인체혈구암세포에서 β-sitosterol에 의한 세포주기 S Arrest 및 XIAP 발현저하에 의한 Apoptosis 유발

박철,최병태,이원호,김기영,최영현 대한암예방학회 2007 Journal of cancer prevention Vol.12 No.2

Phytosterols are enriched in legumes, oil seeds and unrefined plant oils as found in foods such as peanut butter, pistachios and sunflower seeds. Previous results indicated that β-sitosterol, a most abundant phytosterol, induce growth arrest and apoptosis of several specific types of tumor cells in vitro and decreases the size and the extent of tumor metastases in vivo. However, the mechanism by which β- sitosterol induces anti-proliferation is not completely understood. This study investigated the mechanism of anti-proliferation induced by β-sitosterol in human leukemic U937 cells. Treatment of U937 cells to β-sitosterol resulted in the growth inhibition and morphological change in a dose-dependent manner. Flow cytometric analysis revealed that β-sitosterol treatment caused S phase arrest of the cell cycle and increased populations of apoptotic-sub G1 phase. β-sitosterol induced apoptosis through down-regulation of inhibited X-linked inhibitor of apoptosis (XIAP) expression without alteration Bcl-2 family. Z- DEVD-fmk, caspase-3 specific inhibitor, increased the survival rate of β-sitosterol-treated U937 cells. Although β-sitosterol did not alter Bcl-2 family, Bcl-2 overexpression cells blocked the decrease of XIAP expression. These results show that β-sitosterol potently induces S arrest and apoptosis in U937 cells, and β-sitosterol-induced apoptosis is related to the down-regulation of XIAP. And β-sitosterol can be used as a new therapeutic approach for the treatment of human monocytic leukemia cells. (Cancer Prev Res 12, 128-135, 2007)

Evaluation of the antiproliferative effect of β-sitosterol isolated from Combretum platypetalum Welw. ex M.A. Lawson (Combretaceae) on Jurkat-T cells and protection by glutathione

Auxillia Machingauta,Marc Y. Stevens,Chi Godloves Fru,Simbarashe Sithole,Samuel Yeboah,Stanley Mukanganyama 경희대학교 융합한의과학연구소 2022 Oriental Pharmacy and Experimental Medicine Vol.22 No.4

Combretum species are distributed widely in Southern Africa and are known for their medicinal properties. The species have potential as sources of anticancer agents. Combrestanin-A4 isolated from Combretum caffrum is one of the pure compounds now under clinical trials. The aim of this study was to fractionate and isolate plant phytoconstituents of C. platypetalum and determine their antiproliferative effects on a human leukemic cancer cell line, Jurkat-T cells. Dried powdered leaf plant samples were extracted serially with hexane, DCM, acetone, ethyl acetate, ethanol, methanol, and water. The total combined extracts were run on a silica gel column using a mobile phase of increasing polarity. β-Sitosterol was isolated from pool 94–98 of the fractions and identified by 1H-NMR and 13C-NMR and its molecular formula confirmed by mass spectrometry. The effects of β-sitosterol on proliferation of cells, effect of combining β-sitosterol and glutathione, effect of combining β-sitosterol and camptothecin and effect of β-sitosterol on glutathione S-transferase activity were determined. β-sitosterol showed dose-dependent antiproliferative effects against Jurkat-T cells and these effects were shown to be irreversible. Reduced glutathione protected the cells from the effects of β-sitosterol. Enhance antiproliferative effects were observed when β-sitosterol was combined with camptothecin. β-sitosterol was also shown to inhibit the activity of glutathione S-transferases in the cancer cell line. The results of the study suggested that β-sitosterol has antiproliferative effects in the Jurkat-T cells. Further work needs to be done on normal cells to determine if β-sitosterol affects cancer cells only.

흰깨 추출물과 β-Sitosterol이 H1299 폐암세포의 성장, 이동, 부착에 미치는 효과

이중재(Jungjae Lee),김서윤(Seoyun Kim),주지형(Jihyeung Ju) 한국식품영양과학회 2015 한국식품영양과학회지 Vol.44 No.9

본 연구에서는 흰깨의 에탄올 추출물이 폐암세포의 성장, 이동, 부착 등에 미치는 영향을 인체유래 폐암 세포주인 H1299 세포를 이용하여 in vitro 수준에서 조사하고, 이러한 흰깨 추출물의 효과를 흰깨의 주요 활성 성분 중 하나인 β-sitosterol의 효과와 비교하고자 하였다. 흰깨 추출물(150~600 μg/mL)과 β-sitosterol(3.125~25 μM)은 H1299 세포의 성장을 각각 대조구 대비 51.5~82.6%와 27.5~49.0%로 억제하는 농도 의존적 활성을 나타내었고, 이러한 흰깨 추출물과 β-sitosterol의 세포 성장 억제 활성은 부분적으로 apoptosis 유도 활성에서 기인되는 것으로 생각된다. 흰깨 추출물(150~600 μg/mL)은 H1299 세포의 이동과 부착을 억제하는 활성을 나타내지 않은 반면에 β-sitosterol 은 3.125~25 μM 농도에서 세포 이동을 대조구 대비 80.8~ 86.2%로, 6.25~25 μM 농도에서 세포 부착을 대조구 대비21.5~37.4%로 각각 억제하는 활성을 나타내었다. 이상의 연구 결과를 종합하여 볼 때 H1299 폐암세포의 성장, 이동, 부착 등을 억제하는 데에는 흰깨의 추출물보다는 단일성분인 β-sitosterol이 더 효과적인 것으로 생각된다. 앞으로 이와 같은 연구 결과가 in vivo 수준에서 재현되는지 여부를 검증하고 관련 기전을 탐색하는 것이 필요할 것으로 생각된다. The current study aimed to investigate effects of ethanol extract of white sesame seed (WSE) as well as a major constituent of white sesame seed, β-sitosterol, on the growth, migration, and adhesion of H1299 human lung cancer cells. Treatment with WSE at concentrations of 150, 300, and 600 μg/mL dose-dependently inhibited cell growth (to 51.5∼82.6% of control). Treatment with β-sitosterol at concentrations of 3.125, 6.25, 12.5, and 25 μM inhibited cell growth to a greater extent (to 27.5∼49.0% of control) than that with WSE (P<0.05). Treatment with WSE (at concentration of 600 μg/mL) or β-sitosterol (at concentration of 25 μM) resulted in increased sub-G1 cell population, indicating their apoptosis-inducing activities. β-Sitosterol was effective in inhibiting both cell migration (to 80.8∼86.2% of control at a concentration range of 3.125∼25 μM) and adhesion (to 21.5∼37.4% of control at a concentration range of 6.25∼25 μM), whereas WSE at a concentration range of 150∼600 μg/mL was ineffective. These results indicate that β-sitosterol is more active than WSE in inhibiting growth, migration, and adhesion of H1299 human lung cancer cells. Further studies are needed to determine if similar effects are reproduced in vivo.

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.

The Ameliorative Effect of β-sitosterol on DNCB-induced Atopic Dermatitis in Mice

Su-Jin Kim 대한의생명과학회 2017 Biomedical Science Letters Vol.23 No.4

β-sitosterol, one of phytosterols, exhibited numerous pharmacological effect including anti-inflammatory, anti-cancer and immune-modulating properties. This study attempted to determine the pharmacological effects of β-sitosterol on atopic dermatitis (AD). We investigated to ascertain the pharmacological effects of β-sitosterol on 2, 4-dinitrochlrobenzene (DNCB)-induced AD symptom and histamine-induced scratching behaviors in mice. Additionally, we evaluated the effects of β-sitosterol on the interleukin (IL)-6 levels in HaCaT cells and skin tissue of AD. The findings of this study demonstrated that β-sitosterol reduced AD clinical symptoms such as eczematous, erythema and dryness and serum histamine and IgE levels in DNCB-induced AD model and histamine-induced scratching behaviors in mice. Additionally, β-sitosterol inhibited the IL-6 expression in AD-like skin lesion and HaCaT cells. Collectively, these findings provide that β-sitosterol could be a therapeutic agent for skin inflammation including AD.

β-Sitosterol treatment attenuates cognitive deficits and prevents amyloid plaque deposition in amyloid protein precursor/ presenilin 1 mice

Jian-Ya Ye,Li Li,Qing-Mao Hao,Yong Qin,Chang-Sheng Ma 대한생리학회-대한약리학회 2020 The Korean Journal of Physiology & Pharmacology Vol.24 No.1

Alzheimer’s disease (AD) is the most common neurodegenerative disorder causing dementia worldwide, and is mainly characterized by aggregated β-amyloid (Aβ). Increasing evidence has shown that plant extracts have the potential to delay AD development. The plant sterol β-Sitosterol has a potential role in inhibiting the production of platelet Aβ, suggesting that it may be useful for AD prevention. In the present study, we aimed to investigate the effect and mechanism of β-Sitosterol on deficits in learning and memory in amyloid protein precursor/presenilin 1 (APP/PS1) double transgenic mice. APP/PS1 mice were treated with β-Sitosterol for four weeks, from the age of seven months. Brain Aβ metabolism was evaluated using ELISA and Western blotting. We found that β-Sitosterol treatment can improve spatial learning and recognition memory ability, and reduce plaque load in APP/PS1 mice. β-Sitosterol treatment helped reverse dendritic spine loss in APP/PS1 mice and reversed the decreased hippocampal neuron miniature excitatory postsynaptic current frequency. Our research helps to explain and support the neuroprotective effect of β-Sitosterol, which may offer a novel pharmaceutical agent for the treatment of AD. Taken together, these findings suggest that β-Sitosterol ameliorates memory and learning impairment in APP/PS1 mice and possibly decreases Aβ deposition.

β-Sitosterol이 HCT116 대장암세포의 콜로니형성 및 부착성에 미치는 효과

송보람(Bo ram Song),주지형(Ji hyeung Ju) 충북대학교 생활과학연구소 2016 생활과학연구논총 Vol.20 No.1

β-Sitosterol is the most abundant phytosterol present ubiquitously in plants. The aim of the current study was to investigate the inhibitory effects of β-sitosterol on colony formation and adhesion in HCT116 human colon cancer cells. β-Sitosterol significantly inhibited cell growth (to 50-89% of the control by the concentrations of 25-100 μM; p<0.01) and effectively changed nuclear morphology (by the concentration of 50 μM) in HCT116 cells. These results confirm previous findings on the growth-inhibitory and apoptosis-inducing activities of β-sitosterol in different cancer cell lines. Treatment with β-sitosterol resulted in significant inhibition of both colony formation (to 29% of the control by the concentration of 50 μM) and adhesion (to 44-54% of the control by the concentration of 25-100 μM) in HCT116 cells. These results suggest that β -sitosterol might modulate the self-renewal capacity of colon cancer stem cells as well as invasion and metastasis during colon carcinogenesis. More in-depth studies are needed in order to verify such actions and to elucidate underlying mechanisms.

미백, 보습 및 탈모방지에 대한 황칠나무(Dendropanax modifera Lev.)에서 분리한 1-tetradecanol, β-sitosterol의 효과

이선영 ( Sun Young Lee ),최은진 ( Eun-jin Choi ),배동혁 ( Dong-hyuck Bae ),이동욱 ( Dong-wook Lee ),김선오 ( Sunoh Kim ) 대한화장품학회 2015 대한화장품학회지 Vol.41 No.1

황칠나무(Dendropanax mobifera Lev.)는 두릅나무과에 속하는 난대성 수종으로 대한민국 남쪽지방에 주로 분포하며 전통적인 민간요법 약재로 사용되어 왔으나 지금까지 피부에서의 효능이 잘 알려지지 않았다. 우리는 본 연구에서 황칠나무 잎의 열수추출물과 n-hexane 분획물에서 1-tetradecanol과 β-sitosterol 성분을 분리정제하여 다양한 피부개선효과와 탈모방지효과를 검증하였다. 1-tetradecanol의 타이로시나아제 활성 저해 효과는 열수추출물과 n-hexane분획물 보다 높은 저해 활성이 나타났다. 그리고 B16F10 세포를 이용한 멜라닌 양을 측정한 결과 1-tetradecanol은 세포독성 없이 농도 의존적으로 멜라닌 양이 감소됨을 확인하였다. 실험동물을 이용한 경표피수분손실량(TEWL)에서도 1-tetradecanol에서 수분 손실이 가장 낮게 나타났다. β-sitosterol의 탈모 방지에 대한 효과는 HR-1 hairless mice의 성장 주기별 특성을 이용하여 관찰하였다. 그 결과 β-sitosterol을 처리한 마우스의 탈모는 지연되었고 모발의 밀도도 가장 높게 나타났다. 그러므로 황칠 n-hexane분획물에서 분리한 1-tetradecanol과 β-sitosterol이 미백과 보습 용도의 화장품 기능성 원료 및 탈모 개선에 가능성이 있는 화장품소재로서 상업적 발전의 가능성을 보여주고 있다. Dendropanax morbifera Leveille (Araliaceae) is an endemic species growing in the south-western part of South Korea and has been used in folk medicine. However, the effects of Dendropanax morbifera Lev. on skin biology remain to be elucidated. In this study, we isolated 1-tetradecanol and β-sitosterol from the n-hexane fraction of Dendropanax mobifera Lev. and To investigate the whitening effect of the fraction, we tested the inhibition of tyrosinase activity of 1-tetradecanol. The results show that the inhibitory effect of the 1-tetradecanol was higher than water extract and n-hexane fraction. And 1-tetradecanol significantly reduced melanin contents of B16F10 cells compared to more than water extract and n-haxane fraction dose-dependantly without cell cytotoxicitiy (below 100 μg/mL). We also investigated the skin moisturizing effect using HR-1 hairless mice. The transepidermal water loss (TEWL) in the 1-tetradecanol treated group was significantly smaller than that in the other groups. To investigate the effect of the preventing hair loss by β-sitosterol, we observed HR-1 hairless mice through periodic growth feature. The results suggest that hair loss of mice by β-sitosterol was delayed and it’s hair density showed the highest. These data provide evidence that Dendropanax morbifera Lev. may be a potent candidate for the improvement of both skin whitening, moisturizing and alopecia from the point of cosmetic industry view.

Schima wallichii sp. liukiuensis로 부터 분리된 β-Sitosterol Glycoside 항균물질의 안정성 및 돌연변이원성

최명석,신금,이동권,권오웅,손성호 성균관대학교 약학연구소 1999 成均藥硏論文集 Vol.11 No.-

Abstract-Stability of the β-sitosterol glycoside from Schima wallichii sp. liukiuensis at various physical conditions were investigated, mutagenecity of the steroid saponin was determined by Ames test. When exposed in pH 3 to pH 8, the β-sitosterol glycoside was stable on antimicrobial activity against yeasts. The antimicrobial activity of the β-sitosterol glycoside also stable in high temperature, N_2, O_2 gas and light exposure, and metal ion. Ames test result revealed thatβ-sitosterol glycoside did not have any mutagenic activity. These results suggest that the β-sitosterol glycoside might be a promising condidate as a natural antimicrobial compound.

Induction of NQO1 by Plant Sterols in Murine Hepatoma Cells

Ji Yeon Seo,Hee Jung Kang,Woo-Keun Kim,Jong-Sang Kim 대한암예방학회 2007 Journal of cancer prevention Vol.12 No.4

Our preliminary study showed that methanolic extract of Chrysanthemum coronarium had potential to induce detoxifying enzymes such as NAD(P)H:Quinone Oxidoreductase 1 (NQO1) and glutathione S-transferase (GST) activity and β-sitosterol was identified to be one of major detoxifying enzyme inducers present in the vegetable. In this study we attempted to evaluate the potential to induce detoxifying enzymes of plant sterols which share structural similarity with β-sitosterol. NQO1 activity was significantly induced by campesterol and β-sitosterol but not by ergosterol and stigmaterol in hepa1c1c7 cells. Campesterol and β-sitosterols also increased the expression of γ-glutamylcysteine synthase but not glutathione reductase. However, all plant sterols used in the study failed to induce ARE-luciferase activity in HepG2-C8 cells, suggesting NQO1-inducing abilities of β-sitosterol and campesterol might be independent of ARE-mediated transcriptional activation of detoxifying genes. (Cancer Prev Res 12, 268-272, 2007) Key Words: Plant sterols, Campesterol, β-sitosterol, Detoxifying enzymes