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사람의 성산자극 호르몬 α-Subunit 유전자와 결합하는 Trans-Acting Factor의 확인 및 분석
송석민,백상기,김균언 충남대학교 생물공학연구소 1999 생물공학연구지 Vol.7 No.-
The 5' flanking region of the α-subunit gene of human chorionic gonadotropin was tested for the binding with the nuclear extracts of the JAR choriocarcinoma cells. Gel mobility shift assay has demonstrated that two BstNI restriction fragments, 150 bp and 340 bp respectively, have binding activities with the nuclear extracts. The 150 bp fragment was chosen to examine in detail, since several transcription factor binding sites have already been reported within the 340 bp fragment. Band competition assay showed that the binding of 150 bp fragment is highly sequence-specific. The binding activity was further narrowed down to a 55 bp fragment, which was generated by Nsi1 digestion of the 150 bp fragment. DNasel footprinting assay revealed the binding sites on the nucleotide levels from -511 to -499. The binding sites were again confirmed by a oligonucleotide-directed mutagenesis of the 150 bp fragment followed by a gel mobility shift assay. Functional aspect of this binding was evaluated with the DNA-mediated gene transfer techniques. However, in several appoaches tried so far, no effect on the transcription efficiency was observed, implicating a structural role of the binding. In this line, it is worth to note that the binding activity of the 150 bp fragment was observed in every cell or tissue nuclear extracts tested so far.
Seok Bean Song,Nguyen Huu Tung,Tran Hong Quang,Nguyen Thi Thanh Ngan,Kyoon Eon Kim,Young Ho Kim 고려인삼학회 2012 Journal of Ginseng Research Vol.36 No.2
Panax ginseng (PG) is a globally utilized medicinal herb. The medicinal effects of PG are primarily attributable to ginsenosides located in the root and leaf. The leaves of PG are known to be rich in various bioactive ginsenosides, and the therapeutic effects of ginseng extract and ginsenosides have been associated with immunomodulatory and anti-infl ammatory activities. We examined the effect of PG leaf extract and the isolated ginsenosides, on nuclear factor (NF)-κB transcriptional activity and target gene expression by applying a luciferase assay and reverse transcription polymerase chain reaction in tumor necrosis factor (TNF)-α-treated hepatocarcinoma HepG2 cells. Air-dried PG leaf extract inhibited TNF-α-induced NF-κB transcription activity and NF-κB-dependent cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) gene expression more effi ciently than the steamed extract. Of the 10 ginsenosides isolated from PG leaves, Rd and Km most signifi cantly inhibited activity in a dose-dependent manner, with IC50 values of 12.05±0.82 and 8.84±0.99 μM, respectively. Furthermore, the ginsenosides Rd and Km inhibited the TNF-α-induced expression levels of the COX-2 and iNOS gene in HepG2 cells. Air-dried leaf extracts and their chemical components, ginsenoside Rd and Km, are involved in the suppression of TNF-α-induced NF-κB activation and NF-κB-dependent iNOS and COX-2 gene expression. Consequently, air-dried leaf extract from PG, and the purifi ed ginsenosides, have therapeutic potential as anti-infl ammatory.
Song, Seok-Bean,Tung, Nguyen Huu,Quang, Tran Hong,Ngan, Nguyen Thi Thanh,Kim, Kyoon-Eon,Kim, Young-Ho The Korean Society of Ginseng 2012 Journal of Ginseng Research Vol.36 No.2
Panax ginseng (PG) is a globally utilized medicinal herb. The medicinal effects of PG are primarily attributable to ginsenosides located in the root and leaf. The leaves of PG are known to be rich in various bioactive ginsenosides, and the therapeutic effects of ginseng extract and ginsenosides have been associated with immunomodulatory and anti-inflammatory activities. We examined the effect of PG leaf extract and the isolated ginsenosides, on nuclear factor (NF)-${\kappa}B$transcriptional activity and target gene expression by applying a luciferase assay and reverse transcription polymerase chain reaction in tumor necrosis factor (TNF)-${\alpha}$-treated hepatocarcinoma HepG2 cells. Air-dried PG leaf extract inhibited TNF-${\alpha}$-induced NF-${\kappa}B$transcription activity and NF-${\kappa}B$-dependent cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) gene expression more efficiently than the steamed extract. Of the 10 ginsenosides isolated from PG leaves, Rd and Km most significantly inhibited activity in a dose-dependent manner, with $IC_{50}$ values of $12.05{\pm}0.82$ and $8.84{\pm}0.99\;{\mu}M$, respectively. Furthermore, the ginsenosides Rd and Km inhibited the TNF-${\alpha}$-induced expression levels of the COX-2 and iNOS gene in HepG2 cells. Air-dried leaf extracts and their chemical components, ginsenoside Rd and Km, are involved in the suppression of TNF-${\alpha}$-induced NF-${\kappa}B$ activation and NF-${\kappa}B$-dependent iNOS and COX-2 gene expression. Consequently, air-dried leaf extract from PG, and the purified ginsenosides, have therapeutic potential as anti-inflammatory.
LIM domain과 상호작용 하는 transcription cofactor hTAiL-1의 기능 분석
박경숙,송석빈,김균언 충남대학교 자연과학연구소 1999 忠南科學硏究誌 Vol.26 No.1
The LIM homeodomain (LIM-HD) proteins, which contain two tandem LIM domains followed by a homeodomain, are critical transcription factors for embryonic development. The LIM domain is a conserved cysteine-rich zinc-binding motif and mediate protein-protein interactions. We have isolated two transcription activator interacting with LIM domain (hTAiL-1 and -2) from human brain cDNA library on the basis of its ability to interact with the LIM-HD protein Lh-2 (or Lhx-2). Here we report on the function of the hTAiL-1 protein. Northern blot analysis using human tissue blot revealed that hTAiL-1 is very abundantly expressed in the heart and muscle. In order to confirm that hTAL-1 interacts with LIM domain, β-galactosidase assay using yeast two-hybrid system was firstly employed, followed by an in vitro protein interaction assay and a mammalian two-hybrid assay. The results of these assay demonstrated that hTAiL-1 interact with LIM domain in a very weak mode. In vivo functional data using transfection experiments in SL-2 cells suggested that Sp1 transcription factor may be involved in the interaction of hTAiL-1 with Lh-2. In fact, hTAiL-1 stimulates the promoter activity deriven by Sp1 more efficiently than by Lh-2. In addition, effect of hTAiL-1 can be observed in a series of transfection experiment with differentiated L6 cells. Taken together, these results indicated that hTAiL-1 may mediate specific protein-protein interactions between LIM-HD and additional Sp1 basal transcription factor.
( Kyoung Won Cho ),( Seok Bean Song ),( Nguyen Huu Tung ),( Kyoon Eon Kim ),( Young Ho Kim ) 한국응용약물학회 2014 Biomolecules & Therapeutics(구 응용약물학회지) Vol.22 No.1
Panax ginseng is a medicinal herb that is used worldwide. Its medicinal effects are primarily attributable to ginsenosides located in the root, leaf, seed, and flower. The flower buds of Panax ginseng (FBPG) are rich in various bioactive ginsenosides, which exert immunomodulatory and anti-inflammatory activities. The aim of the present study was to assess the effect of 18 ginsenosides isolated from steamed FBPG on the transcriptional activity of NF-kB and the expression of tumor necrosis factor-a (TNF-α)- stimulated target genes in liver-derived cell lines. Noticeably, the ginsenosides Rk3 and Rs4 exerted the strongest activity, inhibiting NF-kB in a dose-dependent manner. SF and Rg6 also showed moderately inhibitory effects. Furthermore, these four compounds inhibited the TNF-a-induced expression of IL8, CXCL1, iNOS, and ICAM1 genes. Consequently, ginsenosides purified from steamed FBPG have therapeutic potential in TNF-a-mediated diseases such as chronic hepatic inflammation.
Tran Hong Quang,Seok Bean Song,Bui Thi Thuy Luyen,Nguyen Phuong Thao,BUIHUU TAI,Nguyen Xuan Nhiem,Phan Van Kiem,Chau Van Minh,Nguyen Thi Thanh Ngan,김영호 대한화학회 2011 Bulletin of the Korean Chemical Society Vol.32 No.11
A new compound, kalopanaxin F (3), and 11 known compounds (1, 2, 4-12), were isolated from the stem bark of Kalopanax pictus. Their structures were elucidated on the basis of chemical and spectroscopic methods. Five of the compounds (2, 3, 5, 6, and 12) significantly inhibited TNFα-induced NF-κB transcriptional activity in HepG2 cells in a dose-dependent manner, with IC50 values ranging from 6.2 to 9.1 μM. Furthermore, the transcriptional inhibitory function of these compounds was confirmed based on decreases in COX-2 and iNOS gene expression in HepG2 cells. Compounds 3-7, 9, and 12 significantly activated the transcriptional activity of PPARs dose-dependently, with EC50 values ranging from 4.1-12.7 μM. Compounds 4 and 5 exhibited PPARα, PPARγ, and PPARβ(δ) transactivational activities in a dose-dependent manner, with EC50 values of 16.0 and 17.0, 8.7 and 16.5, 26.2 and 26.3 μM, respectively.