• 대한약리학회지
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• 제32권2호
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• pp.233-241
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• 1996

Previous studies have shown that glucocorticoid suppresses microsomal epoxide hydrolase(EH) gene expression and that EH expression is altered during pregnancy. The effects of progesterone on the expression of rat EH and certain glutathione S-transferase(GST) genes were examined in this study. Northern RNA blot analysis revealed that progesterone was effective in increasing hepatic EH mRNA levels at 12 h to 48 h after treatment with a maximal 9-fold increase being noted at 12 h time point. Nonetheless, multiple daily treatment with progesterone rather caused minimal relative increases in EH mRNA levels. GST Ya and Yb1/2 mRNA levels were also transiently elevated at 12 h after progesterone treatment, followed by gradual decreases from the maximal Increases at day 1, 2 and 5 post-treatment. These changes in EH and GST mRNA levels were noted only at a relatively high dose of progesterone. Furthermore, immunoblot analyses showed that rats treated with progesterone for 5 days failed to show EH or GST induction, indicating that progesterone-induced alterations in EH and GST mRNA levels do not reflect bona fide induction of the detoxifying enzymes. Concomitant progesterone treatment of rats with the known EH inducers including ketoconazole and clotrimazole failed to additively nor antagonistically alter EH mRNA levels. In contrast, dexamethasone substantially reduced ketoconazole- or clotrimazole-inducible EH expression. These results showed that progesterone stimulates the EH, GST Ya and Yb1/2 gene expression at early times followed by marked reduction in the RNA levels from the maximum after multiple treatment and that the changes in mRNA do not necessarily reflect induction of the proteins.

• 한국환경과학회지
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• 제29권6호
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• pp.633-641
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• 2020

Water temperature is one of the most important factors of fish survival, affecting the habitat, migration route, development, and reproduction. This experiment studied the induction level of heat shock protein (HSP70) mRNA and protein in a walleye pollock (Gadus chalcogrammus) primary hepatocyte culture based on different temperatures. Hepatocytes were attached at 7.5℃ for 24 hours. Hsp70 induction levels were then measured for 48 hours at 5, 8, 11, 14, and 17℃. The induction level was lowest at 5℃ and generally increased with temperature until 14℃. The induction level was reduced at 17℃, indicating that 14℃ is the highest tolerable temperature for hepatocytes. These data indicate that primary hepatocyte cell culture is under no stress at 5 and 8℃. Temperatures greater than 11℃ induce stress, showing similar induction patterns in both mRNA and protein in hepatocytes. The results suggest that 14℃ is the maximum internal defense temperature of walleye pollock survival.

• Toxicological Research
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• 제17권3호
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• pp.181-186
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• 2001

The estrogenic potency of bisphenol A using reverse trancriptase-PCR response of liver vitellogenin mRNA in male carp was studied. For this, six combination of primers which were synthesized on the basis of cDNA consensus region of various species, were evaluated and one pair of primers was selected as the best to show 286 bp size-transcript. By using the selected primers, vitellogenin mRNA induction in carp treated with bisphenol A was measured and the chemical showed dose-and time-dependent Induction response. From this result, it was concluded that RT-PCR technique wing the selected primers in this study can be wed to monitor the estrogenic effects exerted In carp living in Korean freshwater.

• Environmental Analysis Health and Toxicology
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• 제16권1호
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• pp.29-34
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• 2001

The estrogenic potency of di -2-ethylhexyl phthalate (DEHP) using reverse transcriptase-PCR respouse of liver vitellogenin mRNA in male African clawed frog (Xenopus laevis) was studied. Male frogs were injected with DEHP at dose of 300$\mu\textrm{g}$/kg and 300 mg/kg body weight through the dorsal lymph sac. After 4 days, using suitable pair of RT-PCR primers, vitellogenin mRNA induction in the liver was measured and DEHP showed vitellogenin mRNA induction in only the group treated with 300 mg/kg. Any significant histological abnormalities by the exposure of DEHP was not shown in both testis and liver.

• 한국환경과학회:학술대회논문집
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• 한국환경과학회 2003년도 International Symposium on Clean Environment
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• pp.159-164
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• 2003

Effects of Al on vitellogenin (VTG) and VTG mRNA induction by estradiol-17 $\beta$($E_2$) were examined in primary hepatocyte culture of rainbow trout. Hepatocytes were precultured for 2 days and then E2 ($2{\times}10^{-6}$M) and Al ($10^{-6}-10^{-4}$M) were simultaneously added to the incubation medium. Hepatocytes were cultured for 5 more days. Media and hepatocytes were then analyzed by SDS-PAGE and Northern blotting for VTG and VTG mRNA, respectively. These metal had no appreciable effect on the viability of hepatocytes in culture. However, Al interfered with VTG production and VTG mRNA expression. Al reduced VTG production in a concentration-dependent way and a significant reduction accurred at Al concentrations greater than $5{\times}10^{-5}$M. VTG mRNA expression also decreased with a negative correlation with Al concentration (r＝-0.98). These results suggest that Al inhibit VTG production at the transcriptional level to reduce VTG mRNA expression.

• Biomolecules & Therapeutics
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• 제26권5호
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• pp.446-457
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• 2018

Adiponectin, a hormone predominantly originated from adipose tissue, has exhibited potent anti-inflammatory properties. Accumulating evidence suggests that autophagy induction plays a crucial role in anti-inflammatory responses by adiponectin. However, underlying molecular mechanisms are still largely unknown. Association of Bcl-2 with Beclin-1, an autophagy activating protein, prevents autophagy induction. We have previously shown that adiponectin-induced autophagy activation is mediated through inhibition of interaction between Bcl-2 and Beclin-1. In the present study, we examined the molecular mechanisms by which adiponectin modulates association of Bcl-2 and Beclin-1 in macrophages. Herein, we demonstrated that globular adiponectin (gAcrp) induced increase in the expression of AUF1 and ZFP36L1, which act as mRNA destabilizing proteins, both in RAW 264.7 macrophages and primary peritoneal macrophages. In addition, gene silencing of AUF1 and ZFP36L1 caused restoration of decrease in Bcl-2 expression and Bcl-2 mRNA half-life by gAcrp, indicating crucial roles of AUF1 and ZFP36L1 induction in Bcl-2 mRNA destabilization by gAcrp. Moreover, knock-down of AUF1 and ZFP36L1 enhanced interaction of Bcl-2 with Beclin-1, and subsequently prevented gAcrp-induced autophagy activation, suggesting that AUF1 and ZFP36L1 induction mediates gAcrp-induced autophagy activation via Bcl-2 mRNA destabilization. Furthermore, suppressive effects of gAcrp on LPS-stimulated inflammatory mediators expression were prevented by gene silencing of AUF1 and ZFP36L1 in macrophages. Taken together, these results suggest that AUF1 and ZFP36L1 induction critically contributes to autophagy induction by gAcrp and are promising targets for anti-inflammatory responses by gAcrp.

• Environmental Analysis Health and Toxicology
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• 제18권1호
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• pp.15-19
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• 2003

Metallothionein gene expression activity of cadmium was investigated in a human lung epithelial cell line. Cells, grown to near confluence, were exposed to 0∼10 ${\mu}$M Cd metal for 6 hours. Cadmium did not cause morphological alteration in lung epithelial cells that are characteristic of cell damages such as cell shrinkage, detachment of the cell from its neighbors, cytoplasmic and chromatic condensation. However, metallothionein genes of MT-1 and MT-2 were rapidly induced in the treated cell measured by RT-PCR. Regarding the induction pattern of motallothionein mRNA, MT-1 mRNA was induced in a dependent manner. MT-2 mRNA induction, which was measured using oligo primers based on cDNA of human reticulocytes, seemed to be slightly increased in low doses but decreased at high concentration used in the experiment.

• Fisheries and Aquatic Sciences
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• 제4권4호
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• pp.186-191
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• 2001

Vitellogenin (VTG) and VTG mRNA induction by $estradiol-17\beta\;(E_2)$ were examined in the primary cultures of hepatocyte in the rainbow trout. Hepatocytes were precultured for 2 days, then $E_2$ was added and cultured for another 5 days. Media and hepatocytes were then analyzed by electrophoresis and Northern blotting for VTG and VTG mRNA, respectively. The hepatocytes were formed a few aggregates within 5 days without further spreading to a monolayer. Cell viability and high DNA content were maintained during the incubation. The hepatocyte culture with E2 induced a weak VTG band at a molecular weight of 175kDa on Day 2 after $E_2$ addition. The relative amount of VTG was expressed in percentage of total protein concentrations. VTG was gradually increased as $1.9\%$ on Day 2, $6.3\%$ on Day 4 and $7.3\%$ on Day 5. VTG mRNA band was detected at about 6.6 kb in the culture with $E_2$ at day 1 of culture. The level of VTG mRNA expression linearly increased with time until Day 5 (r=0.97).

• 대한의생명과학회지
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• 제10권4호
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• pp.479-483
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• 2004

The aim of this study was carried to the induction of follistatin-like 1 (FSTL1) in rat myometrium tissue by exogenous estrogen. PCR was performed by using estrogen treatment after ovariectomy of myometrium mRNA and only ovariectomy of myometrium mRNA and on ovariectomy of myometrium mRNA in rat normal uterus tissue. The PCR products were visualized on agarose gel (1.2％) electrophoresis. FSTL1 mRNA expression level was significantly higher (**P<0.001 or *P<0.05) in estrogen treatment after ovariectomy than in only ovariectomy, and was significantly higher (**P<0.001) in no ovariectomy than in estrogen with treatment after ovariectomy. In conclusion, Although the mechanisms of FSTL1 gene were uncertain, It also might be inducted by exogenous estrogen. And also it is thought to be related to the estorgen mechanism.

• 한국환경농학회지
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• 제36권4호
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• pp.241-248
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• 2017

본 연구는 최근 RNAi 기반 LMO의 연구 개발이 활발히 진행됨에 따라 향후 이러한 기술을 이용한 LMO의 유해성 및 자연생태계 위해성평가가 필요할 때 실험실 수준에서 dsRNA를 대량으로 발현시키는 시스템을 확립하고, 수분(화분)매개 곤충인 꿀벌을 대상으로 유해성평가 시험을 수행하는 방법을 제시하고자 하였다. L4440 vector에 Snf7과 GFP 유전자를 클로닝한 plasmid를 HT115 (DE3) 대장균에 형질 전환한 후 온도, 배양시간, IPTG 농도를 각기 다르게 하여 최적의 발현조건을 탐색한 결과 $37^{\circ}C$, 0.4 mM IPTG, 4시간의 배양시간에서 가장 많은 양의 dsRNA가 발현됨을 확인하였다. 국내 외 제시된 꿀벌 위해성평가 가이드라인을 바탕으로 대장균에서 분리한 dsRNA를 꿀벌 성충에 급성섭식으로 처리한 결과 생사율과 일반중독증상에서 차이를 보이지 않는 것으로 보아 대장균으로부터 분리한 Snf7 dsRNA와 GFP dsRNA는 꿀벌 성충에 유해하지 않음을 알 수 있었다. 본 연구를 통해 dsRNA 물질의 유해성평가 및 자연생태계 위해성 평가를 위한 대량 추출 방법과 위해성평가 대상종의 사육 및 물질 처리 방법을 확립하여 향후 이뤄질 dsRNA의 꿀벌 위해성평가에 활용될 것으로 사료된다.