• The Korean Journal of Physiology and Pharmacology
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• 제2권6호
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• pp.771-778
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• 1998

To investigate interaction of angiotensin converting enzyme (ACE) inhibitor with local tissue renin- angiotensin system (RAS), changes in gene expression of the RAS components in various tissues in response to chronic administration of an ACE inhibitor, enalapril, were examined in Sprague-Dawley male rats. Enalapril was administered in their drinking water $(3{\sim}4\;mg/day)$ over 8 wk. Plasma and renal ACE activity increased significantly after 4 and 8 wk of enalapril treatment. Renin levels of the plasma and kidney of the enalapril-treated rats markedly increased after 4 wk and decreased thereafter, but still remained significantly higher than those of control rats. Kidney mRNA levels of renin markedly increased after 4 and 8 wk of enalapril treatment, but those of angiotensinogen and ANG II-receptor subtypes, $AT_{1A}$ and $AT_{1B}$, did not change significantly. The liver expressed genes for renin, angiotensinogen and $AT_{1A}$ receptor subtype, but $AT_{1B}$ receptor subtype mRNA was not detectable by RT-PCR. None of mRNA for these RAS components in the liver changed significantly by enalapril treatment. The hypothalamus showed mRNA expressions of renin, angiotensinogen, $AT_{1A}$ and $AT_{1B}$ receptor subtypes. $AT_{1A}$ receptor subtype mRNA was more abundant than $AT_{1B}$ receptor subtype in the hypothalamus as shown in the kidney. However, gene expression of the RAS components remained unchanged during 8-wk treatment of enalapril. In the present study, chronic ACE inhibition increased plasma and renal levels of ACE and renin, but did not affect mRNA levels of other RAS components such as angiotensinogen, ANG II receptor subtypes in the kidney. Gene levels of the RAS components in the liver and hypothalamus were not altered by chronic treatment of enalapril. These results suggest the differential expression of the RAS components in response to enalapril, and localized action and some degree of tissue specificity of enalapril.

• 동의생리병리학회지
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• 제20권2호
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• pp.383-387
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• 2006

To investigate the effect of GyeongshinhaeGihwan 1(GGT1) frequently used as an anti-obesity herbal medicine in oriental medicine on the expression of obesity-related genes, we measured the changes in mRNA levels of these genes by GGT1 in human growth hormone transgenic (hGHTg) obese female rats, and these effects by GGT1 were compared with those of reductil (RD), an anti-obesity drug approved by FDA. Rats received once daily oral administrations of autoclaved water, RD, or GGT1 for 8 weeks. At the end of study, rats were sacrificed and tissues were harvested. Total RNA from adipose tissue, liver and kidney was prepared and the mRNA levels for LPL (lipoprotein lipase), $PPAR{\gamma}$ (peroxisome proliferator activated receptor-gamma), $PPAR{\delta}$ (peroxisome proliferator activated receptor-delta), leptin, $TNF{\alpha}$ (tumor necrosis factor-alpha), and internal standard G3PDH (glyceraldehyde-3-phosphate dehydrogenase) were analyzed by RT-PCR. Compared with control group, $PPAR{\gamma}$ mRNA levels of liver and kidney were decreased in both RD and GGT1 groups, and the effects were more prominent in GGT1 group than in RD group, suggesting that GGT1 is effective in the inhibition of lipid storage by decreasing the $PPAR{\gamma}$ expression. $PPAR{\delta}$ mRNA levels of adipose tissue were increased by RD and GGT1 compared with DW, and the magnitude of increase were higher in GGT1 group than in RD group, indicating that GGT1 stimulates fatty acid oxidation and energy metabolism by activating $PPAR{\delta}$ expression. GGT1 group had higher concentrations of serum leptin, a well-known inhibitor of appetite, than control and RD groups. However, The mRNA levels of leptin, LPL, and $TNF{\alpha}$ were not changed by GGT1. These results indicate that GGT1 can prevent obesity in hGHTg obese female rats by down-regulating and up-regulating the mRNA expression of $PPAR{\gamma}$ and $PPAR{\delta}$, respectively, and that this anti-obesity effects were more pronounced in GGT1 group compared with RD group. In addition, GGT1 seems to inhibit obesity by increasing the circulating leptin levels.

• Animal cells and systems
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• 제5권3호
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• pp.217-224
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• 2001

The present study examines the expression and regulation of gonadotropin-releasing hormone (GnRH) and its receptor (GnRH-R) mRNA levels during mouse ovarian development. A fully processed, mature GnRH mRNA together with intron-containing primary transcripts was expressed in the immature mouse ovary as determined by Northern blot analysis and reverse transcription-polymerase chain reaction (RT-PCR). The size of ovarian GnRH mRNA was similar to that of hypothalamus, but its amount was much lower than that in the hypothalamus. Quantitative RT-PCR procedure also revealed the expression of GnRH-R mRNA in the ovary, but the estimated amount was a thousand-fold lower than that in the pituitary gland. We also examined the regulation of ovarian GnRH and GnRH-R mRNA levels during the follicular development induced by pregnant mare's serum gonadotropin (PMSG) and/or human chorionic gonadotropin (hCG). Ovarian luteinizing hormone receptor (LH-R) mRNA was abruptly increased st 48 h after the PMSG administration and rapidly decreased to the basal level thereafter. Ovarian GnRH mRNA level was slightly decreased at 48 h after the PMSG administration, and then returned to the basal value. GnRH-R mRNA level began to increase at 24 h after the PMSG treatment, decreased below the uninduced basal level at 48 h, and gradually increased thereafter. HCG administration did not alter ovarian GnRH mRNA level, while it blocked the PMSG-induced increase in GnRH mRNA level. Taken together, the present study demonstrates that the expression of GnRH and GnRH-R mRNA are regulated by gonadotropin during follicular development, suggesting possible intragonadal paracrine roles of GnRH and GnRH-R in the mouse ovarian development.

• 대한한의학회지
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• 제43권3호
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• pp.106-121
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• 2022

Objectives: The purpose of this study was to investigate the anti-inflammatory effect of Cornus Officinalis fruit extract(CE) and Cornus Officinalis Fruit Cheonghyeol Plus(CCP) in Human Umbilical Vein Endothelial Cell. Methods: We measured cell viability of CE, CCP and treated HUVEC with TNF-α. We measured the mRNA expression levels of KLF2, eNOS, MCP-1, ICAM-1, VCAM-1, the protein expression levels of KLF2, eNOS, MCP-1, ICAM-1, VCAM-1, and the protein phosphorylation level of ERK, JNK, p38 and the biomarker expression levels of MCP-1, ICAM-1, VCAM-1. Results: 1.CE incresed the mRNA, protein expression levels of KLF2, eNOS at concentrations of 100㎍/㎖ compared to the control group. CE decresed the mRNA, protein and biomarker expression levels of MCP-1,ICAM-1,VCAM-1 at concentrations of 100㎍/㎖ compared to the control group. CE decresed the protein phosphorylation level of p38 at concentrations of 100㎍/㎖ compared to the control group. 2. CCP incresed the mRNA, protein expression levels of KLF2, eNOS at concentrations of 100㎍/㎖ or more compared to the control group. CCP decresed the mRNA, protein and biomarker expression levels of MCP-1, ICAM-1, VCAM-1 at concentrations of 100㎍/㎖ or more compared to the control group. CCP decresed the protein phosphorylation level of ERK at concentrations of 100㎍/㎖ or more, p38 at concentrations of 200㎍/㎖ or more, and JNK at concentrations of 400㎍/㎖ compared to the control group. Conclusions: These results present that CE and CCP has anti-inflammatory effect in HUVEC. So, it could help treat or prevent inflammation in vein caused by dyslipidemia and contribute prevention of cardiovascular and cerebrovascular cerebrovascular diseases.

• The Korean Journal of Physiology and Pharmacology
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• 제7권2호
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• pp.79-84
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• 2003

We have previously reported that expression of the somatostatin receptor subtypes, sst1-5, is differentially regulated by growth hormone (GH)-releasing hormone (GHRH) and forskolin (FSK), in vitro. GHRH binds to membrane receptors selectively located on pituitary somatotropes, activates adenylyl cyclase (AC) and increases sst1 and sst2 and decreases sst5 mRNA levels, without significantly altering the expression of sst3 and sst4. In contrast FSK directly activates AC in all pituitary cell types and increases sst1 and sst2 mRNA levels and decreases sst3, sst4 and sst5 expression. Two explanations could account for these differential effects: 1) GHRH inhibits sst3 and sst4 expression in somatotropes, but this inhibitory effect is masked by expression of these receptors in unresponsive pituitary cell types, and 2) FSK inhibits sst3 and sst4 expression levels in pituitary cell types other than somatotropes. To differentiate between these two possibilities, somatotropes were sequentially labeled with monkey anti-rat GH antiserum, biotinylated goat anti-human IgG, and streptavidin-PE and subsequently purified by fluorescent-activated cell sorting (FACS). The resultant cell population consisted of 95% somatotropes, as determined by GH immunohistochemistry using a primary GH antiserum different from that used for FACS sorting. Purified somatotropes were cultured for 3 days and treated for 4 h with vehicle, GHRH (10 nM) or FSK ($10{\mu}M$). Total RNA was isolated by column extraction and specific receptor mRNA levels were determined by semi-quantitative multiplex RT-PCR. Under basal conditions, the relative expression levels of the various somatostatin receptor subtypes were sst2＞sst5＞sst3=sst1＞ sst4. GHRH treatment increased sst1 and sst2 mRNA levels and decreased sst3, sst4 and sst5 mRNA levels in purified somatotropes, comparable to the effects of FSK on purified somatotropes and mixed pituitary cell cultures. Taken together, these results demonstrate that GHRH acutely modulates the expression of all somatostatin receptor subtypes within GH-producing cells and its actions are likely mediated by activation of AC.

• 한국발생생물학회지:발생과생식
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• 제26권4호
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• pp.165-174
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• 2022

Three proteins [myosin heavy chain (MHC), filamin-C fragment (FIL-C), and actin 2 (ACT2)] were identified in adductor muscle from diploid and triploid Pacific oysters (Crassostrea gigas) and the relationship between the condition index (CI) and mRNA expression of these genes was investigated, together with the mRNA expression of molluscan insulin-related peptide (MIP), C. gigas insulin receptor-related receptor (CIR), and insulin-like growth factor binding protein complex acid labile subunit (IGFBP-ALS). Monthly changes in the CI were similar to the changes in the tissue weight rate in both groups. ACT2 and MHC mRNA expression was statistically higher in the triploid than the diploid, while FIL-C mRNA expression was significantly higher in the diploid (p<0.05). The MIP, CIR, and IGFBP-ALS mRNA expression of the diploid oysters were all significantly higher in July than in other months (p<0.05). The MIP, CIR, and IGFBP-ALS mRNA expression in the triploid oysters was high in July, but there were no significant differences (p>0.05). Changes in the expression levels of the genes investigated in this study could be used as intrinsic indicators of the annual growth, maturity, and spawning period of cultured diploid and triploid C. gigas in Tongyeong, Korea.

• 한국수정란이식학회지
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• 제29권3호
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• pp.297-303
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• 2014

The objective of this study was to determine the breed differences in the 3'-untranslated region (UTR) of MC1R mRNA, which may be used to distinguish Korean brindle cattle (Chikso) from other breeds. We investigated the relationship between the variation of 3'-UTR of the MC1R mRNA and coat color among different breeds and the Korean brindle cattle with different coat colors. MC1R mRNA expression levels were determined in accordance with the coat color and hair colors of the tail. Total cellular RNA was extracted from the hair follicles of the tails in Hanwoo, Korean brindle cattle, Holstein and $Hanwoo{\times}Holstein$ crossbred cattle. After cDNA synthesis, PCR was performed. Sequences of the 3'-UTR of MC1R mRNA were analyzed. The 3'-UTR of the MC1R mRNA from different breeds of cattle did not show any variations. There were no variations in the 3'-UTR of the MC1R mRNA in Korean brindle cattle with different coat colors. The levels of MC1R mRNA expression in hair follicles of the tail varied substantially among the Korean brindle cattle with different coat colors, except yellow coat color. Correlation between the MC1R mRNA expression in the hair follicles of the tail and coat color may be present in the Korean brindle cattle, but not between the variations of 3'-UTR of MC1R mRNA and coat color. Further studies to determine the regulation of MC1R mRNA expression from the hair follicles of different coat colors will be beneficial in clarifying the role of MC1R in the coat colors of the Korean brindle cattle.

• Tuberculosis and Respiratory Diseases
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• 제53권1호
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• pp.36-45
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• 2002

배 경 :포유동물은 고산소 노출되면 이에 적응하기 위해 성숙 폐장의 항산화 효소의 활성도가 증가하는 것으로 알려져 있다. 신생 폐장에서는 이들의 활성도가 뚜렷이 증가하고 이는 고산소 노출에 내성을 보이는 중요한 기전으로 알려져 있다. Peroxiredoxin은 세포내 항산화 효소로 세포내 많은 양이 존재하고 다양한 세포에 분포하고 있다. 그중 Prx I 및 II는 세포질에 존재하는 주된 동종 효소이다.본 연구는 고산소를 투여하여 성숙 백서의 폐장에서 Prx I과 Prx II mRNA 및 단백의 발현을 평가하여 신생 백서의 폐장에서의 발현과 비교하고자 하였다. 방 법 :성숙 백서와 임신 백서로부터 분만하여 얻은 신생 백서를 무작위로 고산소를 시간별로 투여후에 폐조직과 기관지폐포세척액을 얻었다. Prx I 및 Prx II mRNA는 Northernblot 방법으로 구하였으며 Actin mRNA을 내부 기준으로 하여 상대 발현을 평가하였다. Prx I 및 Prx II 단백은 Westernblot 방법으로 구하였으며 Actin 단백을 내부 기준으로 하여 상대 발현을 평가하였다. 결 과 : 성숙 백서에서 고산소 노출 후 24 시간에 Prx I mRNA 발현이 약간의 증가가 유도되었으며 신생 백서에서 고산소 노출 후 24 시간에 Prx I mRNA 발현이 현저한 증가가 유도되었다. 그러나 Prx II mRNA는 고산소 노출 내내 발현이 변화가 없었다. 성숙 및 신생 백서에서 고산소 노출 내내 Prx I 및 Prx II 단백의 발현이 변화가 없었으며 성숙 백서에서 고산소 노출 내내 기관지폐포세척액내 Prx I 및 Prx II 단백의 양의 변화가 없었다. 결 론 : 성숙 및 신생 백서에서 고산소 노출에 의해 Prx I 및 II의 발현이 전사 과정 에서 서로 다르게 조절된다. 성숙 백서보다 신생 백서에서 고산소 노출에 의한 Prx I mRNA의 뚜렷한 발현 증가는 신생 백서가 고산소에 내성을 보이는 하나의 기전으로 생각된다.

• 한국발생생물학회지:발생과생식
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• 제11권1호
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• pp.43-47
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• 2007

Transforming growth $factor-{\beta}(TGF-{\beta})$ 신호의 매개자 역할을 하는 Smad 계열 단백질은 발생과정에 중요한 역할을 한다고 알려져 있다. Estrogen receptor(ER)와 구조적으로 유사한 estrogen receptor-related receptor(ERR)은 포유동물에서 후기 배발생기에 외배엽 형성과 관련이 되어 있는 고아핵수용체이다. 본 연구에서는 해양무척추동물의 초기발생과정과 계절번식기 동안에 Smad3와 ERR의 유전자 발현이 발생과정과 성숙에 어떠한 연관성을 갖고 있는지 알아보기 위하여, 동해안 연안에 주로 서식하는 극피동물문 둥근성게과 둥근성게(Strongylocentrotus nudus)를 재료로 하여 계절별 및 배발생 과정중에 Smad3와 $ERR{\beta}$ like 1의 mRNA 농도를 real-time PCR 방법으로 조사하였다. Smad3 mRNA는 샘플링을 시작한 2004년 2월의 생식소와 비교하면 4월부터 그 농도가 증가하기 시작하여 6월까지 증가하였으며, 산란기인 8월에 감소하였다가 10월부터 12월까지 높은 수준을 유지하였다. $ERR{\beta}$ like 1 mRNA는 6월까지 낮은 수준이었으나, 산란기인 8월에 급증한 후 다시 감소하였다. 수정란부터 초기 유생기까지 발생과정을 분석한 결과, Smad3 mRNA는 8세포기 및 16세포기에 높은 발현이 관측되었다. 한편, $ERR{\beta}$ like 1 mRNA는 포배기, 낭배기, 초기 유생기에 현저하게 높은 발현패턴을 보였다. 이상의 결과로부터 둥근성게의 산란기 및 발생배의 발생후기에 $ERR{\beta}$ like 1이 중요한 역할을 담당할 것으로 추정되며, 초기 난할시기에는 Smad3의 관련성이 시사되었다.

• Toxicological Research
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• 제33권2호
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• pp.149-156
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• 2017

The purpose of this study was to investigate the wound healing effect of acai berry water extracts (ABWE) and a possible underlying mechanism involved in its action using various in vitro and in vivo models. The wound healing effect of ABWE was evaluated by migration assay using HS68 fibroblast cells. In addition, its effect on mRNA expression of procollagen, fibronectin, and MMP-1 was determined. Moreover, the wound healing effect of ABWE was evaluated in in vivo wound models through macroscopic and microscopic observation. In addition, mRNA expression levels of wound related genes were determined. Results revealed that ABWE was not cytotoxic. It increased migration of HS68 fibroblast cells. ABWE increased mRNA expression levels of fibronectin but decreased the mRNA expression levels of MMP-1. ABWE also showed significantly potent wound healing effect in vivo based on macroscopic and histopathological observation and mRNA expression evaluation for wound related genes. Taken together, our results indicated that ABWE might have potential as a wound healing agent.