The ameliorating effects of stigmasterol on scopolamine-induced memory impairments in mice

Park, Se Jin,Kim, Dong Hyun,Jung, Jun Man,Kim, Jong Min,Cai, Mudan,Liu, Xiaotong,Hong, Jin Gyu,Lee, Chang Hwan,Lee, Kang Ro,Ryu, Jong Hoon Elsevier 2012 european journal of pharmacology Vol.676 No.1

<P><B>Abstract</B></P><P>Stigmasterol, a kind of phytosterol, is present in small amounts in various foods. In the present study, we investigated the effects of stigmasterol on scopolamine-induced memory impairments using the passive avoidance and the Morris water maze tasks in mice. In addition, changes in memory-related molecules, including extracellular signal-regulated kinase (ERK) and cAMP response element-binding protein (CREB), were examined following the administration of stigmasterol. Scopolamine-induced memory impairments were significantly attenuated by the administration of stigmasterol (10mg/kg) in the passive avoidance task. In the Morris water maze task, the escape latencies were significantly decreased in the stigmasterol-treated group compared to the scopolamine-treated group during the training phase. The swimming times within the target zone during the probe trial were significantly increased as compared to scopolamine-treated mice. Furthermore, the ameliorating effect of stigmasterol on scopolamine-induced memory dysfunction was blocked by a sub-effective dose of dizocilpine (MK-801), an NMDA receptor antagonist, and tamoxifen, an estrogen receptor antagonist, in the passive avoidance task. In addition, the expression levels of phosphorylated ERK and CREB in the hippocampus were significantly increased by stigmasterol, which was blocked by tamoxifen or MK-801 with scopolamine. These results suggest that stigmasterol-induced cognitive ameliorative effects are mediated by the enhancement of cholinergic neurotransmission system via the activation of estrogen or NMDA receptors.</P>

Stigmasterol 유도체의 합성 및 이들의 아토피 억제 효과

이지현,조수현,신영진,송규용 충남대학교 약학대학 의약품개발연구소 2012 藥學論文集 Vol.27 No.-

The house dust mite(D. pteronissinus, Dp) plays a key role in the pathogenesis of allergic diseases, including atopic dermatitis and asthma. Monocyte chemoattractant protein 1(MCP-1), interleukin-6 (IL-6) and interleukin-8 (IL-8) play a pivotal role in mediating the infiltration of various cells into the skin of atopic dermatitis. We have attempted to synthesize various stigmasterol derivatives, especially modified cinnamoyl stigmasterol. We found that S14 (2-fluorocinnamoyl stigmasterol), S15(3-fluorocinnamoyl stigmasterol) potently suppressed the elevated production of IL-6 and IL-8 induced by D. pteronissinus extract (DpE) treatment in the human monocytic cell line, THP-1 cells. But all modified cinnamoyl stigmasterol don't suppressed the elevated production MCP-1 induced by DpE treatment in THP-1 cell. Taken together, S14 and S15 may be useful for research on atopic dermatitis and new lead compounds for atopic dermatitis.

애기마름(Trapa pseudoincisa S. et. Z.)으로부터 sterol 화합물의 분리 및 구조동정

양혜정,송명종,노영덕,김대근,백남인 경희대학교식량자원개발연구소 2007 硏究論文集 Vol.26 No.1

애기마름을 80% MeOH로 추출하고, 얻어진 추출물을 EtOAc, n-BuOH 및 H_(2)O로 용매 분획 하였다. EtOAc 분획으로부터 silica gel과 octadecyl silica gel(ODS) column chromatography를 반복 실시하여 2종의 화합물을 분리하였다. 각 화합물의 화학 구조는 NMR, MS 및 IR 등의 스펙트럼을 데이터를 해석하여, stigmasterol과 stigmasterol 3-O-β-D-glucopyranosyl로 구조 동정하였다. 이화합물은 애기마름에서는 처음 분리되었다. Trapa pseudoincisa was extracted with 80% aqueous MeOH, and the concentrated extract was partitioned with EtOAc, n-BuOH and H_(2)O. Two compounds were isolated from the EtOAc fraction through repeated silica gel and ODS column chromatographies. From the result of spectral data including NMR, MS, IR and GC/MS the chemical structures of the compounds were determined as stigmasterol (compound Ⅰ), and stigmasterol 3-O-β-D-glucopyranosyl (compound Ⅱ). They were isolated from Trapa pseudoincisa for the first time.

Functional Expression of Arabidopsis thaliana Sterol Glycosyltransferase from Stably Transformed Drosophila melanogaster S2 Cells

정하영,황보전,김성기,백남인,이윤형,정인식,박종화 한국생물공학회 2011 Biotechnology and Bioprocess Engineering Vol.16 No.4

Arabidopsis thaliana sterol glycosyltransferase (SGT), UGT80A2, was expressed from stably transformed Drosophila melanogaster Schneider 2 (S2) cells. Recombinant SGT was detected in both intracellular and extracellular fractions with a molecular mass of approximately 76 kDa. Secreted recombinant SGT accounted for approximately 60% of the total recombinant SGT production. Recombinant SGT in the extracellular fractions was purified to homogeneity using a simple one-step Ni-NTA affinity fractionation. Radiometrical assay using uridine diphospho-D-[U-^(14)C]glucose (UDP-^(14)C-glucose) as a sugar donor and sterols, β-sitosterol and stigmasterol, as sugar acceptors showed that the purified recombinant SGT contained UDPglycosyltransferase activity and could attach ^(14)C-glucose to β-sitosterol and stigmasterol. Recombinant SGT contained higher catalytic activity with β-sitosterol, which was similar to the recombinant SGT produced by a bacterial expression system. The transfer of ^(14)C-glucose by recombinant SGT was further determined by gas chromatography-mass spectrometry (GC-MS) analysis of cellulase-treated ^(14)C-glucosetransferred β-sitosterol and stigmasterol reactants.

리포솜 이중층의 stigmasterol이 포집된 ascorbic acid의 안정성에 미치는 영향

이동욱(Dong-Uk Lee),박혜원(Hye-Won Park),이승철(Seung-Cheol Lee) 한국식품과학회 2020 한국식품과학회지 Vol.52 No.2

SS은 식물성 스테롤로서 인체에 유익한 생리활성을 나타낸다. 리포솜 인지질을 PC과 SS로 구성하고, ASA를 탈수/재수화 방법에 의해 포집시킨 후, ASA의 안정성을 분석하였다. 리포솜의 평균 크기는 SS 함량이 증가할수록 증가하였으며, SS는 포집된 ASA의 안정성을 향상시켰다. 예를 들어, 완충용액에 존재하는 ASA는 4℃에서 8일간 저장하면 7.66%가 잔존하지만, 100:0, 90:10, 70:30 (PC/SS)로 조성된 리포솜에 포집된 ASA는 같은 조건에서 각각 34.12, 49.88, 58.58%가 잔존하였다. 이러한 결과는 리포솜에서 SS가 포집된 ASA의 안정성을 향상시킴을 의미한다. The effect of stigmasterol (SS), a phytosterol, in the liposome bilayer on the stability of encapsulated ascorbic acid (ASA) was evaluated. Liposomes, consisting of phosphatidylcholine (PC) and SS, and ASA were encapsulated by the dehydration/rehydration method. The average particle size of the liposome increased with increasing SS content. SS significantly increased the stability of encapsulated ASA. For example, ASA remaining in the liposomes of 100:0, 90:10, and 70:30 (PC:SS, w/w) ratios was 34.12%, 49.88%, and 58.58%, respectively, after storage for 8 days at 4℃, while only 7.66% ASA remained in the buffer under the same conditions. These results indicated that SS in liposomes increased the stability of encapsulated ASA.

토양 미생물로부터 생산된 Extracellular Cholesterol Oxidase의 특성

박정수(Jeong-Su Park),정종문(Jong-Moon Jeong) 한국식품영양과학회 2008 한국식품영양과학회지 Vol.37 No.11

본 연구에서는 산업적으로 사용될 수 있는 안정하고 활성이 높은 cholesterol oxidase를 생산하는 균주를 얻기 위해 토양 미생물로부터 균주를 선별하였다. 선별된 균주에 대하여 세포외 효소 활성을 측정한 결과 BEN 115로 명명한 균주가 가장 높은 효소 활성도를 나타내었다. 이 균주는 형태학적, 생리학적 특성과 배양형태 및 G+C 함량을 분석한 결과 Nocardia속으로 확인되었다. 최적 효소 생산 조건을 조사한 결과 기존 yeast malt extract broth 조성인 0.4% yeast extract, 0.4% glucose, 1% malt extract에서 가장 높은 활성을 나타냈다. 본 균주에서 생산된 extracellular cholesterol oxidase는 SDS-PAGE와 Western blot 결과 분자량이 55, 57 kDa인 두 종류의 효소가 존재하는 것으로 나타났다. BEN 115에 대한 효소학적 특성을 연구한 결과 온도 안정성은 55℃까지 효소 활성이 유지되었고, pH 안정성은 pH 3.5~9.5의 범위까지 안정한 것으로 나타났으며 최적 온도와 최적 pH는 각각 35oC와 pH 5.5인 것으로 나타났다. 또한 detergent(Triton X-100, Triton X-114 그리고 Tween 80) 첨가 시 효소 활성이 첨가하지 않은 대조군보다 약 1.6~2.0배 증가하는 것으로 나타났다. Campesterol, sitosterol 그리고 stigmasterol에 대한 기질 특이성은 cholesterol(100%)과 상대적으로 비교 시 각각 50%, 50% 그리고 27%의 기질 특이성을 나타내었다. Cholesterol oxidase catalyses the conversion of cholesterol to 4-cholesten-3-one. This enzyme has been used for clinical assay of human serum cholesterol and for reduction of cholesterol level in foods and feeds. In order to search the microorganism which has a high extracellular and stable activity of cholesterol oxidase, soil microorganisms were screened. As a result, the one with the highest extracellular cholesterol oxidase activity was obtained and named as the BEN 115. The BEN 115 strain was identified as one of the Nocardia species based on our taxonomic studies. The cholesterol oxidase from this strain was shown to have two bands of extracellular proteins on SDS-PAGE and Western blot. Their molecular masses were estimated to be about 55 and 57 kDa, respectively. In addition, this cholesterol oxidase was considerably stable at the broad range of pH 3.5~9.5 and at the temperature of 25~55℃. The optimum pH and temperature of this cholesterol oxidase were pH 5.5 and 35℃, respectively. The activity of extracellular cholesterol oxidase could be enhanced 1.6 to 2.0 folds by the addition of nonionic detergent such as Triton X-114, Triton X-100, or Tween-80 into the culturing broth. The substrate specificities against campesterol, sitosterol and stigmasterol were measured to be 50%, 50%, and 27%, respectively, compared to the cholesterol. These results suggest that Nocardia sp. BEN 115 may be useful as a microbial source of cholesterol oxidase production.

동부로부터 sterol의 분리 동정

최은희,박희정,吴倩,정인식,김지영,백남인 한국응용생명화학회 2010 Journal of Applied Biological Chemistry (J. Appl. Vol.53 No.2

The seed of cowpea (Vigna sinensis K.) was extracted with 80% aqueous methanol (MeOH). And the concentrated extract was partitioned with ethyl acetate (EtOAc), n-butanol (n-BuOH) and H2O, successively. The repeated silica gel and octadecyl silica gel (ODS) column chromatographic separations for the EtOAc and n-BuOH fractions led to isolation of four sterols. And the chemical structures of the compounds were determined as a mixture of stigmasterol and β-sitosterol with the ratio of 4 to 3 (1), 7-ketositosterol (2), and stigmasterol 3-O-β-D-glucopyranoside (3) from the interpretation of spectroscopic data including nuclear magnetic resonance (NMR) spectrum metric, mass (MS) spectrum metric and infrared (IR) spectroscope. This study reports the first isolation of β-sitosterol, 7-ketositosterol, and stigmasterol 3-O-β-D-glucopyranoside from the seed of Vigna sinensis K. In addition, compound 2, 7-ketositosterol, is rarely occurred in natural source including plant. 동부를 80% MeOH로 추출하고, 얻어진 추출물을 EtOAc, n-BuOH 및 H2O로 용매 분획 하였다. 이 중 EtOAc과 n-BuOH 분획으로부터 silica gel과 octadecyl silica gel(ODS) columnchromatography를 반복하여 4종의 sterol 화합물을 분리, 정제하였다. 각 화합물의 화학구조는 NMR, MS 및 IR 등의 spectrum data를 해석하여, stigmasterol(1a), β-sitosterol(1b), 7-ketositosterol(2) 및 stigmasterol 3-O-β-D-glucopyranoside(3)로 동정하였다. 화합물 1b, 2와 3은 동부에서 처음 분리되었다.

Evaluation of antitumor activity of stigmasterol, a constituent isolated from $Bacopa$ $monnieri$ Linn aerial parts against Ehrlich Ascites Carcinoma in mice

Ghosh, Tirtha,Maity, Tapan Kumar,Singh, Jagadish 경희한의학연구센터 2011 Oriental Pharmacy and Experimental Medicine Vol.11 No.1

Cancer remains as one of the most common cause of mortality worldwide. Cancer chemoprevention by natural products is well accepted nowadays. Phytoserols are natural products, showing anticancer activity, besides other activities. The purpose of this study was to examine the antitumor and antioxidant activity of stigmasterol, a phytosterol isolated from aerial parts of $Bacopa$ $monnieri$ Linn. against Ehrlich Ascites Carcinoma (EAC) in swiss albino mice. The effect of stigmasterol on the growth of transplantable murine tumor, life span of EAC bearing hosts, simultaneous alterations in the hematological profile, liver biochemical parameters (lipid peroxidation, antioxidant enzymes) and histopthological studies of liver were examined. Stigmasterol decreased tumor volume, packed cell volume and viable cell count, and increased mean survival time thereby increasing life span of EAC tumor bearing mice. Hematological profile reverted to near normal levels in stigmasterol treated mice. Stigmasterol decreased the levels of lipid peroxidation and increased the levels of glutathione, superoxide dismutase and catalase in the liver of EAC bearing mice. Histopathological study of liver tissues showed that stigmasterol had significant protective effect against EAC bearing mice, which corroborates the above findings. Lactate dehydrogenase (LDH) activity in ascitic fluid increased while membrane microviscosity of the EAC cells decreased upon stigmasterol treatment indicating its effect in the membrane integrity functions. The antitumor activity of stigmasterol might be mediated through the activation of protein phosphatase 2A by ceramide causing apoptosis, as is shown by structurally similar phytosterol.

Evaluation of antitumor activity of stigmasterol, a constituent isolated from Bacopa monnieri Linn aerial parts against Ehrlich Ascites Carcinoma in mice

Tirtha Ghosh,Tapan Kumar Maity,Jagadish Singh 경희대학교 융합한의과학연구소 2011 Oriental Pharmacy and Experimental Medicine Vol.11 No.1

Cancer remains as one of the most common cause of mortality worldwide. Cancer chemoprevention by natural products is well accepted nowadays. Phytoserols are natural products, showing anticancer activity, besides other activities. The purpose of this study was to examine the antitumor and antioxidant activity of stigmasterol, a phytosterol isolated from aerial parts of Bacopa monnieri Linn. against Ehrlich Ascites Carcinoma (EAC) in swiss albino mice. The effect of stigmasterol on the growth of transplantable murine tumor, life span of EAC bearing hosts, simultaneous alterations in the hematological profile,liver biochemical parameters (lipid peroxidation, antioxidant enzymes) and histopthological studies of liver were examined. Stigmasterol decreased tumor volume, packed cell volume and viable cell count, and increased mean survival time thereby increasing life span of EAC tumor bearing mice. Hematological profile reverted to near normal levels in stigmasterol treated mice. Stigmasterol decreased the levels of lipid peroxidation and increased the levels of glutathione, superoxide dismutase and catalase in the liver of EAC bearing mice. Histopathological study of liver tissues showed that stigmasterol had significant protective effect against EAC bearing mice, which corroborates the above findings. Lactate dehydrogenase (LDH) activity in ascitic fluid increased while membrane microviscosity of the EAC cells decreased upon stigmasterol treatment indicating its effect in the membrane integrity functions. The antitumor activity of stigmasterol might be mediated through the activation of protein phosphatase 2A by ceramide causing apoptosis, as is shown by structurally similar phytosterol.

스파티필름(Spathiphyllum cannifolium) 지상부로부터 지질화합물의 분리

이대영,박지해,유종수,송명종,백남인,이윤형 한국응용생명화학회 2008 Journal of Applied Biological Chemistry (J. Appl. Vol.51 No.1

The aerial parts of Spathiphyllum cannifolium were extracted with 80% aqueous MeOH and the concentrated extract was partitioned with EtOAc, n-BuOH and H2O, successively. From the EtOAc fraction, three compounds were isolated through the repeated silica gel, ODS, Sephadex LH-20 column chromatographies. From the results of physico-chemical data including NMR, MS and IR, the chemical structures of the compounds were determined as stigmasterol (1), (2S)1-O-linolenoyl-2-Omyristoyl- 3-O-β-D-galactopyranosyl-sn-glycerol (2) and stigmasterol glucoside (3). They were the first to be isolated from Spathiphyllum cannifolium. The aerial parts of Spathiphyllum cannifolium were extracted with 80% aqueous MeOH and the concentrated extract was partitioned with EtOAc, n-BuOH and H2O, successively. From the EtOAc fraction, three compounds were isolated through the repeated silica gel, ODS, Sephadex LH-20 column chromatographies. From the results of physico-chemical data including NMR, MS and IR, the chemical structures of the compounds were determined as stigmasterol (1), (2S)1-O-linolenoyl-2-Omyristoyl- 3-O-β-D-galactopyranosyl-sn-glycerol (2) and stigmasterol glucoside (3). They were the first to be isolated from Spathiphyllum cannifolium.