Improvement of Biological activities of Acer mono and Acer okamotoanum Saps by Nano-encapsulation Process

나노입자화 공정을 이용한 고로쇠 및 우산고로쇠 수액의 유용생리활성 증진

  • Jeong, Myoung-Hoon (College of Bioscience & Biotechnology, Kangwon National University) ;
  • Ha, Ji-Hye (College of Bioscience & Biotechnology, Kangwon National University) ;
  • Oh, Sung-Ho (College of Bioscience & Biotechnology, Kangwon National University) ;
  • Kim, Seung-Seop (College of Bioscience & Biotechnology, Kangwon National University) ;
  • Jin, Ling (College of Bioscience & Biotechnology, Kangwon National University) ;
  • Lee, Hak-Ju (Korea Forest Research Institute) ;
  • Kang, Ha-Young (Korea Forest Research Institute) ;
  • Prak, Uk-Yeon (Department of Food Processing & Bakery, Gangneung Provincial University) ;
  • Lee, Hyeon-Yong (College of Bioscience & Biotechnology, Kangwon National University,Research Institute of Bioscience and Biotechmology, Kangwon National University)
  • 정명훈 (강원대학교 BT특성화학부(대학)) ;
  • 하지혜 (강원대학교 BT특성화학부(대학)) ;
  • 오성호 (강원대학교 BT특성화학부(대학)) ;
  • 김승섭 (강원대학교 BT특성화학부(대학)) ;
  • 김영 (강원대학교 BT특성화학부(대학)) ;
  • 이학주 (국립산림과학원) ;
  • 강하영 (국립산림과학원) ;
  • 박욱연 (강원도립대학 식품가공제과제빵과) ;
  • 이현용 (강원대학교 BT특성화학부(대학),강원대학교 생명공학연구소)
  • Received : 2009.04.27
  • Accepted : 2009.06.19
  • Published : 2009.09.30

Abstract

We investigated the improvement of immuno-modulatory activities of sap of Acer mono and A. okamotoanum encapsulated with edible polymers. Anticancer activities and immune activities such as human B and T cell growth, secretion of cytokines and natural killer cell growth were observed. Both human immune B and T cells were increased up to 30~50% by the addition of nano particle sap of Acer mono and A. okamotoanum. The secretion of Interleukin-6 (IL-6) and Tumor necrosis factor-a (TNF-a) from human immune B and T cells were also significantly increased compare to the control. Natural Killer (NK) cell growth was enhanced to $19.4{\times}10^5$ cells/mL in adding nano encapsulated sap of A.okamotoanum. The cytotoxicity of the sample on normal human lung cell (HEL299) was below 19.8% in adding 1.0 mg/mL of the nano particle sap of A. okamotoanum. Generally, the growth of all three human lung adenocarcinoma, human stomach adenocarcinoma and human liver adenocarcinama was inhibited up to 85% in adding 1.0 mg/mL of the encapsulated sap. Interestingly enough, the encapsulated sap was completely penetrated into human cancer cells within 30 min after addition. It showed that the encapsulation of the sap definitely increased its biological activities, which can expand its use to wide range of food industries.

본 연구는 고로쇠 및 우산고로쇠 수액의 나노입자화를 통한 유용생리활성 증진에 관한 것이다. 먼저 나노입자 수액의 형태 및 크기 확인을 위하여 EF-TEM을 이용한 결과 유상이 주머니처럼 수액을 포집한 리포좀 형태를 띄고 있으며 약 200 nm 크기의 구형으로 형성된 것을 확인할 수 있었으며, 나노입자의 크기와 균일성 및 크기별 분포 측정을 위해 DLS를 이용한 결과 수십 nm에서 수천 nm의 크기를 보였고 그중 약 70%가 100~300 nm의 크기로 형성된 것을 확인하였다. 다음으로 나노입자의 세포독성 실험을 실시하였다. 일반 고로쇠 및 우산고로쇠 수액과 비교하여 큰 차이를 보이지 않았다. 이를 통해 나노입자화를 통한 세포독성의 위험은 없다고 판단하여, 인간 면역세포인 B 세포와 T 세포의 생육증진 실험을 하였다. 5일째 되는 날 가장 많은 세포의 수를 확인할 수 있었고 고로쇠 수액 보다는 우산고로쇠 수액의 면역세포 생육증진효과가 좋았다. 또한 일반 수액보다는 나노입자수액의 생육증진 효과가 크게 나타났다. Cytokine 분비량 및 NK 세포 활성의 경우에서도 위의 실험과 비슷한 결과를 얻을 수 있었다. 면역활성을 확인한 결과를 바탕으로 정상세포 독성 및 항암실험을 하였다. 인간 정상 폐세포인 HEL299에 대한 세포독성 실험 결과 수액 및 나노입자 수액 두 시료 전부 19%이하의 세포독성을 보임으로써 레시틴으로 포집한 수액의 정상세포에서의 안전성을 확인할 수 있었다. AGS, A549, Hep3B에 대한 암세포 생육억제실험결과 3가지 암세포에서 모두 수액에 비해 나노입자수액의 활성이 약 20% 증가 된 것을 확인할 수 있었다. 본 연구에서 수행한 일반 수액 및 나노입자 수액의 연구를 통하여 나노입자 수액의 기능성 소재로서의 활용성을 확인할 수 있었으며, 이를 바탕으로 더욱 다양한 생리활성 연구 및 유용생리활성 물질의 추출에 관한 연구가 이루어져야 할 것으로 생각된다.

Keywords

Acknowledgement

Supported by : 국립산림과학원

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