• BMB Reports
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• 제40권3호
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• pp.382-395
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• 2007

The herb, Cajanus indicus L, is well known for its hepatoprotective action. A 43 kD protein has been isolated, purified and partially sequenced from the leaves of this herb. A number of in vivo and in vitro studies carried out in our laboratory suggest that this protein might be a major component responsible for the hepatoprotective action of the herb. Our successive studies have been designed to evaluate the potential efficacy of this protein in protecting the hepatic as well as renal tissues from the sodium fluoride (NaF) induced oxidative stress. The experimental groups of mice were exposed to NaF at a dose of 600 ppm through drinking water for one week. This exposure significantly altered the activities of the antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), glutathione reductase (GR) and the cellular metabolites such as reduced glutathione (GSH), oxidized glutathione (GSSG), total thiols, lipid peroxidation end products in liver and kidney compared to the normal mice. Intraperitoneal administration of the protein at a dose of 2 mg/kg body weight for seven days followed by NaF treatment (600 ppm for next seven days) normalized the activities of the hepato-renal antioxidant enzymes, the level of cellular metabolites and lipid peroxidation end products. Post treatment with the protein for four days showed that it could help recovering the damages after NaF administration. Time-course study suggests that the protein could stimulate the recovery of both the organs faster than natural process. Effects of a known antioxidant, vitamin E, and a non-relevant protein, bovine serum albumin (BSA) have been included in the study to validate the experimental data. Combining all, result suggests that NaF could induce severe oxidative stress both in the liver and kidney tissues in mice and the protein possessed the ability to attenuate that hepato-renal toxic effect of NaF probably via its antioxidant activity.

• Journal of Microbiology and Biotechnology
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• 제30권11호
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• pp.1640-1650
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• 2020

Colorectal cancer (CRC) is the leading cause of common malignant neoplasm worldwide. Many studies have analyzed compositions of gut microbiota associated with various diseases such as inflammatory bowel diseases (IBD) and colon cancer. One of the most representative bacteria involved in CRC is enterotoxigenic Bacteroides fragilis (ETBF), a species belonging to phylum Bacteroidetes. We used ETBF colonized mice with azoxymethane (AOM)/dextran sulphate sodium (DSS) and zerumbone, a compound with anti-bacterial effect, to determine whether zerumbone could restore intestinal microbiota composition. Four experimental groups of mice were used: sham, ETBF colonized AOM/DSS group, ETBF colonized AOM/DSS group zerumbone 60 mg kg-1 (ETBF/AOM/DSS + Z (60)), and only zerumbone (60 mg kg^-1)-treated group. We performed reversible dye terminators-based analysis of 16S rRNA gene region V3-V4 for group comparison. Microbiota compositions of ETBF/AOM/DSS + Z (60) group and ETBF colonized AOM/DSS group not given zerumbone were significantly different. There were more Bacteroides in ETBF/AOM/DSS + Z (60) group than those in ETBF colonized AOM/DSS group, suggesting that B. fragilis could be a normal flora activated by zerumbone. In addition, based on linear discriminant analysis of effect size (LEfSe) analysis, microbial diversity decreased significantly in the ETBF colonized AOM/DSS group. However, after given zerumbone, the taxonomic relative abundance was increased. These findings suggest that zerumbone not only influenced the microbial diversity and richness, but also could be helpful for enhancing the balance of gut microbial composition. In this work, we demonstrate that zerumbone could restore the composition of intestinal microbiota.

• Nutrition Research and Practice
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• 제13권6호
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• pp.473-479
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• 2019

BACKGROUND/OBJECTIVES: Anti-inflammatory and antioxidative activities of luteolin and luteolin-7-O-glucoside were compared in galactosamine (GalN)/lipopolysaccharide (LPS)-induced hepatitic ICR mice. MATERIALS/METHODS: Male ICR mice (6 weeks old) were divided into 4 groups: normal control, GalN/LPS, luteolin, and luteolin-7-O-glucoside groups. The latter two groups were administered luteolin or luteolin-7-O-glucoside (50 mg/kg BW) daily by gavage for 3 weeks after which hepatitis was induced by intraperitoneal injection of GalN and LPS (1 g/kg BW and $10{\mu}g/kg\;BW$, respectively). RESULTS: GalN/LPS produced acute hepatic injury by a sharp increase in serum AST, ALT, and $TNF-{\alpha}$ levels, increases that were ameliorated in the experimental groups. In addition, markedly increased expressions of cyclooxygenase (COX)-2 and its transcription factors, nuclear factor $(NF)-{\kappa}B$ and activator protein (AP)-1, were also significantly attenuated in the experimental groups. Compared to luteolin-7-O-glucoside, luteolin more potently ameliorated the levels of inflammatory mediators. Phase II enzymes levels and NF-E2 p45-related factor (Nrf)-2 activation that were decreased by GalN/LPS were increased by luteolin and luteolin-7-O-glucoside administration. In addition, compared to luteolin, luteolin-7-O-glucoside acted as a more potent inducer of changes in phase II enzymes. Liver histopathology results were consistent with the mediator and enzyme results. CONCLUSION: Luteolin and luteolin-7-O-glucoside protect against GalN/LPS-induced hepatotoxicity through the regulation of inflammatory mediators and phase II enzymes.

• Clinical and Experimental Reproductive Medicine
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• 제47권4호
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• pp.269-276
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• 2020

Objective: We investigated the impact of tyrosine kinase inhibitor (imatinib or dasatinib) coadministration with cyclophosphamide (Cp) on preantral follicle development in an in vitro mouse model. Methods: Seventy-three female BDF1 mice were allocated into four experimental groups: group A, saline; group B, Cp (25 mg/kg); group C, Cp (25 mg/kg) and imatinib (7.5 mg/kg); and group D, Cp (25 mg/kg) and dasatinib (7.5 mg/kg). Preantral follicles were isolated and cultured in vitro up to 12 days. Final oocyte acquisition and spindle integrity of metaphase II (MII) oocytes were assessed. Levels of 17β-estradiol and anti-Müllerian hormone (AMH) in the final spent media were measured by enzyme-linked immunosorbent assays, and the mRNA levels of Star, Sod1, Mapk3, and Casp3 in the final follicular cells were quantified by real-time polymerase chain reaction. Results: The percentage of MII oocytes per initiated follicle, the proportion of MII oocytes with normal spindles, and the 17β-estradiol level were similar in all four groups. The median AMH level in group B (7.74 ng/mL) was significantly lower than that in group A (10.84 ng/mL). However, the median AMH levels in group C (9.96 ng/mL) and group D (9.71 ng/mL) were similar to that in group A. The mRNA expression levels of Star, Sod1, Mapk3, and Casp3 were similar in all four groups. Conclusion: Coadministration of imatinib or dasatinib with Cp could preserve AMH production capacity in this in vitro mice preantral follicle culture model, and it did not affect MII oocyte acquisition.

• 생명과학회지
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• 제32권7호
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• pp.532-541
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• 2022

고지방식이(High fat diet, HFD)에 의해 유발되는 비만은 정상적인 마우스에서는 연구되지 않았지만 여러 유전자변형마우스의 전립선암(Prostate cancer)에 대한 강력한 위험인자 및 예후인자로 검증되었다. HFD-유도 비만이 정상적인 마우스에서 전립선암의 발생 및 진행에 영향을 미칠 수 있는지 여부를 조사하기 위해, 16주 동안 60% HFD 식이를 급여한 비만 C57BL/6N 마우스에서 전립선의 무게 및 조직학적 구조 변화와 암관련 단백질 발현을 분석하였다. 첫째, HFD 식이를 급여한 C57BL/6N 마우스는 체중, 장기의 무게, 지방축적, 혈청지질 수치의 증가 등을 포함한 비만증상을 성공적으로 유도되었다. 전립선의 무게는 No그룹에 비해 HFD-유도 비만마우스에서 유의미하게 증가하였다. 전립선의 4가지 엽들 중 외측전립선(Dorsolateral prostate, DLP)과 정낭(Seminal vesicle, SV)의 무게는 유의적인 변화가 없었지만, 복부전립선(Ventral prostate, VP)과 전방전립선(Anterior prostate, AP)의 무게는 No그룹보다 HFD-유도 비만마우스에서 증가하였다. 또한, 전립선의 조직학적 구조에서 과형성(Hyperplasia) 및 비호지킨림프종(Non-hodgkin's lymphoma, NHL)의 발생률은 HFD-유도 비만마우스에서 유의미하게 증가하였으며, 같은 그룹에서 AP의 상피두께도 증가하였다. HFD-유도 비만마우스에서 AKT (Protein kinase B) 신호경로에서 주요 단백질의 인산화수준이 유의미하게 증가했다. 따라서 이러한 결과는 HFD-유도 비만은 C57BL/6N 마우스에서 전립선암의 발생과 진행을 촉진할 수 있음을 시사한다.

• Journal of Ginseng Research
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• 제43권2호
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• pp.305-311
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• 2019

Background: Gintonin is a ginseng-derived exogenous ligand of the G protein-coupled lysophosphatidic acid (LPA) receptor. We previously reported that gintonin stimulates gliotransmitter release in primary cortical astrocytes. Astrocytes play key roles in the functions of neurovascular systems. Although vascular endothelial growth factor (VEGF) is known to influence the normal growth and maintenance of cranial blood vessels and the nervous system, there is little information about the effect of gintonin on VEGF regulation in primary astrocytes, under normal and hypoxic conditions. Methods: Using primary cortical astrocytes of mice, the effects of gintonin on the release, expression, and distribution of VEGF were examined. We further investigated whether the gintonin-mediated VEGF release protects astrocytes from hypoxia. Results: Gintonin administration stimulated the release and expression of VEGF from astrocytes in a concentration- and time-dependent manner. The gintonin-mediated increase in the release of VEGF was inhibited by the LPA1/3 receptor antagonist, Ki16425; phospholipase C inhibitor, U73122; inositol 1,4,5- triphosphate receptor antagonist, 2-APB; and intracellular $Ca^{2+}$ chelator, BAPTA. Hypoxia further stimulated astrocytic VEGF release. Gintonin treatment stimulated additional VEGF release and restored cell viability that had decreased due to hypoxia, via the VEGF receptor pathway. Altogether, the regulation of VEGF release and expression and astrocytic protection mediated by gintonin under hypoxia are achieved via the LPA receptor-VEGF signaling pathways. Conclusion: The present study shows that the gintonin-mediated regulation of VEGF in cortical astrocytes might be neuroprotective against hypoxic insults and could explain the molecular basis of the beneficial effects of ginseng on the central nervous system.

• 생명과학회지
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• 제24권12호
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• pp.1339-1344
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• 2014

염증성 장질환은 점막의 만성적 궤양과 염증을 동반하는 면역질환으로 알려져 있으며, 특히 TNF${\alpha}$와 같은 염증성 사이토카인은 주요한 생물학적 치료의 표적으로 이용되고 있다. 염증성 사이토카인의 유전자발현에서 전사물의 안정화는 매우 중요한 조절과정이며, 특히 본 연구에서는 이 안정화에 핵심적인 단백질인 HuR의 발현과 조직 분포에 대하여 동물모델과 환자의 조직에서 분석하였다. DSS를 처리함으로 유도되는 장염증 동물 모델에서 HuR 단백질의 발현량이 높았음을 확인했고, 점막의 상피조직 및 선조직 상피세포에서 상대적인 발현이 증대되었다. 또한 단백질의 활성측면에서 세포질로 이동된 HuR 단백질의 양도 상대적으로 증가하였다. 공간분포적으로 보면 DSS에 의한 화학적 점막자극에 의하여 초기에는 villi 하부에서의 발현정도가 상대적으로 villus 말단에 비하여 높게 유지되었다. 크론병 환자의 생검을 통하여 정상부위와 병변부위에서 HuR 단백질을 비교분석 하였다. 크론병 환자들의 병변에서는 지속적으로 HuR의 발현이 증대되어 있음을 확인했으며, 동물조직과 유사하게 병변부위의 장관상피세포 및 선 상피에서 주로 발현양이 높았다. 이러한 결과는 염증성 장질환에서의 HuR 단백질이 초기 염증성 인자의 발현에 중요한 역할이 예상되며, 구체적인 분자기전의 규명도 향후 기대된다. 이를 근간으로 하여 염증성 장질환의 진단과 치료의 표적개발에서 유용하게 응용하고자 한다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.51-51
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• 2001

Cocaine and amphetamine-regulated transcript (CART) is a satiety factor that is regulated by leptin. It was reported that the mice intracerebroventricularly injected with CART showed behavioral changes resembled with the typical behavioral alterations found in the mice carrying disorders in the brain serotonergic (5-HT) system. Hence, this study was conducted to find out the relationships between CART and 5-HT. We first examined the mRNA levels of CART after the injections of para-chlorophenylalanine (pCPA, 300 mg/kg i.p., single injection or daily for three consecutive days) in the rat brains by in situ hybridization using the mouse CART cDNA probe cloned in our laboratory. Systemic administrations of pCPA, a potent inhibitor of tryptophan hydroxylase, the rate limiting enzyme of 5-HT biosynthesis, acutely depletes the brain 5-HT transporter (5-HTT) in the dorsal raphe nucleus (DRN), which reuptakes terminal 5-HT. Results indicated that the mRNA level of CART significantly decreased in the arcuate nucleus, paraventricular nucleus, and lateral hypothalamic nucleus by three days of daily injection with pCPA with no noticeable change detected 24 hrs after the single injection. The message levels of 5-HTT in DRN decreased in both single and three days of injections. Secondly, to investigate whether CART affect to 5-HT, mouse genomic CART gene, which is consist of 3 exons and 2 introns and mouse neurofilament light (NF-L) chain promoter were cloned. Then, we constructed neuron specific expression vector, which was transfected into HeLa cell using lipid-mediated transfection system. Expression of GFP and CART linked to NF-L-chain promoter in the transfected HeLa cell were detected by using fluorescent microscope and RT-PCR. These results confirmed normal expression of DNA constructs in vitro. Then, to increase brain specific expression of CART in vivo transgenic mice carrying CART gene controlled the deleted NF-L-chain promoter were generated by the DNA microinjection into pronuclei of fertilized embryos. Transgenic mice were detected by Southern blot. Further study is necessary to examine CART expression and 5-HTT in these transgenic mice. Therefore, these results suggest that there maybe a positive molecular correlation between CART and 5-HT in responding to the stimuli.

• Asian Pacific Journal of Cancer Prevention
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• 제15권22호
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• pp.9805-9812
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• 2014

Radixin, a member of the ERM (ezrin-radixin-moesin) family, plays important roles in cell motility, invasion and tumor progression. It is expressed in a variety of normal and neoplastic cells, including many types of epithelial and lymphoid examples. However, its function in glioblastomas remains elusive. Thus, in this study, radixin gene expression was first examined in the glioblastoma cells, then suppressed with a lentivirus-mediated short-hairpin RNA (shRNA) method.We found that there were high levels of radixin expression in glioblastoma U251cells. Radixin shRNA caused down-regulation of radixin gene expression and when radixin-silenced cells were implanted into nude mice, tumor growth was significantly inhibited as compared to blank control cells or nonsense shRNA cells. In addition, microvessel density in the tumors was significantly reduced. Thrombospondin-1 (TSP-1) and E-cadherin were up-regulated in radixin- suppressed glioblastoma U251 cells. In contrast, MMP9 was down-regulated. Taken together, our findings suggest that radixin is involved in GBM cell migration and invasion, and implicate TSP-1, E-cadherin and MMP9 as metastasis-inducing factors.

• Nuclear Medicine and Molecular Imaging
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• 제41권1호
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• pp.42-48
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• 2007

목적: 이 연구에서는 폐 전이 종양을 영상화하기 위하여 흑색종의 폐 전이 종양 마우스 모델을 제작하고 영상 획득 전처리 조건을 개선하여 폐 전이 종양의 $[^{18}F]FDG$ 소동물 PET 영상을 획득하고자 하였으며, 임상 CT를 이용하여 전이 종양의 해부학적 위치를 확인하고자 하였다. 대상 및 방법: 정상 마우스의 $[^{18}F]$FDG 영상 획득 전 조건은 $16{\sim}22$시간 금식 하고 $30^{\circ}C$의 온도를 유지하며 $[^{18}F]FDG$ (7.4 MBq) 정맥 주사 후 서로 다른 마취제(Ketamine/Xylazine, Ke/Xy과 Isoflurane, Iso)로 $[^{18}F]FDG$ 섭취 60분 동안 유지한 후 20분간 $[^{18}F]FDG$ 소동물 PET 영상을 획득하였다. 혈중 포도당 농도를 보정한 포도당 표준 섭취 계수 영상을 이용하여 관심영역 대 배경비(lung to background ratio, L/B)를 구하여 평가하였다. C57BL/6 마우스에 B16-F10 세포를 정맥내 주사하여 제작한 폐전이 종양 마우스 모델은 정상 마우스의 영상 획득 조건과 동일한 조건에서 $[^{18}F]FDG$ 소동물 PET 영상을 획득하였으며, 임상 CT를 이용하여 획득된 해부학적 영상으로 폐 부위의 종양 위치를 확인하였다. 결과: 정상 마우스의 평균 혈중 포도당 농도는 Ke/Xy으로 마취한 군에서 $128.0{\pm}23.87\;mg/dL$이었으며 Iso으로 마취한 군에서는 $86.0{\pm}21.65\;mg/dL$로, Ke/Xy으로 마취한 군이 Iso로 마취한 군 보다 1.5 배 높은 혈중 포도당 농도를 나타내었다. 포도당 표준 섭취 계수 영상에서의 L/B는 Ke/Xy으로 마취한 군에서 $8.6{\pm}0.48$ 이었으며 Iso으로 마취한 군에서는 $12.1{\pm}0.63$로, Iso로 마취한 군이 Ke/Xy으로 마취한 군 보다 주변 정상조직과의 대조도가 높은 경향을 보였다. 폐 전이 종양 마우스에서는 Iso로 마취한 군이 Ke/Xy으로 마취한 군의 $[^{18}F]FDG$ 소동물 PET 영상보다 주변 조직의 $[^{18}F]FDG$ 섭취가 낮았다. 또한 해부학적 종양의 위치를 확인하기 위하여 임상 CT 영상과 융합한 결과 폐 전이 종양이 폐 부위에 위치함을 확인하였다. 결론: 마우스와 같은 소동물에서의 폐 부위 종양을 $[^{18}F]FDG$로 영상화하는데 있어서 금식, 온도유지, $[^{18}F]FDG$ 섭취 시간 동안의 마취제 조건 등을 고려하여야 하며, $[^{18}F]FDG$ 소동물 PET과 CT 영상의 융합은 폐 부위의 전이 종양을 확인하는데 유용할 것으로 기대된다.