돌산갓김치에서 분리된 glucosinolates의 항산화, 항고혈압, 항당뇨 및 항암활성에 관한 연구 (The) antioxidative, antihypertensive, antidiabetic and anticancer activities of separated glucosinolates from Dolsan leaf mustard (Brassica juncea) Kimchi원문보기
오선경
(Chonnam National University
Department of Biomedical and Electronic Engineering
국내박사)
갓 (leaf mustard, Brassica juncea)은 십자화과에 속하는 경엽 채소 류로 주로 김치로 식용되고 씨는 신미성 향신료로서 사용되고 있고, 식물성 2차 대사산물인 glucosinolates (GSLs)가 풍부하다. GSLs와 같은 분해산물들은 갓과 갓김치의 생리활성 물질로 추정되고 있으나 이와 관련된 연구는 미흡한 실정이다. 따라서, 본 연구에서는 Dolsan mustard seed (...
갓 (leaf mustard, Brassica juncea)은 십자화과에 속하는 경엽 채소 류로 주로 김치로 식용되고 씨는 신미성 향신료로서 사용되고 있고, 식물성 2차 대사산물인 glucosinolates (GSLs)가 풍부하다. GSLs와 같은 분해산물들은 갓과 갓김치의 생리활성 물질로 추정되고 있으나 이와 관련된 연구는 미흡한 실정이다. 따라서, 본 연구에서는 Dolsan mustard seed (DMS), Dolsan leaf mustard (DLM)과 Dolsan leafmustard kimchi (DLMK)에 함유되어있는 GSLs에 대한 동정 및 함량변화에 대한 분석과 GSLs의 성분분석 및 주요 isothiocyanate (ITC)의 항산화, 항고혈압, 항당뇨 및 암세포 생장억제효과에 대한 연구를 수행하였다. DMS 와DLM은 acetonitrile 추출법으로 추출하였다. DMS와 DLM의 조추출 물은 liquid chromatography-photodiode array detector 와 tandem mass spec trometry (LC-PDA/MS/MS)를 이용하여 8가지 GSLs성분을 동정하였으며, 4가지의 aliphatic GSLs (glucobrassicanapin, glucoraphanin, glucoiberverin, sinigrin)와 1가지의 acylated GSLs (glucoisatisin), 1가지의 indole GSLs (4-methoxyglucobrassicin)와 2가지의 aromatic GSLs (gluconasturtiin, glucotro paeolin)로 구성되어 있는 것으로 분석되었다. 이들의 총 GSLs함량은 DMS의 경우는 53.77 mg/g으로 나타났으며 생갓 부위에서는 5.17-6.35 mg/g으로 확인되었다. 0℃, 35일동안 발효하면서 7일간격으로 추출한 DLMK의 조추출물도 LC-PDA/MS/MS를 이용하여 총 4가지의 GSLs 성분 (glucobrassicin, sinigrin, glucotropaeolin, progotrin)을 확인하였다. 이들 성분 중 alphatic GSLs (sinigrin, progoitrin), indole GSLs (glucobrassicin), aromatic GSLs (glucotropaeolin)의 함량은 발효 14일째까지 증가하는 것으로 나타나는 반면, 발효 21일째 급격히 감소하는 경향을 나타냈다. 또한 DLMK에 존재하는 isothiocyanate (ITC)를 포함하는 휘발성 성분은 gas chromatography-mass spectrometry (GC-MS)로 측정하였다. DLMK의 발효과정 중 주요 휘발성 성분으로는 2-propenyl ITC (about 86.70% of peak areas), 3-butenyl ITC (4.46%), n-butyl ITC (0.70%), 2-phenylethyl ITC (2.70%)이 각각 분석되었다. GSLs 분해산물인 ITC 성분은 발효 14일까지 2-propenyl ITC (85.50%)과 3-butenyl ITC (4.50%)가 가장 높은 함량을 차지하는 것으로 확인되었으며, 발효 21일에 2-propenyl ITC과 3-butenyl ITC은 검출되지 않은 반면에 allylisothiocyanate (AITC)가 89.34%로 확인되었다. 이는 GSLs 함량이 발효기간에 따라 차이가 있는 것을 나타내는 결과이다. DMS, DLM 와DLMK 추출물은 preparative octadecysilane high performance liquid chromatography (ODS-HPLC)를 이용하여 peak별로 분리, 정제하여LC-PDA/MS/MS로 GSLs성분을 동정하였다. DMS에서 alphatic GSLs인sinigrin (2.55 mg/g)과glucobrassicanapin (0.45 mg/g)이 확인되었고, DLM의 뿌리에서 aromatic GSLs인 gluconasturtiin (0.48 mg/g), 줄기에서 alphatic GSLs인 glucobrassicanapin (0.49 mg/g), 잎에서는 acylated GSLs로 glucoiastisin (0.44 mg/g)과 aromatic GSLs인 glucotropaeolin (0.52 mg/g)이 확인되었다. DLMK에서 동정된 4가지 GSLs은 발효 기간 동안에 sinigrin (15.82-8.24 mg/g), glucobrassicin (0.71-0.40 mg/g), glucotropaeolin (0.50-0.39 mg/g), progotrin (1.20-1.85 mg/g)이 각각 확인되었다. DMS, DLM and DLMK 추출물에서 확인된 GSLs 각각의 성분을 대상으로 생리활성에 대한 연구를 진행하였다. 항산화 활성은 확인된 GSLs중에서 종자의 sinigrin이 높은 활성을 나타냈고, ACE 저해활성은 DLMK의 발효 14일째 sinigrin (71.60%)이 대조군인 0.01% captopril (78.30%)과 비슷한 저해활성이 나타냈고, α-glucosidase 저해활성 또한 DLMK의 발효 14일째 sinigrin (65.17%)이 대조군인 0.05% acarbose (50.83%)보다 높은 저해활성을 나타내었다. 또한 동정된 GSLs를 위암세포 SNU 601과 폐암세포 A 549에 대해서 농도별 (50-400 μg/mL)에 따른 암세포 생장억제효과를 측정하였다. 위암세포 SNU 601은 DMS의 sinigrin와 leaves의 glucotropaeolin (in DLM)에서 19.43-27.31%, 8.71-22.06%의 생장억제효과를 나타내었고, 폐암세포 A 549의 경우도 DMS의 sinigrin와 leaves의 glucotropaeolin (in DLM)에서 12.85-23.32%, 4.35-18.97%의 생장억제효과를 나타내었다. DLMK는 발효 14일째 암세포 생장억제효과를 나타냈고, SNU 601은 glucotropaeolin, A 549 는 sinigrin에서 10.24-19.74%, 11.93-29.34%의 생장억제효과를 나타내었다. 이러한 결과로 DMS, DLM, DLMK에서 확인된 GSLs와 주요 가수분해산물인 2-propenyl ITC와 3-butenyl ITC, AITC 의 함량변화는 항산화활성, ACE 저해활성, α-glucosidase저해활성 및 암세포 생장억제효과 등의 생리활성을 나타내는 것을 확인하였다.
갓 (leaf mustard, Brassica juncea)은 십자화과에 속하는 경엽 채소 류로 주로 김치로 식용되고 씨는 신미성 향신료로서 사용되고 있고, 식물성 2차 대사산물인 glucosinolates (GSLs)가 풍부하다. GSLs와 같은 분해산물들은 갓과 갓김치의 생리활성 물질로 추정되고 있으나 이와 관련된 연구는 미흡한 실정이다. 따라서, 본 연구에서는 Dolsan mustard seed (DMS), Dolsan leaf mustard (DLM)과 Dolsan leafmustard kimchi (DLMK)에 함유되어있는 GSLs에 대한 동정 및 함량변화에 대한 분석과 GSLs의 성분분석 및 주요 isothiocyanate (ITC)의 항산화, 항고혈압, 항당뇨 및 암세포 생장억제효과에 대한 연구를 수행하였다. DMS 와DLM은 acetonitrile 추출법으로 추출하였다. DMS와 DLM의 조추출 물은 liquid chromatography-photodiode array detector 와 tandem mass spec trometry (LC-PDA/MS/MS)를 이용하여 8가지 GSLs성분을 동정하였으며, 4가지의 aliphatic GSLs (glucobrassicanapin, glucoraphanin, glucoiberverin, sinigrin)와 1가지의 acylated GSLs (glucoisatisin), 1가지의 indole GSLs (4-methoxyglucobrassicin)와 2가지의 aromatic GSLs (gluconasturtiin, glucotro paeolin)로 구성되어 있는 것으로 분석되었다. 이들의 총 GSLs함량은 DMS의 경우는 53.77 mg/g으로 나타났으며 생갓 부위에서는 5.17-6.35 mg/g으로 확인되었다. 0℃, 35일동안 발효하면서 7일간격으로 추출한 DLMK의 조추출물도 LC-PDA/MS/MS를 이용하여 총 4가지의 GSLs 성분 (glucobrassicin, sinigrin, glucotropaeolin, progotrin)을 확인하였다. 이들 성분 중 alphatic GSLs (sinigrin, progoitrin), indole GSLs (glucobrassicin), aromatic GSLs (glucotropaeolin)의 함량은 발효 14일째까지 증가하는 것으로 나타나는 반면, 발효 21일째 급격히 감소하는 경향을 나타냈다. 또한 DLMK에 존재하는 isothiocyanate (ITC)를 포함하는 휘발성 성분은 gas chromatography-mass spectrometry (GC-MS)로 측정하였다. DLMK의 발효과정 중 주요 휘발성 성분으로는 2-propenyl ITC (about 86.70% of peak areas), 3-butenyl ITC (4.46%), n-butyl ITC (0.70%), 2-phenylethyl ITC (2.70%)이 각각 분석되었다. GSLs 분해산물인 ITC 성분은 발효 14일까지 2-propenyl ITC (85.50%)과 3-butenyl ITC (4.50%)가 가장 높은 함량을 차지하는 것으로 확인되었으며, 발효 21일에 2-propenyl ITC과 3-butenyl ITC은 검출되지 않은 반면에 allylisothiocyanate (AITC)가 89.34%로 확인되었다. 이는 GSLs 함량이 발효기간에 따라 차이가 있는 것을 나타내는 결과이다. DMS, DLM 와DLMK 추출물은 preparative octadecysilane high performance liquid chromatography (ODS-HPLC)를 이용하여 peak별로 분리, 정제하여LC-PDA/MS/MS로 GSLs성분을 동정하였다. DMS에서 alphatic GSLs인sinigrin (2.55 mg/g)과glucobrassicanapin (0.45 mg/g)이 확인되었고, DLM의 뿌리에서 aromatic GSLs인 gluconasturtiin (0.48 mg/g), 줄기에서 alphatic GSLs인 glucobrassicanapin (0.49 mg/g), 잎에서는 acylated GSLs로 glucoiastisin (0.44 mg/g)과 aromatic GSLs인 glucotropaeolin (0.52 mg/g)이 확인되었다. DLMK에서 동정된 4가지 GSLs은 발효 기간 동안에 sinigrin (15.82-8.24 mg/g), glucobrassicin (0.71-0.40 mg/g), glucotropaeolin (0.50-0.39 mg/g), progotrin (1.20-1.85 mg/g)이 각각 확인되었다. DMS, DLM and DLMK 추출물에서 확인된 GSLs 각각의 성분을 대상으로 생리활성에 대한 연구를 진행하였다. 항산화 활성은 확인된 GSLs중에서 종자의 sinigrin이 높은 활성을 나타냈고, ACE 저해활성은 DLMK의 발효 14일째 sinigrin (71.60%)이 대조군인 0.01% captopril (78.30%)과 비슷한 저해활성이 나타냈고, α-glucosidase 저해활성 또한 DLMK의 발효 14일째 sinigrin (65.17%)이 대조군인 0.05% acarbose (50.83%)보다 높은 저해활성을 나타내었다. 또한 동정된 GSLs를 위암세포 SNU 601과 폐암세포 A 549에 대해서 농도별 (50-400 μg/mL)에 따른 암세포 생장억제효과를 측정하였다. 위암세포 SNU 601은 DMS의 sinigrin와 leaves의 glucotropaeolin (in DLM)에서 19.43-27.31%, 8.71-22.06%의 생장억제효과를 나타내었고, 폐암세포 A 549의 경우도 DMS의 sinigrin와 leaves의 glucotropaeolin (in DLM)에서 12.85-23.32%, 4.35-18.97%의 생장억제효과를 나타내었다. DLMK는 발효 14일째 암세포 생장억제효과를 나타냈고, SNU 601은 glucotropaeolin, A 549 는 sinigrin에서 10.24-19.74%, 11.93-29.34%의 생장억제효과를 나타내었다. 이러한 결과로 DMS, DLM, DLMK에서 확인된 GSLs와 주요 가수분해산물인 2-propenyl ITC와 3-butenyl ITC, AITC 의 함량변화는 항산화활성, ACE 저해활성, α-glucosidase저해활성 및 암세포 생장억제효과 등의 생리활성을 나타내는 것을 확인하였다.
Leaf mustard (Brassica juncea), a leaf and stem vegetable, belongs to the Cruciferae family. The stems and leaves from this plant are widely used as the main ingredient of leaf mustard kimchi and the seeds from this plant are used as acrid spices. Glucosinolates (GSLs) are a family of secondary meta...
Leaf mustard (Brassica juncea), a leaf and stem vegetable, belongs to the Cruciferae family. The stems and leaves from this plant are widely used as the main ingredient of leaf mustard kimchi and the seeds from this plant are used as acrid spices. Glucosinolates (GSLs) are a family of secondary metabolites, abundant in the seeds and green tissues of this plant. It has been suggested that GSLs and its hydrolysated products comprise a distinctive group of bioactive compounds. However, not many studies have investigated the content of GSL and its major hydrolysis product, isothiocyanates (ITCs) in Dolsan mustard seed (DMS), Dolsan leaf mustard (DLM), and Dolsan leaf mustard kimchi (DLMK) and also their antioxidant, antihypertensive, antidiabetic, and anticancer properties. DMS, different parts of DLM, and DLMK were subjected to acetonitrile extraction. Eight major GSLs were identified and quantified from the crude extract of DMS and DLM by using liquid chromatography-photodiode array detector and tandem mass spectrometry (LC-PDA/MS/MS). The GSLs consisted of four aliphatic GSLs (glucobrassicanapin, glucoraphanin, glucoiberverin, and sinigrin); one acylated GSL (glucoisatisin); one indole GSL (4-hydroxyglucobrassicin); and aromatic GSLs (gluconasturtiin and glucotropaeolin). The total GSL content in DMS was measured to be 53.77 mg/g, and the GSL content in different parts of DLM ranged between 5.17 and 6.35 mg/g. At 0°C, four components of GSLs (glucobrassicin, sinigrin, glucotropaeolin, and progotrin) were detected in the crude extracts of DLMK, by using LC-PDA/MS/MS. Extractions were performed every 7 days, during the fermentation period of 35 days. The content of sinigrin and progotrin (aliphatic GSLs), glucobrassicin (indole GSL), glucotropaeolin (aromatic GSL) increased until fermentation day 14. However, the content of these GSLs rapidly decreased during 21 days of fermentation. In addition, volatile components, including ITC, present in DLMK was measured using gas chromatography-mass spectrometry (GC-MS). The main Jovolatile components of DLMK were found to be 2-propenyl ITC (about 86.70% of peak areas), 3-butenyl ITC (4.46%), n-butyl ITC (0.70%), and 2-phenylethyl ITC (2.70%). The highest content of the major hydrolysis products of GSLs, 2-propenyl ITC (85.50%) and 3-butenyl ITC (4.50%) were detected at fermentation day 14. However, during the fermentation period of 21 days, 2-propenyl ITC and 3-butenyl ITC were not detected but high amounts of allylisothiocyanate (AITC) (89.34%) were detected. Therefore, these results indicated that the content of GSLs varied depending on the duration of fermentation. The GSL components from the crude extracts of DMS, different parts of DLM, and DLMK were separated using preparative octadecysilane high-performance liquid chromatography (ODS-HPLC). The identified components of GSLs were purified using LC-PDA/MS/MS. The acylated GSLs were found to contain sinigrin (2.55 mg/g) and glucobrassicanapin (0.45 mg/g) (aliphatic GSLs) in DMS, gluconasturtiin (0.48 mg/g) (aromatic GSL) in the root of DLM, glucobrassicanapin (0.49 mg/g) (aliphatic GSL) in the stem, glucoisatisin (0.44 mg/g) and glucotropaeolin (0.52 mg/g) (aromatic GSL) in the leaves. Four GSLs that were detected in DLMK during the fermentation periods were sinigrin (15.82-8.24 mg/g), glucobrassicin (0.71-0.40 mg/g), glucotropaeolin (0.50-0.39 mg/g), and progotrin (1.20-1.85 mg/g). This study was conducted to delineate the physiological activity of each of the identified components of GSLs isolated from DMS, different parts of DLM, and DLMK extract. The analysis of antioxidant activity, confirmed that sinigrin, a component of GSL (DMS) showed high antioxidant activity. In DLMK, during the fermentation period of 14 days, sinigrin showed ACE (angiotensin converting enzyme) inhibitory activity (71.60%) similar to that shown by 0.01% captopril, the control group (78.30%). Similarly, during the fermentation period of 14 days, sinigrin exhibited high α-glucosidase inhibitory activity (65.17%) compared to that observed for 0.05% acarbose, the control group (50.83%). The cancer cell growth inhibitory effect of GSLs was analyzed at various concentrations (50-400 μg/mL) in SNU 601 (gastric cancer cell line) and A 549 (lung cancer cell line). The percentage of growth inhibition caused by sinigrin (DMS) and glucotropaeolin (DLM leaves) ranged between 19.43 and 27.31% and between 8.71 and 22.06% in SNU 601 and A 549 cells, respectively. Similarly, in A 549 cells, the percentage of growth inhibition caused by sinigrin and glucotropaeolin (DLM leaves) ranged between 12.85 and 23.32% and between 4.35 and 18.97%, respectively. In DLMK, during the fermentati`on period of 14 days, glucotropaeolin and sinigrin showed growth inhibitory effects against SNU 601 and A 549 cells, respectively. The percentage of inhibition caused by glucotropaeolin and sinigrin ranged between 10.24 and 19.74% and between 11.93 and 29.34%, respectively. In the present study, GSLs and their major hydrolysated products, 2-propenyl ITC, and 3-butenyl ITC, in DMS, different parts of DLM, and DLMK, were investigated. The content of allyl iosthiocyanate (AITC) changed with varying periods of fermentation. In this study, the various components of GSLs were investigated for their antioxidant, antihypertensive, antidiabetic, and anticancer properties.
Leaf mustard (Brassica juncea), a leaf and stem vegetable, belongs to the Cruciferae family. The stems and leaves from this plant are widely used as the main ingredient of leaf mustard kimchi and the seeds from this plant are used as acrid spices. Glucosinolates (GSLs) are a family of secondary metabolites, abundant in the seeds and green tissues of this plant. It has been suggested that GSLs and its hydrolysated products comprise a distinctive group of bioactive compounds. However, not many studies have investigated the content of GSL and its major hydrolysis product, isothiocyanates (ITCs) in Dolsan mustard seed (DMS), Dolsan leaf mustard (DLM), and Dolsan leaf mustard kimchi (DLMK) and also their antioxidant, antihypertensive, antidiabetic, and anticancer properties. DMS, different parts of DLM, and DLMK were subjected to acetonitrile extraction. Eight major GSLs were identified and quantified from the crude extract of DMS and DLM by using liquid chromatography-photodiode array detector and tandem mass spectrometry (LC-PDA/MS/MS). The GSLs consisted of four aliphatic GSLs (glucobrassicanapin, glucoraphanin, glucoiberverin, and sinigrin); one acylated GSL (glucoisatisin); one indole GSL (4-hydroxyglucobrassicin); and aromatic GSLs (gluconasturtiin and glucotropaeolin). The total GSL content in DMS was measured to be 53.77 mg/g, and the GSL content in different parts of DLM ranged between 5.17 and 6.35 mg/g. At 0°C, four components of GSLs (glucobrassicin, sinigrin, glucotropaeolin, and progotrin) were detected in the crude extracts of DLMK, by using LC-PDA/MS/MS. Extractions were performed every 7 days, during the fermentation period of 35 days. The content of sinigrin and progotrin (aliphatic GSLs), glucobrassicin (indole GSL), glucotropaeolin (aromatic GSL) increased until fermentation day 14. However, the content of these GSLs rapidly decreased during 21 days of fermentation. In addition, volatile components, including ITC, present in DLMK was measured using gas chromatography-mass spectrometry (GC-MS). The main Jovolatile components of DLMK were found to be 2-propenyl ITC (about 86.70% of peak areas), 3-butenyl ITC (4.46%), n-butyl ITC (0.70%), and 2-phenylethyl ITC (2.70%). The highest content of the major hydrolysis products of GSLs, 2-propenyl ITC (85.50%) and 3-butenyl ITC (4.50%) were detected at fermentation day 14. However, during the fermentation period of 21 days, 2-propenyl ITC and 3-butenyl ITC were not detected but high amounts of allylisothiocyanate (AITC) (89.34%) were detected. Therefore, these results indicated that the content of GSLs varied depending on the duration of fermentation. The GSL components from the crude extracts of DMS, different parts of DLM, and DLMK were separated using preparative octadecysilane high-performance liquid chromatography (ODS-HPLC). The identified components of GSLs were purified using LC-PDA/MS/MS. The acylated GSLs were found to contain sinigrin (2.55 mg/g) and glucobrassicanapin (0.45 mg/g) (aliphatic GSLs) in DMS, gluconasturtiin (0.48 mg/g) (aromatic GSL) in the root of DLM, glucobrassicanapin (0.49 mg/g) (aliphatic GSL) in the stem, glucoisatisin (0.44 mg/g) and glucotropaeolin (0.52 mg/g) (aromatic GSL) in the leaves. Four GSLs that were detected in DLMK during the fermentation periods were sinigrin (15.82-8.24 mg/g), glucobrassicin (0.71-0.40 mg/g), glucotropaeolin (0.50-0.39 mg/g), and progotrin (1.20-1.85 mg/g). This study was conducted to delineate the physiological activity of each of the identified components of GSLs isolated from DMS, different parts of DLM, and DLMK extract. The analysis of antioxidant activity, confirmed that sinigrin, a component of GSL (DMS) showed high antioxidant activity. In DLMK, during the fermentation period of 14 days, sinigrin showed ACE (angiotensin converting enzyme) inhibitory activity (71.60%) similar to that shown by 0.01% captopril, the control group (78.30%). Similarly, during the fermentation period of 14 days, sinigrin exhibited high α-glucosidase inhibitory activity (65.17%) compared to that observed for 0.05% acarbose, the control group (50.83%). The cancer cell growth inhibitory effect of GSLs was analyzed at various concentrations (50-400 μg/mL) in SNU 601 (gastric cancer cell line) and A 549 (lung cancer cell line). The percentage of growth inhibition caused by sinigrin (DMS) and glucotropaeolin (DLM leaves) ranged between 19.43 and 27.31% and between 8.71 and 22.06% in SNU 601 and A 549 cells, respectively. Similarly, in A 549 cells, the percentage of growth inhibition caused by sinigrin and glucotropaeolin (DLM leaves) ranged between 12.85 and 23.32% and between 4.35 and 18.97%, respectively. In DLMK, during the fermentati`on period of 14 days, glucotropaeolin and sinigrin showed growth inhibitory effects against SNU 601 and A 549 cells, respectively. The percentage of inhibition caused by glucotropaeolin and sinigrin ranged between 10.24 and 19.74% and between 11.93 and 29.34%, respectively. In the present study, GSLs and their major hydrolysated products, 2-propenyl ITC, and 3-butenyl ITC, in DMS, different parts of DLM, and DLMK, were investigated. The content of allyl iosthiocyanate (AITC) changed with varying periods of fermentation. In this study, the various components of GSLs were investigated for their antioxidant, antihypertensive, antidiabetic, and anticancer properties.
주제어
#Dolsan leaf mustard kimchi glucosinolates
학위논문 정보
저자
오선경
학위수여기관
Chonnam National University
학위구분
국내박사
학과
Department of Biomedical and Electronic Engineering
AI-Helper
※ AI-Helper는 부적절한 답변을 할 수 있습니다.