The present study investigated an ethanol extract of Chaenomeles sinensis fruit (CSF) for possible neuroprotective effects on neurotoxicity induced by amyloid β protein (Aβ) (25-35) in cultured rat cortical neurons and also for antidementia activity in mice. Exposure of cultured cortical neurons to 10μM Aβ (25-35) for 36 h induced neuronal apoptotic death. At 0.1-10μg/ml, CSF inhibited neuronal death, elevation of intracellular calcium concentration ([Ca2+]i), and generation of reactive oxygen species (ROS) induced by Aβ (25-35) in primary cultures of rat cortical neurons. Memory loss induced by intracerebroventricular injection of mice with 15 nmol Aβ (25-35) was inhibited by chronic treatment with CSF (10, 25 and 50 mg/kg, p.o. for 7 days) as measured by a passive avoidance test. CSF (50 mg/kg) inhibited the increase of cholinesterase activity in Aβ (25-35)-injected mice brain. From these results, we suggest that the antidementia effect of CSF is due to its neuroprotective effect against Aβ (25-35)-induced neurotoxicity and that CSF may have a therapeutic role for preventing the progression of Alzheimer's disease.
Vitis amurensis (VA; Vitaceae) has long been used in oriental herbal medicine. It has been reported that roots and seeds of VA have anti-inflammatory and antioxidant effects. In the present study, the protective effect of ethanol extract from stems and leaves of VA on hydrogen peroxide (H2O2) (100 μm)-induced neuronal cell damage was examined in primary cultured rat cortical neurons. VA (10-100 μg/ml) concentration-dependently inhibited H2O2-induced apoptotic neuronal cell death measured by 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and Hoechst 33342 staining. VA inhibited H2O2-induced elevation of intracellular Ca2+ concentration ([Ca2+]i) and generation of reactive oxygen species (ROS), which were measured by fluorescent dyes. Pretreatment of VA also prevented glutamate release into medium induced by 100 μm H2O2, which was measured by HPLC. These results suggest that VA showed a neuroprotective effect on H2O2-induced neuronal cell death by interfering with H2O2-induced elevation of [Ca2+]i, glutamate release, and ROS generation. This has a significant meaning of finding a new pharmacological activity of stems and leaves of VA in the CNS.
An advanced extraction method by ultrasonic extraction with applied solid phase extraction (SPE) has been developed for the determination of simultaneous eight major ginsenosides, namely ginsenosides Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, and Rd in the root of Panax ginseng. Four extraction methods including n-BuOH reflux extraction (Method A), 70% EtOH reflux extraction (Method B), 50% MeOH reflux extraction with SPE (Method C), and 50% MeOH ultrasonication with SPE clean-up process (Method D) were investigated for the determination of eight major ginsenosides. Total contents of ginsenosides were highest by extraction of Method C as 2.408±0.011%. However, Method D was evaluated as relatively simpler and more efficient method due to short extraction time, small solvent consumption and less expensive, compared to conservative reflux method. Ginsenosides were also satisfactorily separated with good resolution and the accuracy range was between 1.05 and 4.06% as relative standard deviation (RSD) by Method D. SPE condition and HPLC condition were further optimized for determination of eight major ginsenosides by the ultrasonic extraction method. Conclusively, ultrasonic extraction of 2 g sample of ginseng using ultrasonic bath and 1 loading for SPE was evaluated as proper condition for extraction of ginseng.
Moutan cortex, the root bark of Paeonia suffruticosa Andrews (Paeoniaceae), has pharmacological effects such as anti-inflammatory, antiallergic, analgesic and antioxidant activities. We investigated a methanol extract of Moutan cortex for neuroprotective effects on neurotoxicity induced by amyloid β protein (Aβ) (25-35) in cultured rat cortical neurons. Exposure of cultured cortical neurons to 10 μM Aβ (25-35) for 24 h induced neuronal apoptotic death. Moutan cortex inhibited 10 μM Aβ (25-35)-induced neuronal cell death at 30 and 50 μg/ml, which was measured by a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and Hoechst 33342 staining. Moutan cortex inhibited 10 μM Aβ (25-35)-induced elevation of intracellular calcium concentration ([Ca2+]i), and generation of reactive oxygen species (ROS) which were measured by fluorescent dyes. Moutan cortex also inhibited glutamate release into medium induced by 10 μM Aβ (25-35), which was measured by HPLC. These results suggest that Moutan cortex prevents Aβ (25-35)-induced neuronal cell damage by interfering with the increase of [Ca2+]i, and then inhibiting glutamate release and ROS generation. Moutan cortex may have a therapeutic role in preventing the progression of Alzheimer's disease.
본 시험는 국내의 리그난 고함유 품종 개발이 미흡한 실정에서, 기존에 개발된 참깨 품종을 대상으로 항산화 성분인 리그난 화합물 및 지방산 조성 등의 품질을 분석 비교함으로써 리그난 고함유 품종 개발과 리그난 화합물의 건강기능식품으로의 소재 개발에 유용하게 이용될 수 있는 품질 기초 자료를 제시하고자 실시하였으며 그 결과를 요약하면 다음과 같다. 1. 2002년, 2003년에 재배된 참깨 각각 21품종, 27품종을 대상으로, 2002년도의 세사민, 세사몰린의 평균함량은 각각 4.08±1.742.48±0.68mg/g 이었으며, 2003년도의 세사민, 세사몰린의 평균함량은 각각 4.08±1.512.48±0.67mg/g 이었다. 2. 흰깨, 검은깨의 2003년의 세사민 평균함량은 4.87±0.731.82±0.47mg/g , 세사몰린 함량은 2.85±0.251.42±0.31mg/g 으로 세사민, 세사몰린 모두 흰깨가 검은깨보다 함량이 2~~4 배정도 높았다. 3. 성분깨는 세사민, 세사몰린 함량이 2002년에는 각각 5.94, 3.26mg/g이었고, 2003년에는 각각 6.27, 3.21mg/g으로 모두 상위 1~~2 위 순위에 분포하는 양상이었으나, 다른 흰깨 품종들은 재배 연수에 따라 절대함량은 물론 함량 순위에도 약간의 변이를 보였다. 4. 세사민은 2002년에는 단백깨(6.22mg/g), 성분깨(5.94mg/g), 남다깨(5.83 mg/g), 남안깨(5.59 mg/g) 순으로 함량 순위를 나타내었고, 2003년에는 성분깨(6.27 mg/g), 다삭깨(5.53mg/g), 단백깨(5.50mg/g), 진백깨(5.45 mg/g), 서둔깨(5.41mg/g) 순으로 함량 순위를 나타내었다. 5. 세사몰린은 2002년에는 황백깨(3.27mg/g), 성분깨(3.26mg/g), 안남깨(3.22 mg/g), 한섬깨(3.17 mg/g), 단백깨(3.16mg/g)순으로, 2003년에는 성분깨(3.21 mg/g), 서둔깨(3.04 mg/g), 풍안깨(3.10 mg/g), 남안깨(3.09 mg/g), 단백깨(3.04 mg/g) 순으로 함량 순위를 나타내었다. 6. 세사민 함량이 높은 단백깨, 성분깨, 다삭깨, 남다깨, 풍안깨 등은 성분깨가 48.56~% 로서 기름함량이 다소 낮았으나 나머지는 50.00~% 이상으로 양호한 기름 함량분포를 나타내었다.
21 종의 생약 추출물을 선정하여 5 μg/ml의 농도에서 SK-MEL-28 cell 에 대한 세포독성을 조사한 결과 목단피의 메탄올 추출물이 74.3%의 성장율을 보였다. 활성 물질을 찾기 위하여 목단피로부터 silica gel column chromatography를 실시하여 hexane 분획과 EtOAc 분획에서 충 5개의 화합물을 분리하였고 mp, UV, IR, 1H-NMR, 13C-NMR 등 각종 물리, 화학적 data로부터 그 구조를 paeonol (1), benzoylpaeoniflorin (2), benzoic acid (3), 2,5-dihydroxy-4-methoxy-acetophenone (4), paeoniflorin (5)으로 동정하였다. 분리한 물질을 human 피부 암세포인 SK-MEL-28 세포주에 대하여 10 μg/ml의 농도에서 SRB방법으로 세포독성을 측정한 결과 compound 4가 ED50 값이 5.92 μg/ml로 가장 좋은 세포독성을 나타내었다. 이결과는 compound 4가 SK-MEL-28 melanoma 세포주에 대한 새로운 항암 후보 물질임을 제시한다.
200종의 식물추출물을 대상으로 PEP 저해활성을 측정한 결과, 81종의 식물추출물은 10~30% 범위의 저해율을, 28종의 식물추출물은 10%이하의 저해율을 나타내어 총 200종의 식물추출물 중 약 55% 정도의 식물추출물이 30%이하의 낮은 PEP 저해활성을 나타내었다. 반면에, Prunus mume (매화나무) 잎 (95.3%), 노루발과 (Pyrolaceae)의 Pyrola japonica (노루발) 잎과 줄기 (93.2%), 물레나물과 (Hypericaseae)의 Hypericum ascyron (물레나물) 지상부 (90.2%), 범의귀과(Saxifragaceae)의 Astilbe chinensis var. typica (노루오줌) 지상부 (90.1%), 보리수나무과 (Elaeagnaceae)의 Elaeagnus umbellata (보리수나무) 잎과 줄기 (90.1%)등 5종의 추출물들이 5 ppm에서 PEP에 대하여 90% 이상의 강한 저해활성을 나타내었다.