포도당 처리로 유도된 뇌신경세포 독성에 대한 눈개승마 추출물의 보호효과

국문 초록 (Abstract)

본 연구에서는 눈개승마(Aruncus dioicus var. kamtschaticus)의 in vitro 산화방지활성 및 고당(intensive glucose)과 과산화수소(H₂O₂)로 야기된 산화적 스트레스로부터의 뇌신경세포 손상에 대한 보호효과와 더불어 알파글루코사이드가수분해효소 및 아세틸콜린에스터가수분해효소 억제효과를 확인하였으며 또한 HPLC를 이용하여 주요 물질을 분석하였다. 눈개승마 아세트산에틸 분획물(ethylacetate fraction from Aruncus dioicus var. kamtschaticus: EFAD)은 매우 우수한 총 페놀화합물 함량(430.08 mg GAE/g)과 총 플라보노이드 함량(511.72 mg RE/g)을 나타냈으며, 높은 ABTS 라디칼 소거 활성과 과산화지방질생성물의 억제력을 확인하였다. 또한 고당(intensive glucose)과 과산화수소(H₂O₂)로 야기된 뇌신경세포에서의 산화적 스트레스 및 이로 인한 뇌신경세포 사멸을 측정한 결과, 눈개승마 아세트산에틸 분획물(EFAD)의 유의적인 뇌신경세포 보호효과를 확인할 수 있었다. 추가적으로 다당류 분해효소인 알파글루코사이드가수분해효소 억제효과 및 아세틸콜린 분해 효소인 아세틸콜린에스터가수분해효소 억제효과를 살펴봄으로써 혈당 상승 억제효과와 뇌신경전달물질의 세포 내 유지 효과를 확인하였다. 마지막으로 눈개승마 아세트산에틸 분획물(EFAD)의 주요 페놀성 물질을 확인하기 위해 HPLC분석을 한 결과 카페산으로 추정되었다. 본 연구 결과들을 고려할 때, 눈개승마는 고혈당 지연 또는 개선을 통한 고혈당 예방 소재로서의 가능성 뿐만 아니라 이로 인해 야기되는 산화적 스트레스로부터 뇌신경 세포를 보호함으로써 고혈당에 의한 대사성 뇌신경질환 예방 소재로도 활용 가능성이 있을 것이라 판단된다.

다국어 초록 (Multilingual Abstract)

To assess the physiological effects of Aruncus dioicus var. kamtschaticus extract on cytoxicity of a neuronal cell line, antioxidant activity, and neuroprotection against intensive glucose-induced oxidative stress were quantitated. Compared to the other fractions, the ethyl acetate fraction of Aruncus dioicus var. kamtschaticus (EFAD) showed the highest total phenolics and flavonoids. The 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assay and malondialdehyde inhibitory effect test confirmed the superior antioxidant activity of EFAD. Moreover, EFAD also decreased the intracellular ROS level and suppressed neuronal cell death against intensive glucose- or H₂O₂-induced oxidative stress. Additionally, assessment of α-glucosidase and acetylcholinesterase inhibitory activities revealed that EFAD was an effective inhibitor of α-glucosidase and acetylcholinesterase. Finally, high-performance liquid chromatography analysis identified caffeic acid as the main ingredient of EFAD. Overall, these results suggest that the EFAD is a good natural source of biological compounds that counteract hyperglycemic neuronal defects.

목차 (Table of Contents)

• Abstract
• 서론
• 재료 및 방법
• 결과 및 고찰
• 요약
• References

참고문헌 (Reference)

1 정창호, "코코아 폴리페놀 성분의 in vitro 항산화 활성" 한국식품저장유통학회 17 (17): 100-106, 2010

2 정희록, "생마늘 추출 숙성물의 항산화 및 아세틸콜린에스터라아제 저해 효과" 농업생명과학연구원 45 (45): 113-120, 2011

3 김미선, "눈개승마(Aruncus dioicus var. kamtschaticus Hara) 용매 추출물의 항산화 및 항균활성" 한국식품영양과학회 40 (40): 47-55, 2011

4 윤진숙, "눈개승마 추출물의 항산화 및 주름개선 효과" 한국식품저장유통학회 19 (19): 393-399, 2012

5 Kumar S, "α-glucosidase inhibitors from plants: A natural approach to treat diabetes" 5 : 19-29, 2011

6 Millauskas G, "Screening of radical scavenging activity of some medicinal and aromatic plant extracts" 85 : 231-237, 2004

7 Soobrattee MA, "Phenolics as potential antioxidant therapeutic agents:Mechanism and actions" 579 : 200-213, 2005

8 Barbosa Filho JM, "Natural products inhibitors of the enzyme acetylcholinesterase" 16 : 258-285, 2006

9 Hae Sook Baek, "Methanol Extract of Goat’s-beard (Aruncus dioicus) Reduces Renal Injury by Inhibiting Apoptosis in a Rat Model of Ischemia-Reperfusion" 한국식품영양과학회 17 (17): 101-108, 2012

10 Apostolidis E, "Inhibitory potential of herb, fruit, and fungal-enriched cheese against key enzymes linked to type 2 diabetes and hypertension" 8 : 46-54, 2007

11 De-Felice DFG, "Inflammation, defective insulin signaling, and mitochondrial dysfunction as common molecular denominators connecting type 2 diabetes to Alzheimer’s disease" 63 : 2262-2272, 2014

12 Yu T, "Increased production of reactive oxygen species in hyperglycemic conditions requires dynamic change of mitochondrial morphology" 103 : 2653-2658, 2005

13 Eruslanov E, "Identification of ROS using oxidized DCFDA and flow-cytometry" 594 : 57-72, 2009

14 Piwkowska A, "High glucose concentration affects the oxidant-antioxidant balance in cultured mouse podocytes" 112 : 1661-1672, 2011

15 Crane PK, "Glucose levels and risk of dementia" 369 : 540-548, 2013

16 Valko M, "Free radicals and antioxidants in normal physiological functions and human disease" 39 : 44-87, 2007

17 Ishige K, "Flavonoids protect neuronal cells from oxidative stress by three distinct mechanisms" 30 : 433-446, 2001

18 Butterfield DA, "Evidence that amyloid beta-peptide-induced lipid peroxidation and its sequelae in Alzheimer’s disease brain contribute to neuronal death" 23 : 655-664, 2002

19 Vincent AM, "Diabetic neuropathy: Cellular mechanisms as therapeutic targets" 7 : 573-583, 2011

20 Maritim AC, "Diabetes, oxidative stress, and antioxidants: A review" 17 : 24-38, 2003

21 Lee JM, "Contents of total flavonoid and biological activities of edible plants" 16 : 504-514, 2001

22 Lobner D, "Comparison of the LDH and MTT assays for quantifying cell death: Validity for neuronal apoptosis?" 96 : 147-152, 2000

23 Oboh G, "Comparative study on the inhibitory effect of caffeic and chlorogenic acids on key enzymes linked to Alzheimer’s disease and some pro-oxidant induced oxidative stress in rats’ brain-in vitro" 38 : 413-, 2013

24 Levin BE, "Brain glucose sensing and body energy homeostasis: role in obesity and diabetes" 276 : 1223-1231, 1999

25 Quoc Hung Vo, "Bioactive Constituents from the n-Butanolic Fraction of Aruncus dioicus var. kamtschaticus" 한국생약학회 20 (20): 274-280, 2014

26 Ak T, "Antioxidant and radical scavenging properties of curcumin" 174 : 27-37, 2008

27 Chang ST, "Antioxidant activity of extracts from Acacia confusa bark and heartwood" 49 : 3420-3424, 2001

28 Tesfaye S, "Advances in the epidemiology, pathogenesis and management of diabetic peripheral neuropathy" 28 : 8-14, 2012