상황버섯 균사체로 고체 발효한 수삼의 지질 저하 효과 = Lipid-lowering effect of solid fermented Panax ginseng by Phellinus linteus mycelium

다국어 초록 (Multilingual Abstract)

In this study, fresh Panax ginseng (80.09% moisture content) was solid fermented with mycelium to increase lipid lowering ability. Fresh ginseng (FG) was cultured by inoculating 20% (w/v) liquid spawn of Phellinus linteus mycelium in the incubator (27±1℃, 80±5% humidity) for 30 days. Hot water extracts of Panax ginseng were prepared for in vivo and in vitro studies.
After fermentation, total phenolic and reducing sugar content of FG were increased by 62.2 and 24.7%, respectively (p<0.05). Radical scavenging activities of both FG and solid fermented fresh ginseng (SFFG) were negligible. Amounts of ginsenosides in FG by fermentation were changed significantly. Ginsenoside Rg1, Rb1 and Re contents were increased by 35.0, 94.5 and 42.1%, respectively, while Rh1, Rc, Rg3, and Rh2 contents were decreased by 96.1, 20.2, 97.5 and 100%, respectively, after fermentation. ICR mice fed high fat diet (40% lard, w/w, HFD) for 4 weeks were divided in to three groups (n=10 for each group) based on body weight and plasma triglyceride concentration. Each group was fed HFD continuously with oral administration of either hot water extracts of FG or SFFG (100 mg/Kg body weight/day). Control group fed distilled water as a vehicle. Compared with CON group, plasma TG concentrations of FG and SFFG were decreased by 19.9 (p<0.05) and 29.8% (p<0.01), respectively. TG concentration was further lowered by 12.6% (p<0.05) after fermentation. Total cholesterol concentrations of FG and SFFG were decreased by 11.4 (p<0.05), 30.8% (p<0.01), respectively. By fermentation, plasma TC concentration of FG was further decreased by 22.0% (p<0.01). Also LDL-C for these groups were reduced by 23.8 (p<0.01), 54.8% (p<0.001), respectively, compared with those of CON group. LDL concentration for SFFG were lower by 40.7% (p<0.001) than that of FC. AST levels of FG and SFFG groups elevated by high fat diet were returned to the normal range. Hepatic TG concentration for FG and SFFG groups were decreased by 14.5 and 25.3% (p<0.05), respectively. Hepatic TC concentrations for FG and SFFG groups were lowered by 23.5 (p<0.01) and 25.8% (p<0.001) than those of CON group. Protein concentration of FAS and SREBP-1 for FG and SFFG group were decreased by 25.0 (p=0.002) and 30.1% (p=0.001), and 15.8 and 16.5%, respectively, compared with those of CON group. Protein concentrations of HMGCR and SREBP-2 were reduced by 29.1 (p<0.001), 45.5% (p<0.001), and 26.7 (p=0.03), 35.3% (p=0.03), compared with those of CON group. After fermentation, protein concentrations of HMGCR and SREBP-2 of FG group were further decreased down by 23.1% (p=0.02) and 11.7% (p=0.05), respectively.
In conclusion, solid fermentation of fresh ginseng with Phellinus linteus mycelium was succeeded in terms of elevating lipid lowering ability via changing the ginsenosides contents during the fermentation. Certain ginsenosides such as Rg1, Rb1 and Re concentrations were increased while Rh1, Rc, Rg3, and Rh2 were decreased. Suppression of plasma and hepatic cholesterol concentration by solid fermented fresh ginseng were noticeable. One of possible explanation of these effects might be due to decreased HMG-Co A reductase activity via suppressing SREBP-2 expression by SFFG.

목차 (Table of Contents)

• Ⅰ. 서 론 1
• Ⅱ. 재료 및 방법
• 1. 상황버섯 균사체를 이용한 수삼의 고체 발효 5
• 1) 버섯 균사체 배양 5
• 2) 액체 종균 제조 5
• 3) 고체 발효 6
• 4) 고체 발효 수삼의 열수 추출물 제조 7
• 2. 추출물의 이화학적 특성 분석 및 유리기 소거능 측정 7
• 1) 시약 및 기기 7
• 2) 총 페놀 함량 8
• 3) 환원당 함량 8
• 4) 유리기 소거능 8
• (1) DPPH radical 8
• (2) Superoxide anion radical 9
• (3) Hydroxyl radical 9
• (4) Nitrite radical 10
• 3. Ginsenosides 함량 분석 10
• 4. 고체 발효 수삼의 혈중 지질 저하 효과 관련 동물실험 11
• 1) 식이제조 11
• 2) 실험동물 및 사육 11
• 3) 해부 및 시료채취 13
• 4) 혈액 및 간의 생화학적 특성 분석 13
• (1) 혈중 지질 농도 측정 13
• ① 중성지방 농도 13
• ② 총 콜레스테롤 농도 14
• ③ HDL cholesterol 농도 14
• ④ LDL cholesterol 농도 14
• (2) AST, ALT 농도 측정 15
• (3) 간의 지질 농도 측정 15
• 5) Western blot analysis로 지질 대사 관련 효소 및 전사인자 발현 정도 측정 16
• (1) 시료 제조 16
• (2) 지질 대사 관련 효소 발현 정도 측정 17
• ① Fatty acid synthase 단백질 발현 17
• ② HMG-Co A reductase 단백질 발현 18
• (3) 지질 대사 관련 핵 전사인자 발현 정도 측정 18
• ① SREBP-1의 단백질 농도 18
• ② SREBP-2의 단백질 농도 19
• 5. 통계처리 19
• Ⅲ. 결과 및 고찰
• 1. 상황버섯 액체 종균 배양 21
• 2. 수삼의 고체 발효 양상 21
• 3. 고체 발효 수삼의 이화학적 특성 변화 24
• 1) 추출 수율 및 총 페놀, 환원당 함량 24
• 2) 고체 발효에 의한 수삼의 유리기 소거 효과 26
• (1) DPPH radical 소거 효과 26
• (2) Superoxide anion radical 소거 효과 28
• (3) Hydroxyl radical 소거 효과 30
• (4) Nitrite radical 소거 효과 32
• 4. 고체 발효에 의한 수삼 활성 성분의 변화 35
• 5. 고체 발효 수삼이 고지방식이를 섭취한 ICR mice의 혈중 지질 농도 저하, 지질 대사 관련 효소 및 전사인자 발현 에 미치는 영향 38
• 1) 혈중 지질농도 38
• (1) 혈중 중성지방 농도 38
• (2) 혈중 콜레스테롤 농도 38
• 2) 혈중 ALT, AST 농도 41
• 3) 간의 지질 농도 43
• 4) 간의 지질 대사 관련 효소 및 관련 전사인자의 발현 정도 46
• (1) 지질 대사 관련 효소 발현 정도 46
• ① Fatty acid synthase 발현 정도 46
• ② HMG-Co A reductase 발현 정도 48
• (2) 지질 대사 관련 핵 전사인자 발현 정도 측정 50
• ① SREBP-1 발현 정도 50
• ② SREBP-2 발현 정도 52
• Ⅳ. 요약 및 결론 54
• 참고문헌 58
• Abstract 76