제주도 탈염 용암 해수를 이용하여 수경재배한 새싹인삼 추출물의 In Vitro 신경독성에 대한 신경보호 효과

국문 초록 (Abstract)

본 연구에서는 유용한 미네랄 성분을 다량 함유하고 있어 산업적 가치가 높은 제주 용암 해수에서 탈염 과정을 거쳐 만들어진 탈염 용암 해수를 이용하여 수경재배한 새싹인삼추출물 건조시료를 Q-Orbitrap으로 성분을 규명하고, HPLC로 기준 및 시험방법을 설정하여 표준화했으며 ICP-AES를 이용하여 무기물 함량을 분석하였다. 시험결과에서 주요 ginsenoside Rb1, Rg1 및 Na, Mg, Ca, Sr, V와 같은 미네랄 함량이 증가함을 확인했으며 이에 대한 효능을 입증하기 위해서 제주도 탈염 용암 해수를 이용해 수경재배한 새싹인삼 추출물에 대한 신경세포사멸과 신경세포 염증 반응 억제, 신경전달물질 및 시냅스 가소성 회복을 통한 기억력 개선에 대해 in vitro 효능검증을 수행하였다. 제주도 탈염 용암 해수 수경재배 새싹인삼 추출물에 대한 동결 건조를 진행 후 ginsenoside 함량 분석을 진행한 결과에서 ginsenoside Rb1(11.5 mg/g), Rg1(6.85 mg/g), Re(19.2 mg/g)의 합이 37.55 mg/g으로 측정되었고, 이는 식약처 건강기능식품 인삼 기능성분의 함량기준인 Rb1, Rg1의 합이 0.8~34 mg/g인 것과 비교했을 때, 탈염 용암 해수로 수경재배한 제주새싹인삼 동결 건조물은 개별 인정형 원료로서의 충분한 요건을 갖춘 것으로 사료된다. 또한 제주도 탈염 용암 해수 수경재배 새싹인삼 추출물 건조시료에 대한 세포독성 및 효능평가 결과에서 제주도 탈염 용암 해수 수경재배 새싹인삼추출물 건조시료는 RA-differentiated SH-SY5Y neuronlike cell에 48시간 동안 처리 시 1,250 μg/mL에서도 세포독성을 보이지 않았으며, 더욱이 Aβ1-42 oligomer(10 μM)를 72시간 동안 처리하여 세포독성을 유도한 세포실험 모델에서 비처리 대조군과 비교하여 제주도 탈염 용암 해수 수경재배 새싹인삼 추출물 건조시료를 처리한 실험군에서 세포사멸 관련 바이오마커들인 cleaved/total caspase-3 ratio, cleaved/total PARP, Bax/Bcl-2 ratio를 감소시켜 Aβ1-42 oligomer에 의한 신경세포사멸을 억제하는 효과를 보였다. 그리고 RA-differentiated SH-SY5Y neuron-like cell에 H₂O₂(300 μM)를 처리하여 신경세포 염증 반응을 유도한 세포실험 모델에서 비처리 대조군과 비교하여 제주도 탈염용암 해수 수경재배 새싹인삼 추출물 건조시료를 처리한 실험군에서 염증 반응과 관련된 NF-κB p65 단백질의 nuclear translocation 억제, COX-2 단백질 발현 감소, MAPK (ERK1/2, SAPK/JNK, p38 MAPK) 인산화를 감소시켜 H₂O₂에 의한 신경세포 염증 반응을 억제하는 효과를 보였다. 한편 RA-differentiated SH-SY5Y neuron-like cell에 Aβ1-42 oligomer(1 μM)를 처리하여 신경전달물질 및 시냅스 가소성 감소를 유도한 세포실험 모델에서 비처리 대조군과 비교하여 제주도 탈염 용암 해수 수경재배 새싹인삼 추출물 건조시료를 처리한 실험군에서 시냅스 가소성과 관련된 CREB 전사인자의 인산화를 통한 활성화로 신경전달물질인 BDNF 발현이 증가하는 효과를 보였으며, AChE 활성을 저해함으로써 Aβ1-42 oligomer에 의한 신경전달물질 및 시냅스 가소성 감소를 억제하는 효과를 보였다. 그러므로 제주도 탈염 용암 해수를 이용한 새싹인삼은 질환 특이적인 성분의 증가를 높여줌으로써 산업적 이용가치가 높을 것으로 판단되며, 이를 활용한 다른 약용작물 및 특용작물의 생산을 통한 건강기능식품 원료개발에도 충분히 활용 가능할 것으로 사료된다.

다국어 초록 (Multilingual Abstract)

This study investigated the neuroprotective effects of extract of Panax ginseng sprouts cultivated by hydroponics using desalinated magma seawater from Jeju island (Korea) on neurotoxicity in vitro. The chemical compositions of the Panax ginseng sprouts extracts were analyzed using LC-Q-orbitrap, and ginsenosides (Re, Rg1, Rg2, Rb1, and Rd) were detected. Analysis method based on HPLC was developed for determination of ginsenosides Rb1, Rg1, and Re. For the in vitro analysis, the extract inhibited Aβ1-42 oligomer-induced neurotoxicity through inhibition of cleaved caspase-3, cleaved PPAR, and Bax expression, as well as activation of Bcl-2 expression. The extract inhibited H₂O₂-induced neuroinflammatory responses through inhibition of nuclear translocation of NF-κB p65 and COX-2 expression, in addition to phosphorylation of MAPK (ERK1/2, SAPK/JNK, and p-38 MAPK). The extract stimulated BDNF expression through CREB activation by phosphorylation at Ser<SUP>133</SUP> and inhibited up-regulation of AChE activity by Aβ1-42 oligomer-induced neurotoxicity. Taken together, these results provide in vitro evidence that the experimental extract exhibits neuroprotective properties against Aβ1-42-induced neurotoxicity and oxidative stress, which suggests that the extract has potential to be developed into a new functional food for neuroprotection and learning/memory improvement in the global food market. Furthermore, it is expected that desalinated magma seawater from Jeju island (Korea) may be applied for the cultivation of other functional plants in order to enhance functional components.

목차 (Table of Contents)

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

참고문헌 (Reference)

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