• 한국식품영양과학회지
• /
• 제31권3호
• /
• pp.521-526
• /
• 2002

대두(백태,흑태)와 현미의 메탄올과 아세톤 추출물이 호르몬 의존형 유방암세포(MCF-7)와 호르몬 비의존형세포(MDA-MB-231)의 세포독성과 apoptosis에 미치는 영향을 살펴보았다 각 추출물별 25, 50, 100 ug/well의 농도로 24, 48, 72시간 배양 시 배양시간과 사용된 시료 모두 농도 의존적으로 유방암 세포생존율을 억제하는 것으로 나타났다. 특히 호르몬 의존형 세포인 MCF-7 에서는 현미의 아세톤 추출물이 낮은 농도에서 짧은 배양시간에도 그 효과가 나타났고 호르몬 비의존형 세포주 MDA-MB-231에서는 현미의 아세톤 및 메탄올 추출물의 효과가 다른 시료들에 비해 높게 나타났다. Apoptosis에 미치는 영향에서는 호르몬 비의존형 세포(MDA-MB-231)에서 메탄올추출물 처리군이 대조군에 비해 apoptosis된 세포가 유의적으로 증가한 것을 관찰할 수 있었다. 그러나 세포생존율 결과와는 다르게 호르몬의존형 세포와 호르몬비의존형 세포 모두에서 아세톤 처리군은 대조군에 비해 apptosis에 유의차를 나타내지 않았다. 이상의 결과에 의하면 이들 화합물이 소유한 암세포 성장억제 기전은 추출물내 함유된 화합물의 종류와 세포성장의 호르몬 의존도에 따라 다양한 것으로 사료된다.

• 한국수산과학회지
• /
• 제33권1호
• /
• pp.55-59
• /
• 2000

무지개 송어의 뇌하수체 및 배양액에 존재하는 GTH II 농도를 측정하기 위해 Avidin- Biotin complex를 이용한 sandwich EIA 계을 개발했다. Protein A sepharose affinity chromatography을 통해서 얻어진 연어 GTH II의 rabbit IgG에 biotinylation시킨 것 (Biotin-salmon GTH II rabbit IgG)을 제2 항체로 사용하였고, Non-Biotin salmon GTH II rabbit IgG는 단지 protein A sepharose affinity chromatography에서 얻어진 IgG를 제 1 항체로 사용하였다. EIA는 sandwich법에 의해서 이루어졌으며, 효소반응 기질로는 TMB(3,3'5,5-tetramethylbenzidine)를 이용했으며, 반응후 450 nm의 흡광도에서 automatic microplate reader로 측정하였다. 그 결과, $0.12\;{\~}\;125\;ng/ml$의 범위에서 용량반응곡선을 얻었으며, 측정감도 (최소 검출량)는 거의 0.58 ng/ml 정도 였다. 그리고 뇌하수체 추출물 및 배양액 각각의 희석곡선은 GTH II 표존곡선과 일치 하였다. 또한 이러한 GTH II의 표준곡선는 뇌하수체내 다른 peptide hormone와는 교차반응을 거의 나타내지 않았다. Testosterone을 처리한 미성숙 무지개 송어의 뇌하수체 세포배양계를 이용하여 sGnRH에 의한 GTH II 분비량을 본 sandwich EIA계와 RIA계를 비교 조사한 결과, 거의 같은 분비량을 나타냈을 뿐만아니라 같은 분비 pattern을 나타냈다. 이러한 결과로부터 본 sandwich법 EIA계에 의해서 연어과 어류의 뇌하수체 추출물 및 뇌하수체 배양액 중의 GTH II 함량 및 분비량을 측정하는데 있어서 안정된 assay계라고 생각되어진다.

• 대한한의학회지
• /
• 제34권2호
• /
• pp.29-40
• /
• 2013

Objectives: To investigate therapeutic mechanisms of Gyejibokryeong-hwan (GJBRH) against gynaecological diseases, articles on biological assay were gathered and analyzed. Methods: The articles were classified as being from domestic or international journals, and by their year of publication. The mechanisms of the biological effects against gynaecological diseases were noted. Results: Of the 14 articles analyzed, 13 were published in China and 1 was from Japan. GJBRH showed therapeutic effect against uterine and mammary gland diseases. Uterine-related diseases such as endometriosis, hysteromyoma, adenomyosis, cancer, and inflammation can be improved by the administration of GJBRH through anti-angiogenesis, anti-inflammation, the modulation of immune cell and immunoglobulin, and the regulation of hormone secretion. GJBRH also reduced mammary hyperplasia by regulating hormone and cytokine release. Conclusions: We speculate that the inhibitory effect against uterine and mammary gland diseases could be related to the therapeutic efficacy of GJBRH in improving gynaecological diseases.

• 한국식품조리과학회지
• /
• 제21권6호통권90호
• /
• pp.769-775
• /
• 2005

미성숙 흰쥐에서 누에 번데기 수용성과 지용성 추출 물 및 한약재가 첨가된 누에 번데기 수용성 추출물을 투여하여 에스트로젠 증식 관련 인자의 발현을 확인한 결과는 다음과 같다. 미성숙 흰쥐에서 누에 번데기 수용성 추출물(KW), 누에 번데기 지용성 추출물(KO) 및 KW에 하수오, 인삼, 울금을 7:1:1:1의 비율로 첨가한 복합체(MK)를 각각 100 mg/kg와 500 mg/kg의 농도로 30일간 경구투여한 결과 체중은 유의적인 변화를 나타내지 않았다. 체중에 대한 자궁의 무게는 정상 대조군에 대하여 실험군에서 KW500을 제외하고는 모두 유의적인 증가를 나타내었으며 K0500, KW100, MK100 순으로 각각 $0.49\%,\;0.48\%,\;0.44\%$의 높은 값을 나타내었다. 체중에 대한 난소의 무게는 정상 대조군에 대해 MK군을 제외하고 모두 유의적인 변화를 보였으며 KW100군이 가장 높은 값을 나타내었다. 미성숙 흰쥐의 혈청 내 AST와 ALT 활성 및 creatinine 농도는 모두 군 간에 유의적인 변화는 보이지 않았다. 즉 모든시료가 간과 신장에 독성을 나타내지 않은 결과이다. $ER\alpha$와 $ER\beta$의 발현을 densitometer로 수치화하여 정상대조군을 기준으로 발현의 증가와 억제를 백분율로 표시한 결과 KW100(100 mg/kg)와 MK500(500 mg/kg) 군이 $ER\alpha$의 발현을 유의적으로 증가시켰으며 KW100(100 mg/kg), KW500(500 mg/kg), 및 MK100(100 mg/kg)군이 $ER\beta$의 발현을 유의적으로 증가시켰다. 따라서 누에 번데기 수용성 추출물 및 한약재(인삼, 하수오, 울금)를 첨가한 누에 번데기 수용성 추출물이 생체에서 높은 에스트로젠 활성을 가지고 있는 것으로 사료된다.

• IMMUNE NETWORK
• /
• 제4권1호
• /
• pp.60-64
• /
• 2004

Background: Microglial cells, major immune effector cells in the central nervous system, become activated in neurodegenerative disorders. Activated microglial cells produce proinflammatory mediators such as nitric oxide (NO), tumor necrosis factor-$\alpha$ and interleukin-$1{\beta}$(IL-$1{\beta}$). These proinflammatory mediators have been shown to be significantly increased in the neurodegenerative disorders such as Alzhimer's disease and Pakinson's disease. It was known that one of the neurodegeneration source is stress and it is important to elucidate mechanisms of the stress response for understanding the stress-related disorders and developing improved treatments. Because one of the neuropeptide which plays a main role in regulating the stress response is corticotropin-releasing hormone (CRH), we analyzed the regulation of NO release by CRH in BV2 murine microglial cell as macrophage in the brain. Methods: First, we tested the CRH receptor expression in the mRNA levels by RT-PCR. To test the regulation of NO release by CRH, cells were treated with CRH and then NO release was measured by Griess reagent assay. Results: Our study demonstrated that CRH receptor 1 was expressed in BV2 murine microglial cells and CRH treatment enhanced NO production. Furthermore, additive effects of lipopolysaccaride (LPS) and CRH were confirmed in NO production time dependantly. Conclusion: Taken together, these data indicated that CRH is an important mediator to regulate NO release on microglial cells in the brain during stress.

• 대한간호학회지
• /
• 제29권2호
• /
• pp.445-455
• /
• 1999

This study has been conducted on the nonequivalent control group pretest-posttest design in quasi experimental basis and newly born premature infants from intensive care unit of G Medical University Hospital in Inchon Metropolitan were selected in two groups of 21 infants each. The first group for experimental and the other for control. Data has been collected form October 30, 1997 to August 29, 1998. For the experimental group tactile and kinesthetic stimulation was applied 2 times a day for 10 days(10 : 00~11 : 00 hours in the morning and 17 : 00~18 : 00 in the afternoon). As a weight weighing instrument. electronic indicator scale(Cas Co. korea) was used. To determine urine cortisol concentration level in stress hormone, radio immune assay method was used. And high performance liquid chlomatography was used to determine urine norepinephrine, concentration level To determine behavior status, tools developed by Anderson et at(1990) and remodeled by Kim Hee-Sook(1996) were used. Collected data were analyzed with the SAS program using x$^2$-test, student t-test, repeated measures ANOVA and paired t -test. The result were as follow. 1. As for the daily weight gain. the experimental group showed first change in weight and this group also showed higher weight in the average weight than the control group. Statistically, however, there was no significant factor between the two group. 2. The cortisol concentration in urine showed decrease in the experimental group norepinephrine concentration in urine showed increase in both experimental and control groups. No statistical significance was shown between the two groups. 3. In the aspect of behavior status. the experimental group showed statistical significance by showing inactive in the state of alert and conversion to a positive state than the control group. In conclusion, the sensory stimulation in this study showed a positive aspect through there was no statistical significance in the weight gain and urine stress hormone concentration. In the behavior status, there was statistical significance in the frequency of staying inactive in the state of alert and conversion to a positive state.

• 한국수정란이식학회지
• /
• 제28권2호
• /
• pp.149-155
• /
• 2013

Mullerian inhibiting substance (MIS) is a member of the TGF-${\beta}$ (transforming growth factor-${\beta}$) family whose members play key roles in development, suppression of tumour growth, and feedback control of the pituitary-gonadal hormone axis. MIS is expressed in a highly tissue-specific manner in which it is restricted to male Sertoli cells and female granulose cells. The serum levels of MIS in prenatal and postnatal ICR mice were measured using the enzyme-linked immuno-solvent assay (ELISA) using the MIS/AMH antibody. Mice were grouped by age: the significant periods were at the onset of development. During sex organ differentiation, no remarkable difference between female and male foetus MIS serum levels (both<0.1 ng/ml) was observed. However, MIS serum levels in pregnant mice markedly changed (4.5~12.2 ng/ml). After birth, postnatal female and male mice serum MIS levels changed considerably (male: <0.1~138.5 ng/ml, female: 5.3~103.4 ng/ml), and the changing phase were diametrically opposed (male: decreasing, female: fluctuating). These findings suggest that MIS may have strong associations with not only develop-ment but also puberty. For further studies, establishing the standard MIS serum levels is of importance. Our study provides the basic information for the study of MIS interactions with reproductive organ disability, cancer, and the effect of other hormone or menopause. We hypothesise that if MIS is regularly injected into middle-age women, meno-pause will be delayed. We detected that serum MIS concentration curves change with age. The changing phase is different between males and females, and this difference is significant after birth. Moreover, MIS mRNA is expressed during the developmental period (prenatal) and also in the postnatal period. This finding indicates that MIS may play a significant role in the developmental stage and in growth after birth.

• 한국발생생물학회지:발생과생식
• /
• 제25권4호
• /
• pp.225-234
• /
• 2021

The present study aimed to investigate the mechanism of cell surface receptor loss by two constitutively activating mutants (designated L469R, and D590Y) and two inactivating mutants (D417N and Y558F) of the luteinizing hormone receptor (LHR) in the Japanese eel Anguilla japonica, known to naturally occur in human LHR transmembrane domains. We investigated cell surface receptor loss using an enzyme-linked immunosorbent assay in HEK 293 cells. The expression level of wild-type eel LHR was considered to be 100%, and the expression levels of L469R and D417N were 97% and 101%, respectively, whereas the expression levels of D590Y and Y558F slightly increased to approximately 110% and 106%, respectively. The constitutively activating mutants L469R and D590Y exhibited a decrease in cell surface loss in a manner similar to that of wild-type eel LHR. The rates of loss of cell surface agonist-receptor complexes were observed to be very rapid (2.6-6.2 min) in both the wild-type eel LHR and activating mutants. However, cell surface receptor loss in the cells expressing inactivating mutants D417N and Y558F was slightly observed in the cells expressing inactivating mutants D417N and Y558F, despite treatment with a high concentration of agonist. These results provide important information on LHR function in fish and the regulation of mutations of highly conserved amino acids in glycoprotein hormone receptors.

• 대한약학회:학술대회논문집
• /
• 대한약학회 2000년도 춘계총회 및 학술대회
• /
• pp.127.1-127.1
• /
• 2000

• Toxicological Research
• /
• 제17권
• /
• pp.313-317
• /
• 2001

The Ministry of Health and Welfare of Japan has set up six research groups concerning the endocrine disrupting chemicals. One of these projects was "A study on development of testing methodology for health effects due to exposure of environmental endocrine disruptors". In this paper, three topics are described. In OECD collaboration for pre-validation of uterotrophic assay, the most sensitive response to ethnyl estradiol was noted in the ovarectomized rats treated subcutaneously for 7 days. Secondly, it was suggested that changes of the serum $\alpha_{2u}$-globulin level may be a sensitive parameter for detecting the estrogenic activities of chemicals. Finally, development of the sexually dimorphic nucleus of preoptic area in the brain oj male rats was inhibited by the treatment with estrogenic chemicals, and their masculine behaviors and reproductive abilities were impaired after sexual maturation. In conclusion, these parameters are considered to be sensitive endpoints for testing estrogenic chemicals.chemicals.