[논문]스마트팜 재배 병풀의 triterpenes 정량 및 각질형성세포 활성화 효과

박진홍; 조성민; 이다희; 박영민; 장환봉; 강태진; 이기만

doi:10.11002/kjfp.2023.30.3.483

스마트팜 재배 병풀의 triterpenes 정량 및 각질형성세포 활성화 효과
Quantification of triterpenes in Centella asiatica cultivated in a smart farm, and their effect on keratinocyte activation 원문보기

한국식품저장유통학회지 = Korean journal of food preservation, v.30 no.3, 2023년, pp.483 - 491

박진홍 (록야 주식회사, 그린바이오연구소) , 조성민 (록야 주식회사, 그린바이오연구소) , 이다희 (록야 주식회사, 그린바이오연구소) , 박영민 (록야 주식회사, 그린바이오연구소) , 장환봉 (록야 주식회사, 그린바이오연구소) , 강태진 (삼육대학교 약학대학) , 이기만 (록야 주식회사, 그린바이오연구소)

초록
AI-Helper

본 연구에서는 제주도에서 자생한 병풀을 수집해 스마트팜과 노지에서 재배하고 이를 이용하여 주요성분 및 각질형성세포 활성화에 미치는 영향을 확인 및 비교하였다. 스마트팜 재배 병풀과 노지 재배 병풀의 유전자 확인을 통한 종 분석을 위해, 핵 속의 ITS DNA와 엽록체의 psbA-H DNA를 증폭하여 염기서열을 분석한 후 NCBI 유전자 은행에서 보고된 식물들의 DNA와 비교하였다. 스마트팜 재배 병풀과 노지 재배 병풀의 ITS DNA 염기서열은 유전자 은행의 MH768338.1번 Centella asiatica와 일치하고 엽록체 psbA-H DNA 또한 유전자 은행의 JQ425422.1번 C. asiatica와 일치하였다. 스마트팜 재배 병풀추출물(SEE)과 노지 재배 병풀추출물(FEE)의 triterpene은 HPLC에 의해 분석되었으며, SEE의 madecassoside, asiaticoside, madecassic acid, asiatic acid 함량은 각각 59.31±0.94 mg/g, 46.38±2.26 mg/g, 6.21±1.47 mg/g, 7.04±1.93 mg/g으로 분석되었다. 반면, FEE는 각각 24.38±1.31 mg/g, 21.28±1.44 mg/g, 3.11±1.05 mg/g, 5.40±1.26 mg/g으로 측정되어 SEE가 FEE보다 더 높은 triterpene을 갖는 것이 확인되었다. 사람 각질형성세포에 대한 SEE와 FEE의 독성은 실험된 농도 내에서 관찰되지 않았으며, 스크래치가 유발된 세포 내 회복은 SEE가 FEE보다 더 높은 회복능을 보였다. 따라서, 본 실험 결과 triterpene 함량이 더 높은 스마트팜 재배 병풀이 건강기능식품 소재로서 더 효과적이라고 판단된다.

Abstract ▼ AI-Helper

This study aimed to compare the bioactive compounds in Centella asiatica (C. asiatica) cultivated in a smart farm and a field and their effects on human keratinocyte cells. C. asiatica was collected in Jeju-do, Korea, and cultured in a smart farm and a field. The main bioactive compounds in the two differentially cultured C. asiatica were identified, and their activation in keratinocytes were assessed. Amplification and sequencing of the internal transcribed spacer (ITS) DNA in the nucleus and psbA-H DNA in the chloroplast were performed for species analysis. A comparison of DNA of plants reported in the NCBI GenBank was performed. The ITS DNA and psbA-H DNA sequences of C. asiatica cultivated in a smart farm and a field were consistent with No. MH768338.1 and No. JQ425422.1, respectively. Analysis of the triterpenes was performed using high performance liquid chromatography (HPLC) and as a result, C. asiatica cultured in a smart farm had more triterpenes than those cultured in a field. The effects of C. asiatica grown in a smart farm on cell proliferation and scratch recovery in HaCaT cells were greater than those grown in a field. These results suggest that C. asiatica cultivated in a smart farm can be effectively utilized as a health functional food.

주제어

표/그림 (7)

표 Table 1. Calibration curve and linearity of standards of triterpenes
그림 Fig. 1. PCR result of Centella asiatica. M, marker; N, negative control. (A) PCR result of Centella asiatica cultivated in a smart farm, (B) PCR result of Centella asiatica cultivated in a field.
그림 Fig. 2. HPLC chromatogram of triterpenes in Centella asiatica. Standard mixture and samples were applied to the HPLC system at dose of 250 μg/mL, and 10 mg/mL, respectively. (A) HPLC chromatogram of standard mixture, (B) HPLC chromatogram of triterpenes in SEE, (C) HPLC chromatogram of triterpenes in FEE. SEE, 70% ethanol extract of Centella asiatica cultivated in a smart farm; FEE, 70% ethanol extract of Centella asiatica cultivated in a field.
표 Table 2. Genetic similarity of Centella asiatica with ITS and psbA-H region in GenBank
그림 Fig. 3. Cytotoxicity of SEE and FEE in HaCaT cell. Cells were treated with different concentration of SEE and FEE, respectively. After 24 hours, viability was measured with WST assay. Results were shown as percentage of untreated group. The results were the means±SD (n=3). p-values were compared with the untreated group at *p<0.05, **p<0.01 using the GraphPad Prism statistical package. (A) SEE, (B) FEE.
표 Table 3. Concentrations of major triterpenes from Centella asiatica
그림 Fig. 4. Effects of SEE and FEE on scratch recovery in HaCaT cell. HaCaT cells were seeded at 2×105 cells/well in 24-well plates and cultured at 37℃ for 24 h. After the monolayer of HaCaT cells had been established, scratches were induced using a sterile 200 μL pipet tip. Then, 200 μg/mL of SEE and FEE and 20 ng/mL of TNF-α were treated, respectively. Images were captured by day 3. Wound closure ratio was calculated by the following equation. Wound closure ratio=[(Surface area of day 0 of treated group)-(Surface area of each day of treated group)/(Surface area of day 0 of untreated group)-(Surface area of each day of untreated group)]. Results were the means±SD (n=3). p-values were compared with the untreated group at *p<0.05, **p<0.01 using the GraphPad Prism statistical package. (A) Image of HaCaT cell migration, (B) wound closure ratio in HaCaT cell.

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