• Toxicological Research
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• 제8권2호
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• pp.343-357
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• 1992

Tumor necrosis factor-${\alpha}$(TNF), a polypeptide hormone secreted primarily by activated macrophages, was originally identified on the basis of its ability to cause hemorrhagic necrosis and tumor regression in vivo. Subsequently, TNF has been shown to be an important component of the host responses to infection and cancer and may mediate the wasting syndrome known as cachexia. These systemic actions of TNF are reflected in its diverse effects on target cells in vitro. TNF initiates its diverse cellular actions by binding to specific cell surface receptors. Although TNF receptors have been identified on most of animal cells, regulation of these receptors and the mechanisms which transduce TNF receptor binding into cellular responses are not well understood. Therefore, in the present study, the mechanisms how TNF receptors are being regulated and how TNF receptor binding is being transduced into cellular responses were investigated in rat liver plasma membranes (PM) and ME-180 human cervical carcinoma cell lines. $^{125}I$-TNF bound to high ($K_d=1.51{\pm}0.35nM$)affinity receptors in rat liver PM. Solubilization of PM with 1% Triton X-100 increased both high affinity (from $0.33{\pm}0.04\;to\;1.67{\pm}0.05$ pmoles/mg protein) and low affinity (from $1.92{\pm}0.16\;to\;7.57{\pm}0.50$ pmoles/mg protein) TNF binding without affecting the affinities for TNF, suggesting the presence of a large latent pool of TNF receptors. Affinity labeling of receptors whether from PM or solubilized PM resulted in cross-linking of $^{125}I$-TNF into $M_r$ 130 kDa, 90 kDa and 66kDa complexes. Thus, the properties of the latent TNF receptors were similar to those initially accessible to TNF. To determine if exposure of latent receptors is regulated by TNF, $^{125}I$-TNF binding to control and TNF-pretreated membranes were assayed. Specific binding was increased by pretreatment with TNF (P<0.05), demonstrating that hepatic PM contains latent TNF receptors whose exposure is promoted by TNF. Homologous up-regulation of TNF receptors may, in part, be responsible for sustained hepatic responsiveness during chronic exposure to TNF. As a next step, the post-receptor events induced by TNF were examined. Although the signal transduction pathways for TNF have not been delineated clearly, the actions of many other hormones are mediated by the reversible phosphorylation of specific enzymes or target proteins. The present study demonstrated that TNF induces phosphorylation of 28 kDa protein (p28). Two dimensional soidum dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) resolved the 28kDa phosphoprotein into two isoforms having pIs of 6.2 and 6.1. The pIs and relative molecular weight of p28 were consistent with those of a previously characterized mRNA cap binding protein. mRNA cap binding proteins are a class of translation initiation factors that recognize the 7-methylguanosine cap structure found on the 5' end of eukaryotic mRNAs. In vitro, these proteins are defined by their specific elution from affinity columns composed of 7-methylguanosine 5'-triphosphate($m^7$GTP)-Sepharose. Affinity purification of mRNA cap binding proteins from control and TNF treated ME-180 cells proved that TNF rapidly stimulates phosphorylation of an mRNA cap binding protein. Phosphorylation occurred in several cell types that are important in vitro models of TNF action. The mRNA cap binding protein phosphorylated in response to TNF treatment was purifice, sequenced, and identified as the proto-oncogene product eukaryotic initiation factor-4E(eIF-4E). These data show that phosphorylation of a key component of the cellular translational machinery is a common early event in the diverse cellular actions of TNF.

• 대한융합한의학회지
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• 제5권1호
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• pp.55-60
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• 2023

Objectives: This study was performed to investigate the effect of TJGB on the liver of high-fat diet (HFD)-fed mice and the cell viability of HepG2 cells. Methods: After a week adaptation, 8-week-old C57BL/6N mice were fed with a 45% HFD or normal diet for 3 weeks. For the next 9 weeks, the mice were divided into 6 groups: normal diet group; HFD group; HFD plus orlistat group; HFD plus Ephedra sinica Stapf (ES) group; HFD plus low dose of TJGB group; HFD plus high dose of TJGB group. To estimate the effect of TJGB in the liver of HFD-fed mice, the protein expressions of phospho-acetyl-CoA carboxylase (p-ACC) and liver X Receptor (LXR) were determined by Western blot assay. The cell viability of ES and TJG was also evaluated in HepG2 cells. Results: The administration of TJGB had little effect on the protein expressions of p-ACC and LXR in the liver of HFD-fed mice. And the cytotoxicity was showed above 7.8 ㎍/mL in HepG2 cells. Conclusion: Further research is needed to evaluate the mechanism of TJGB on hepatic steatosis and cytotoxicity in HepG2 cells.

• IMMUNE NETWORK
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• 제8권1호
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• pp.1-6
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• 2008

Apoptosis signal-regulating kinase 1 (ASK1), a mitogen- activated protein kinase kinase kinase, plays pivotal roles in stress responses. In addition, ASK1 has emerged as a key regulator of immune responses elicited by pathogen-associated molecular patterns (PAMPs) and endogenous danger signals. Recent studies have demonstrated that reactive oxygen species (ROS)-dependent activation of ASK1 is required for LPS-stimulated cytokine production as well as extracellular ATP-induced apoptosis in immune cells. The mechanism of ROS-dependent regulation of ASK1 activity by thioredoxin and TRAFs has been well characterized. In this review, we focus on the molecular details of the activation of ASK1 and its involvement in innate immunity.

• Childhood Kidney Diseases
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• 제7권1호
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• pp.52-59
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• 2003

목적 : 레닌-안지오텐신계는 신장의 성장과 발달에 중요한 역할을 하는 것으로 알려져 있으며, 안지오텐신 II는 신장형성기 동안 중요한 역할을 담당한다. 저자들은 터너 증후군 환자에서 신기형의 발생빈도를 조사하고, 신기형이 동반된 군과 동반되지 않은 군으로 분류하여 양군 사이에 레닌-안지오텐신계 유전자의 분포에 차이가 있는지를 조사하였다. 방법 : 말초혈액의 임파구를 사용하여 세포분 석학적 방법으로 진단된 33명의 터너 증후군 환자를 대상으로 신요로계의 기형여부를 초음파검사와 경정맥 요로 조영 검사, DMSA scan으로 진단하고, 더불어 안지오텐신전환효소의 유전자형, 안지오텐시노겐 유전자형, 안지오텐신 수용체유전자형의 분포를 핵산증폭법을 사용해 조사하였다. 결과 : 33명의 터너 증후군 환자 중 12명에서 신기형이 동반되어 전체적으로 36.4%의 빈도를 나타내었다. 핵형에 따른 신기형의 빈도를 살펴보면 전형적인 45,X형의 경우는 18명 중 8명에서 관찰되어 44.4%의 빈도를 나타내었고, 모자이시즘과 X 염색체구조상의 이상인 경우에는 15명 중 4명에서 관찰되어 26.7%의 빈도를 나타내어, 전형적인 45,X형의 경우에서 발생빈도가 높았으나 통계적으로 유의하지는 않았다(P>0.05). ACE 유전자형의 분포는 신기형 동반군에서 DD형이 0%, ID형이 73%, 그리고 II형이 27%이었으며, 신기형이 동반되지 않은 군에서는 DD형이 11%, ID형이 56%, II형이 33%로 양군 사이에는 ACE 유전자형의 분포는 유의한 차이가 없었다. AGT M235T 유전자형의 분포는 신기형 동반군에서 MM형이 82%, MT형이 8%, 그리고 TT형이 0%이었으며, 신기형이 동반되지 않은 군에서는 MM형이 56%, MT형이 33%, TT형이 11%로 양군 사이에는 AGT M235T 유전자형의 분포는 유의한 차이가 없었다. ATR 1 유전자형의 분포는 신기형 동반군에서 AA형이 91%, AC형이 9%, 그리고 CC형이 0%이었으며, 신기형이 동반되지 않은 군에서는 AA형이 94%, AC형이 6%, CC형이 0%로 양군 사이에는 ATR 1 유전자형의 분포도 유의한 차이가 없었다. 결론 : 터너증후군 환자에서 신기형의 동반율은 36.4%였으며, 45,X형에서 발생빈도가 높았으나 통계적으로 유의하지는 않았다. 신기형이 동반된 군과 동반되지 않은 군 사이에는 레닌-안지오텐신계 유전자의 분포는 모두 유의한 차이가 관찰되지 알았다.

• Asian-Australasian Journal of Animal Sciences
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• 제27권12호
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• pp.1695-1704
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• 2014

Maternal malnutrition during pregnancy may give rise to female offspring with disrupted ovary functions in adult age. Neonatal ovary development predisposes adult ovary function, yet the effect of maternal nutrition on the neonatal ovary has not been described. Therefore, here we show the impact of maternal protein restriction on the expression of folliculogenic and steroidogenic genes, their regulatory microRNAs and promoter DNA methylation in the ovary of neonatal piglets. Sows were fed either standard-protein (SP, 15% crude protein) or low-protein (LP, 7.5% crude protein) diets throughout gestation. Female piglets born to LP sows showed significantly decreased ovary weight relative to body weight (p<0.05) at birth, which was accompanied with an increased serum estradiol level (p<0.05). The LP piglets demonstrated higher ratio of bcl-2 associated X protein/B cell lymphoma/leukemia-2 mRNA (p<0.01), which was associated with up-regulated mRNA expression of bone morphogenic protein 4 (BMP4) (p<0.05) and proliferating cell nuclear antigen (PCNA) (p<0.05). The steroidogenic gene, cytochrome P450 aromatase (CYP19A1) was significantly down-regulated (p<0.05) in LP piglets. The alterations in ovarian gene expression were associated with a significant down-regulation of follicle-stimulating hormone receptor mRNA expression (p<0.05) in LP piglets. Moreover, three microRNAs, including miR-423-5p targeting both CYP19A1 and PCNA, miR-378 targeting CYP19A1 and miR-210 targeting BMP4, were significantly down-regulated (p<0.05) in the ovary of LP piglets. These results suggest that microRNAs are involved in mediating the effect of maternal protein restriction on ovarian function through regulating the expression of folliculogenic and steroidogenic genes in newborn piglets.

• Asian-Australasian Journal of Animal Sciences
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• 제17권5호
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• pp.598-602
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• 2004

The Tongcheng pig breed is a famous Chinese indigenous breed. The Ministry of Agriculture of China has filed it as 1 of 19 national key conservation breeds selected from more than 100 Chinese indigenous pig breeds in 2000. In order to improve the reproductive performance, it has been intensively selected to increase the litter size for about 10 years. The population randomly sampled from conservation nucleus of eight families in the Tongcheng pigs was genotyped for identification of their estrogen receptor locus polymorphisms with the PCR-RFLPs method. Only AB heterozygotes and BB homozygotes were detected, and $X^2$ test demonstrated that the locus was in disequilibrium at a significant level (p<0.05). In the present paper, the litter sizes in different parities were regarded as different traits. Holistic status of other unspecific and unidentified genes was estimated by using the statistical methods. Coefficients of kurtosis and skewness showed that the litter size still presented segregating characteristic in the 2nd, 5th, 7th, 8th and 9th parities. Analysis of homogeneity of variance between families confirmed the results for the 5th, 7th and 8th parities. The heritability of litter size for the 1st to 10th parities was estimated with paternal half-sib model and individual estimated breeding values (EBVs) were evaluated by a single trait animal model as well. We found that the averages of EBVs for litter size in each parity did not differ significantly between genotypes, despite the significant difference for original phenotype records in the 3rd, 4th and 5th parities (p<0.05 or p<0.01). The results may be explained by the deduction that the polymorphisms of ESR locus are no longer the important genetic base of litter size variation when the frequency of allele B accumulated in the experience of selection procedure, and further conferring that there exist special genes associated with litter size in the recent Tongcheng pigs population can be made.

• 대한방사선기술학회지:방사선기술과학
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• 제33권4호
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• pp.327-334
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• 2010

전리방사선의 피폭에 의한 암 유발확률은 소아가 성인에 비해 크기 때문에 소아 X선 검사 시의 환자선량을 정확히 파악하는 일은 중요하다. 그러나 우리나라는 소아 환자선량에 대한 연구가 활발하지 않다. 그러므로 본 연구에서는 7세 미만 소아를 대상으로 전리함을 이용하여 흉부검사 시의 입사피부선량(entrance skin dose, ESD)을 측정하여 나이, 키, 체중, 가슴두께와 ESD의 관계를 비교한 결과는 다음과 같았다. 소아는 나이에 따라서 키, 체중 및 가슴두께가 비슷한 양상으로 증가하므로, 흉부촬영 시 측정하기 힘든 가슴두께 대신에 나이에 따라서 촬영조건을 설정해도 무방함을 알 수 있었다. 2세 미만 소아의 흉부 A-P 검사 시, kVp는 A병원에서 더 높았으나, mAs는 반대로 B병원에서 높아, ESD 값이 B병원에서 약 1.7배 높았다. 그러나 4세 이상 소아의 흉부 P-A 검사 시에는 mAs는 같았고 kVp는 B 병원에서 7 kVp 높았으나, ESD값은 FID가 먼 B병원(180 cm)에서보다 더 가까운 A병원(130 cm)에서 1.4배 정도 높았다. 또한 같은 나이라도 A-P가 P-A보다 ESD값이 높았다. 나이별에 따른 ESD값을 보면, 1세 미만은 $154{\mu}Gy$, 1세에서 4세 미만은 $194{\mu}Gy$, 4세에서 7세 미만은 $138{\mu}Gy$으로 나타났다. 이 값은 일본의 JART의 권고량($200{\mu}Gy$)보다 낮으나, EC(유럽위원회)나 영국의 NRPB의 권고량보다 높으며, 2009년 12월에 식품의약품안전평가원에 제출된 용역보고서의 진단참고준위(5세 소아에서의 중간값이 $100{\mu}Gy$)보다 높다. 결론적으로 ESD는 X선장치 시스템의 차이보다는 방사선사의 경험적 실행에 의한 촬영조건의 차이에 의해서 크게 달라짐을 알 수 있었으며, 또한 나이가 많다고 더 많은 선량을 받는 것은 아니다. 따라서 나이에 따른 소아의 적정 참고준위의 확립과 점진적인 환자선량의 저감화가 반드시 필요하다.

• The Korean Journal of Physiology and Pharmacology
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• 제26권6호
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• pp.447-456
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• 2022

The present study was carried out to investigate the effect of Arctigenin on cell growth and the mechanism of cell death elicited by Arctigenin were examined in FaDu human pharyngeal carcinoma cells. To determine the apoptotic activity of Arctigenin in FaDu human pharyngeal carcinoma cells, cell viability assay, DAPI staining, caspase activation analysis, and immunoblotting were performed. Arctigenin inhibited the growth of cells in a dose-dependent manner and induced nuclear condensation and fragmentation. Arctigenin-treated cells showed caspase-3/7 activation and increased apoptosis versus control cells. FasL, a death ligand associated with extrinsic apoptotic signaling pathways, was up-regulated by Arctigenin treatment. Moreover, caspase-8, a part of the extrinsic apoptotic pathway, was activated by Arctigenin treatments. Expressions of anti-apoptotic factors such as Bcl-2 and Bcl-xL, components of the mitochondria-dependent intrinsic apoptosis pathway, significantly decreased following Arctigenin treatment. The expressions of pro-apoptotic factors such as BAX, BAD and caspase-9, and tumor suppressor -53 increased by Arctigenin treatments. In addition, Arctigenin activated caspase-3 and poly (ADP-ribose) polymerase (PARP) induced cell death. Arctigenin also inhibited the proliferation of FaDu cells by the suppression of p38, NF-κB, and Akt signaling pathways. These results suggest that Arctigenin may inhibit cell proliferation and induce apoptotic cell death in FaDu human pharyngeal carcinoma cells through both the mitochondria-mediated intrinsic pathway and the death receptor-mediated extrinsic pathway.

• Asian-Australasian Journal of Animal Sciences
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• 제29권1호
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• pp.36-42
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• 2016

Milk-related traits (milk yield, fat and protein) have been crucial to selection of Holstein. It is essential to find the current selection trends of Holstein. Despite this, uncovering the current trends of selection have been ignored in previous studies. We suggest a new formula to detect the current selection trends based on single nucleotide polymorphisms (SNP). This suggestion is based on the best linear unbiased prediction (BLUP) and the Fisher's fundamental theorem of natural selection both of which are trait-dependent. Fisher's theorem links the additive genetic variance to the selection coefficient. For Holstein milk production traits, we estimated the additive genetic variance using SNP effect from BLUP and selection coefficients based on genetic variance to search highly selective SNPs. Through these processes, we identified significantly selective SNPs. The number of genes containing highly selective SNPs with p-value <0.01 (nearly top 1% SNPs) in all traits and p-value <0.001 (nearly top 0.1%) in any traits was 14. They are phosphodiesterase 4B (PDE4B), serine/threonine kinase 40 (STK40), collagen, type XI, alpha 1 (COL11A1), ephrin-A1 (EFNA1), netrin 4 (NTN4), neuron specific gene family member 1 (NSG1), estrogen receptor 1 (ESR1), neurexin 3 (NRXN3), spectrin, beta, non-erythrocytic 1 (SPTBN1), ADP-ribosylation factor interacting protein 1 (ARFIP1), mutL homolog 1 (MLH1), transmembrane channel-like 7 (TMC7), carboxypeptidase X, member 2 (CPXM2) and ADAM metallopeptidase domain 12 (ADAM12). These genes may be important for future artificial selection trends. Also, we found that the SNP effect predicted from BLUP was the key factor to determine the expected current selection coefficient of SNP. Under Hardy-Weinberg equilibrium of SNP markers in current generation, the selection coefficient is equivalent to $2^*SNP$ effect.

• Clinical and Experimental Pediatrics
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• 제52권7호
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• pp.811-817
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• 2009

목 적 : 소아 천식에서 흡입용 스테로이드의 장기간 사용이 골밀도와 골대사에 미치는 영향에 대해 알아보고자 하였다. 방 법 : 고려대학교 안암병원 소아청소년과에서 천식으로 진단되어 최소 6개월 이상 ICS를 사용 중인 만 6세에서 12세 사이의 환아(ICS군: 26명)를 대상으로 하였다. 대조군으로 연령 및 성별이 일치하면서 LTRA만을 사용한 천식 환아(LTRA군: 15명)와 골밀도에 영향을 미칠 만한 기저 질환이 없는 정상 소아(정상군: 30명)를 선정하여 이중 에너지 X-선 흡수법으로 요추와 대퇴골 상부에서 골밀도를 측정하고, 골대사에 대한 영향은 혈청 골-특이 알칼리인산효소(BALP)를 측정하여 평가하였다. 결 과 : ICS군의 요추 골밀도는 $0.57{\pm}0.07g/cm^2$으로 LTRA군($0.55{\pm}0.06g/cm^2$) 및 정상군($0.58{\pm}0.07g/cm^2$)과 유의한 차이를 보이지 않았고(P=0.254), 대퇴골 상부의 골밀도도 ICS군에서 $0.70{\pm}0.07g/cm^2$로 LTRA군($0.66{\pm}0.06g/cm^2$)및 정상군($0.70{\pm}0.07g/cm^2$)과 유의한 차이가 없었다(P=0.297). 골밀도 Z-점수 또한 세 군에서 유의한 차이를 보이지 않았다(요추: ICS군 $-0.22{\pm}0.78$ vs. LTRA군 $-0.16{\pm}0.47$ vs. 정상군 $-0.19{\pm}0.83$, P=0.963; 대퇴골 상부: ICS 군 $-0.12{\pm}0.93$ vs. LTRA군 $0.16{\pm}0.59$ vs. 정상군 $-0.04{\pm}0.67$, P=0.560). 혈청 BALP 농도는 ICS군과 LTRA군에서 정상군에 비해 유의하게 높은 소견을 보였다(P=0.021). ICS군에서 ICS의 최근 6개월간 사용량, 총 축적 용량, 사용 기간, 투여 시작 연령과 골밀도 Z-점수는 유의한 상관관계를 보이지 않았다(모두 P>0.05). 결 론 : 천식 환아에서 저용량의 ICS 사용은 골밀도에 유의한 영향을 미치지 않음을 확인하여, ICS의 부작용에 대한 과도한 우려를 막고 치료 순응도를 높이는데 도움이 될 것으로 생각한다.