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홍삼의 에탄올 추출물의 감마선 조사를 통한 항암효과 증대
허정무 ( Jung Mu Hur ),김동호 ( Dong Ho Kim ) 한국응용생명화학회 2011 Journal of Applied Biological Chemistry (J. Appl. Vol.54 No.1
Red ginseng is one of the most popular traditional medicines in Korea. In this study, we developed a new technique in which ethanol extract of red ginseng (HRG) was exposed to the Co60 gamma radiation ranging from 1~5 kGy. The irradiated HRG (IHRG) were analyzed by high performance liquid chromatography (HPLC) to determine any compositional changes of ginsenosides due to irradiation. No appreciable difference was observed in the HPLC pattern of ginsenosides of HRG. Using MTT assay, the cytotoxicity effect was significantly increased by IHRG compared to HRG. The LD50 concentration was 30 μg/mL for IHRG-1 (1 kGy), and 15 μg/mL for IHRG-5 (5 kGy). The evidences of apoptosis, such as nuclei cleavage and Annexin V staining, were observed in the human prostate cancer PC-3 cells treated with the IHRG. Additionally, the level of reactive oxygen species (ROS) was apparently elevated by IHRG. We also studied the inhibitory effect of IHRG on the growth rate of tumor xenografts in BALB/c male mice. The tumor growth rates were inhibited by 56.9 and 76.1% in mice treated with 10 mg/kg of IHRG-1 and IHRG-5, respectively, compared with control group (21.1%). These results suggest that some biologically active and soluble components in HRG can be more effectively enhancement of anti-tumor effects using irradiation.
TNBS에 의해 유도된 마우스 대장염모델에서 유산균 발효 마의 항염효과
현미선 ( Mee Sun Hyun ),허정무 ( Jung Mu Hur ) 한국응용생명화학회 2011 Journal of Applied Biological Chemistry (J. Appl. Vol.54 No.1
To develop a health-aid preparation of Dioscorea batatas (DB), lactic acid fermentation was attempted using a mixed starter comprising of Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium longum. The anaerobic fermentation of a 10% DB flour suspension gave a uniform suspension of pH 3.65, containing 8×106 CFU/mL lactic acid bacteria. During the administration of the lactic acid fermented DB (FDB) and DB to trinitrobenzene sulfonic acid (TNBS)-induced colitis mouse model, histological lesions, morphological damage, and myeloperoxidase acitivity were significantly reduced at a dosage of 200 and 400 mg/kg/day. Dose-response (200 and 400 mg/kg/day) studies revealed that FDB pre-treatment of mice significantly ameliorated the appearance of diarrhoea and the disruption of colonic architecture. In FDB-pretreated mice, there was a significant reduction in the degree of both neutrophil infiltration (measured as decrease in myeloperoxidase activity) and weight loss rates. Theses findings suggest that FDB exerts beneficial effects in experimental colitis and may be useful in the treatment of inflammatory bowel disease.
흰쥐 평활근세포에서 gliotoxin 세포독성에 대한 PKC의 효과
방현주(Hyun-Ju Bang),김진오(Jin-O Kim),오정택(Jung-Taek Oh),이한솔(Han-Sol Lee),정용관(Yong-Kwan Cheong),허정무(Jung-Mu Hur),오재민(Jay-Min Oh),최민규(Min-Kyu Choi),박승택(Seung-Taeck Park),정연태(Yeun-Tai Chung) 대한해부학회 2003 Anatomy & Cell Biology Vol.36 No.5
Gliotoxin은 Penicillium, Aspergillus, Gliocladium 및 Thermoascus와 같은 곰팡이들로부터 생성되며 여러 가지 곰팡이에 의한 질병이나 면역조절기능 같은 다양한 생리적 효과를 나타낸다. 최근에 다양한 세포주에서 gliotoxin에 의한 아폽토시스의 기전이 밝혀지고 있으며, Gliotoxin에 의한 아폽토시스 유도 시 칼슘의 농도가 증가하는 현상이 보고 되었다. 본 연구에서는 칼슘 및 칼슘과 상보적인 PKC를 전처리 한 세포에서 gliotoxin의 효과에 대하여 살펴보았다. Gliotoxin에 의한 세포의 손상에는 bcl-2 family인 bad의 발현이 증가하는 현상을 보였으며, 그와 반대로 bcl-2의 발현은 감소하였다. 또한 gliotoxin을 처리한 세포에서는 시간 의존적으로 caspase-3의 활성 증가와 DNA 분절현상이 관찰되었다. 칼슘과 PKC를 전 처리한 세포에서는 gliotoxin에 의한 세포의 손상을 억제하였으며, bcl-2의 발현이 대조군과 유사하였고 bad는 발현되지 않았다. 또한 caspase-3를 비활성화 시키고, DNA 분절을 억제시켰다. 이들 결과를 통하여 알 수 있는 것은, gliotoxin에 의한 세포의 죽음은 사립체에서 cytochrome c의 방출과 동시에 pro-apoptotic bcl-2 family의 발현을 통해서 caspase-3가 활성화되고, 그 결과 DNA 분절을 일으켜 아폽토시스를 유발시키는 것으로 추정된다. 칼슘 및 PKC를 전처리한 경우에는 사립체에서 bcl-2 family 부분에서 bad와 같은 pro-apoptotic bcl-2 family의 발현을 억제시킴으로써 그 후에 오는 caspase-3의 활성을 차단하는 것으로 사료된다. Aspergillus funigatus and other pathogenic fungi synthesize a toxic epidithi- odiopiperzine (ETP) metabolite, namely gliotoxin. Gliotoxin commonly react with sulfhydryl groups, and then, forms hydrogen peroxide. These fungal toxins induce apoptotic cell death in various cells. Apoptosis induced by gliotoxin need calcium. Effect of calcium preconditioning was not reported in gliotoxin-induced apoptosis. To examine the effect of protein kinase C (PKC) and calcium which was regulate caspase-3, PKC and calcium preconditioning before gliotoxin treatment, apoptotic agents such as bcl-2 family, caspase-3 and DNA fragmentation in A7r5 cell line from rat smooth muscle cell were studied. These results showed that gliotoxin induces the expression of bad of bcl-2 family, caspase-3 activation and DNA fragmentation in A7r5 cells. Gliotoxin treatment followed by calcium and PKC preconditioning suppress the Bad of bcl-2 family, and inhibited caspase-3 activation, respectively. These results suggest that PKC and calcium preconditioning protect the gliotoxin-induced apoptosis, through the protection of pro-apoptotic bcl-2 family in A7r5 cells.
LPS로 유도된 대식세포 활성에서 발효 산약 추출물의 억제 효과
임성원(Sung-Won Lim),이선희(Sun-Hee Lee),허정무(Jung-Mu Hur),이영미(Young-Mi Lee),김대기(Dae-Ki Kim) 대한약학회 2011 약학회지 Vol.55 No.5
This study was to elucidate the anti-inflammatory activities of a methanol extract derived from the fermented bark of Dioscoreae batatas on LPS-induced activation in macrophages. It was fermented with Lactobacillus fermentum and L. plantarum and then analyzed to identify the contents of methanol extract and diosgenin. The fermented product showed 3-fold increase in the extraction yield by methanol, and 1.8-fold increase in diosgenin contents, compared to that from the dried bark of D. batatas. Although the methanol extract from the unfermented D. batatas inhibited lipopolysaccharide (LPS)-induced production of nitric oxide (NO) and TNF-α in J774 A.1 cells, the methanol extract from the fermented product revealed significantly the enhanced the inhibitory activities on LPS-induced production of NO and TNF-α. Taken together, our results indicate that fermentation of bark of D. batatas elevates the functional activity inhibiting macrophage activation through the increase of the content of anti-inflammatory compounds. Thus, its methanol extract may be useful as a functional material for the therapy of inflammatory diseases.