medicine" 30 : 127-137, 2002

6 Hendry JH., "The clonogen content of murine intestinal crypts : dependence on radiation dose used in its determination" 132 : 115-119, 1992

7 Ohta S., "Studies on chemical protectors against radiation. ⅩⅩⅩ. Radioprotective substances of Cnidii rhizoma" 110 : 746-754, 1990

8 Rades D., "Serious adverse effects of amifostine during radiotherapy in head and neck cancer patients" 70 : 261-264, 2004

9 Neta R, "Role of cytokines in radioprotection" 39 : 261-266, 1988

10 Park HR., "Restoration of the immune functions in aged mice by supplementation with a new herbal composition, HemoHIM" 22 : 36-42, 2008

11 Schwartz GN., "Recovery of hematopoietic colony-forming cells in irradiated mice pretreated with interleukin 1(IL-1)" 16 : 752-757, 1988

12 Gutteridge JM, "Reactivity of hydroxyl and hydroxyllike radicals discriminated by release of thiobarbituric acidreactive material from deoxy sugars, nucleosides and benzoate" 15 : 761-767, 1984

13 Milas L., "Radioprotectors in tumor radiotherapy : factors and settings determining therapeutic ratio" 39 : 179-187, 1988

14 Lee SE., "Radioprotective effects of two traditional Chinese medicine prescriptions : si-wu-tang and si-jun-zi-tang" 27 : 387-396, 1999

15 Nair GG., "Radioprotective effects of gallic acid in mice" 2013 : 953079-, 2013

16 Jiang S., "Radioprotective effects of Sipunculus nudus L. polysaccharide combined with WR-2721, rhIL-11 and rhG-CSF on radiation-injured mice" 56 : 515-522, 2015

17 Hsu HY., "Radioprotective effect of Ganoderma lucidum(Leyss. ex. Fr. )Karst after X-ray irradiation in mice" 18 : 61-69, 1990

18 Jothy SL., "Radioprotective activity of Polyalthia longifolia standardized extract against X-ray radiation injury in mice" 32 : 150-161, 2016

19 Miyanomae T., "Radioprotection of hemopoiesis conferred by Acanthopanax senticosus Harms(Shigoka)administered before or after irradiation" 16 : 801-806, 1988

20 Pamujula S., "Radioprotection in mice following oral delivery of amifostine nanoparticles" 81 : 251-257, 2005

21 Hall EJ., "Radiobiology for the radiologist. 3rd ed" J.B. Lippincott Company 365-376, 1988

22 Milas L., "Protective effects of S-2-(3-aminopropylamino)ethylphosphorothioic acid against radiation damage of normal tissues and a fibrosarcoma in mice" 42 : 1888-1987, 1982

23 Park HR., "Protective effects of HemoHIM on immune and hematopoietic systems against γ-irradiation" 28 : 245-251, 2014

24 김성호, "Protective Effect of an Herbal Preparation (HemoHIM) on Radiation-Induced Intestinal Injury in Mice" 한국식품영양과학회 12 (12): 1353-1358, 2009

25 Travis EL., "Protection of mouse bone marrow by WR-2721 after fractionated irradiation" 15 : 377-382, 1988

26 Hsu HY., "Protection of mouse bone marrow by Si-WU-Tang against whole body irradiation" 52 : 113-117, 1996

27 Kim JJ., "Preventative effect of an herbal preparation(HemoHIM)on development of airway inflammation in mice via modulation of Th1/2 cells differentiation" 8 : e68552-, 2013

28 Withers HR., "Microcolony survival assay for cells of mouse intestinal mucosa exposed to radiation" 17 : 261-267, 1970

29 Singh VK., "Medical Countermeasures for radiation exposure and related injuries : Characterization of medicines, FDA-approval status and inclusion into the strategic national stockpile" 108 : 607-630, 2015

30 Preeti Verma, "Management of Radiation Injuries by Panax ginseng Extract" 고려인삼학회 35 (35): 261-271, 2011

31 Hisha H., "Isolation and identification of hematopoietic stem cell-stimulating substances from Kampo(Japanese herbal)medicine, Juzen-taiho-to" 90 : 1022-1030, 1997

32 Neta R., "Interleukin 1 is a radioprotector" 136 : 2483-2485, 1986

33 Milas L., "In vivo radioprotective activities of diethyldithiocarbamate(DDC)" 10 : 2335-2343, 1984

34 Zneg XL., "Immunological and hematopoietic effect of Codonopsis pilosula on cancer patients during radiotherapy" 12 : 607-608, 1992

35 Park HR., "HemoHIM ameliorates the persistent down-regulation of Th1-like immune responses in fractionated γ-irradiated mice by modulating the IL-12p70-STAT4 signaling pathway" 177 : 676-684, 2013

36 박혜란, "HemoHIM Enhances the Therapeutic Efficacy of Ionizing Radiation Treatment in Tumor-Bearing Mice" 한국식품영양과학회 13 (13): 47-53, 2010

37 Halliwell B., "Free radicals in biology and medicine. 3rd ed" Oxford University Press 604-607, 1999

38 Chaudhary S., "Evaluation of antioxidant and anticancer activity of extract and fractions of Nardostachys jatamansi DC in breast carcinoma" 15 : 50-, 2015

39 Park HR., "Enhanced antitumor efficacy of cisplatin in combination with HemoHIM in tumor-bearing mice" 9 : 85-, 2009

40 Duke RC., "Endogenous endonuclease-induced DNA fragmentation : an early event in cell-mediated cytolysis" 80 : 6361-6315, 1983

41 Quan HX., "Effects of radix Astragali on hemopoiesis in irradiated mice" 19 : 741-743, 1994

42 Hsu HY., "Effects of kuei-pi-tang on cellular immunocompetence of gamma-irradiated mice" 21 : 151-158, 1993

43 Asghar N., "Compositional difference in antioxidant and antibacterial activity of all parts of the Carica papaya using different solvents" 10 : 5-, 2016

44 Satoh S., "Case of malignant melanoma of the external genitalia responding satisfactorily to a combination of local injection of OK-432and chemotherapy" 9 : 140-145, 1982

45 Bogo V., "Behavioral toxicity and efficacy of WR-2721 as a radioprotectant" 104 : 182-190, 1985

46 Ghosh SP., "Amelioration of radiation-induced hematopoietic and gastrointestinal damage by Ex-RAD(R)in mice" 53 : 526-536, 2012

47 Mei QB., "Advances in the pharmacological studies of radix Angelica sinensis(Oliv)Diels(Chinese Danggui)" 104 : 776-781, 1991

48 Nicoletti I., "A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry" 139 : 271-279, 1991

49 Peskin AV., "A microtiter plate assay for superoxide dismutase using a water-soluble tetrazolium salt(WST-1)" 293 : 157-166, 2000