자초(紫草) 열수추출물이 각질형성세포 HaCaT의 세포 연접 관련 유전자의 발현에 미치는 영향 연구

조남준 ( Namjoon Cho ),이병권 ( Byeongkwon Lee ),이웅희 ( Woonghee Lee ),김기광 ( Keekwang Kim ),김균언 ( Kyoon Eon Kim ),한효상 ( Hyosang Han ) 대한본초학회 2017 大韓本草學會誌 Vol.32 No.3

Objectives : The aim of this research was to determine the diverse effects of Lithospermi Radix Water Extract (LR) on human keratinocyte HaCaT cells, and to examine whether those effects could be applied to the human skin. Methods : We examined effect of LR on the cell viability of using the MTS assay in human keratinocyte HaCaT cells. The antioxidation effect of LR was analyzed relative to the well-known antioxidant resveratrol, using an ABTS assay. Quantitative RT-PCR analysis revealed that, in HaCaT cells, LR influenced the mRNA expression of tight-junction genes associated with skin moisturization. Furthermore, a wound-healing assay demonstrated altered cell migration in LR-treated HaCaT cells. Result : The cytotoxicity was confirmed to be higher in LR at a concentration of 800 ㎍/㎖ using the MTS assay in HaCaT cells. In comparison to 100 μM resveratrol, 1,600 ㎍/㎖ LR showed either a similar or superior antioxidation effect. LR treatment in HaCaT cells reduced the mRNA expression levels of claudin 3, claudin 4, claudin 6, claudin 8, and ZO-2 to less than 0.80-fold, whereas JAM-A and Tricellulin mRNA expression level increased more than 1.33- fold. In addition, HaCaT cells migration was decreased to 83.9% by LR treatment. Conclusions : LR of antioxidation activity will have an anti-aging effect on the skin by reducing oxidative stress. Further studies are required to address the implications for human skin, given LR`s effects of altering mRNA expression of tight junction-related gene and decreasing cell migration of HaCaT cells.

Cudrania tricuspidata leaf extracts and its components, chlorogenic acid, kaempferol, and quercetin, increase claudin 1 expression in human keratinocytes, enhancing intercellular tight junction capacity

김재환,Cho Namjoon,Kim Eun-Mi,Park Ki-Sun,Kang Yeon Woo,Nam Joong Hyeon,Nam Myoung Soo,Kim Kee K. 한국응용생명화학회 2020 Applied Biological Chemistry (Appl Biol Chem) Vol.63 No.3

Dysfunction of tight junctions and their components can cause diverse skin diseases. Here, we investigated the expression of claudin 1, a major tight junction protein, and changes of tight junction capacity upon treatment of the extracts of Cudrania tricuspidata (C. tricuspidata) and its components, chlorogenic acid, kaempferol, and quercetin. The effects of ethanol extracts of C. tricuspidata (EECT) and water extracts of C. tricuspidata (WECT) on the viability of human keratinocyte HaCaT cells were assessed by cell proliferation assay. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was conducted to measure the expression of claudin 1 mRNA. The protein expression of claudin 1 was analyzed by western blot and its tight junctional distribution was observed with immunofluorescence microscopy analysis. The tight junction capacity was analyzed by dispase assay. Upon treatment of WECT to HaCaT cells, the mRNA and protein expressions of claudin 1 were increased. In addition, chlorogenic acid, kaempferol, and quercetin increased claudin 1 protein expression levels in a dose-dependent manner. WECT and these three compounds enhanced the tight junction capacity of HaCaT cells in dispase assay. WECT, and its components, such as chlorogenic acid, kaempferol, and quercetin, upregulates both mRNA and protein expressions of claudin 1, which leads to the enhancement of tight junction capacity. Thus, WECT could be a therapeutic approach for treating tight junction-disrupted conditions such as atopic dermatitis and psoriasis.

The implications of alternative pre-mRNA splicing in cell signal transduction

Choi Sunkyung,Cho Namjoon,Kim Kee K. 생화학분자생물학회 2023 Experimental and molecular medicine Vol.55 No.-

Cells produce multiple mRNAs through alternative splicing, which ensures proteome diversity. Because most human genes undergo alternative splicing, key components of signal transduction pathways are no exception. Cells regulate various signal transduction pathways, including those associated with cell proliferation, development, differentiation, migration, and apoptosis. Since proteins produced through alternative splicing can exhibit diverse biological functions, splicing regulatory mechanisms affect all signal transduction pathways. Studies have demonstrated that proteins generated by the selective combination of exons encoding important domains can enhance or attenuate signal transduction and can stably and precisely regulate various signal transduction pathways. However, aberrant splicing regulation via genetic mutation or abnormal expression of splicing factors negatively affects signal transduction pathways and is associated with the onset and progression of various diseases, including cancer. In this review, we describe the effects of alternative splicing regulation on major signal transduction pathways and highlight the significance of alternative splicing.

Intron retention decreases METTL3 expression by inhibiting mRNA export to the cytoplasm

Sangsoo Lee,정해수,최선경,Namjoon Cho,Eun-Mi Kim,김기광 생화학분자생물학회 2023 BMB Reports Vol.56 No.9

Methyltransferase-like 3 (METTL3), a key component of the m6Amethyltransferase complex, regulates the splicing, nuclear transport,stability, and translation of its target genes. However, themechanism underlying the regulation of METTL3 expressionby alternative splicing (AS) remains unknown. We analyzedthe expression pattern of METTL3 after AS in human tissues andconfirmed the expression of an isoform retaining introns 8 and9 (METTL3-IR). We confirmed the different intracellular localizationsof METTL3-IR and METTL3 proteins using immunofluorescencemicroscopy. Furthermore, the endogenous expressionof METTL3-IR at the protein level was different from that at themRNA level. We found that 3’-UTR generation by intron retention(IR) inhibited the export of METTL3-IR mRNA to thecytoplasm, which in turn suppressed protein expression. Tothe best of our knowledge, this is the first study to confirm theregulation of METTL3 gene expression by AS, providing evidencethat the suppression of METTL3 protein expression byIR is an integral part of the mechanism by which 3’-UTR generationregulates protein expression via inhibition of RNA exportto the cytoplasm.

맥문동 열수 추출물이 Poly I:C를 처리한 폐암세포주의 사멸 및 염증성 사이토카인 발현에 미치는 영향

강다연 ( Dayeon Kang ),조남준 ( Namjoon Cho ),렌친핸드 ( Gereltuya Renchinkhand ),이보희 ( Bo-hee Lee ),김은미 ( Eun-mi Kim ),남명수 ( Myoung Soo Nam ),김기광 ( Keekwang Kim ) 대한본초학회 2021 大韓本草學會誌 Vol.36 No.1

Objectives : Virus infection through the respiratory tract causes various inflammatory diseases such as pneumonia, cystic fibrosis, and obstructive pulmonary disease, causing enormous social damage. Therefore, it is very important to develop a treatment and prevention of infectious diseases. In this study, we investigated the effect of water extracts of Liriope muscar i (WELM), known to improve lung function, on the inflammatory response of lung carcinoma cell line A549 cells induced by the viral double stranded RNA mimetic Polyinosinic:polycytidylic acid (Poly I:C). Methods : The cell viability by WELM treatment was analyzed using MTS assay in A549 cells. After inducing an inflammatory response to WELM-treated A549 cells with Poly I:C, the degree of apoptosis was confirmed through bright field microscopy. Interferon beta (IFN-β) mRNA expression level in A549 cells was analyzed by quantitative reverse transcription PCR (qRT-PCR). Results : WELM treatment has no significant effect on cell viability of A549 cells. We confirmed that pre-treatment of WELM effectively reduces the Poly I:C-induced apoptotic cell death in A549 cells. In addition, it was confirmed that the mRNA expression level of IFN-β, a pro-inflammatory cytokine increased by Poly I:C treatment, was significantly suppressed by WELM treatment in A549 cells. Conclusions : These results provide the evidence that WELM is effective at inhibiting inflammation on respiratory viral infections and suggest that Liriope muscari might be a valuable natural substance in the prevention and treatment of infectious diseases.

발모 향상을 위한 한약재의 개발 및 효과 확인

최선경(Sunkyung Choi),조남준(Namjoon Cho),김기광(Kee K. Kim) 한국생물공학회 2016 KSBB Journal Vol.31 No.4

Hair loss affects both men and women of all ages and often significantly affects social and psychologic health. Recent therapeutic approach for hair loss such as finasteride and minoxidil focused on regulation of hormonal system blood flow. However, long-term use of these drugs caused adverse effects. To develop herbal medicine for therapeutic effect on hair growth, here we screened the 10 medicinal herbs (Red ginseng, Licorice, Ulmus, Barberry root, Lycium root, Rehmanniae radix crudus, Sophora root, Sweet flag, Polygala root, Achyranthes) based on oriental medicine literature. We measured cytotoxicity, anti-oxidant activity, and 5-α reductase inhibitory effect of the herbal medicine on human dermal papilla (DP) cells to investigate therapeutic effect of the herbal medicine. Treatment of the 1% herbal medicine did not show any cytotoxic effects, and cell growth was increased by treatment of the 0.1% herbal medicine. In addition, the herbal medicine showed stronger antioxidant activity than resveratrol and comparable inhibitory activity of 5-α reductase with finasteride. Furthermore, when applied to in vivo mouse model, we also observed increases in the number and length of hair of the herbal medicine-treated group. These results suggest that the herbal medicine promotes hair growth by its antioxidant activity and inhibitory activity of 5-α reductase and might therefore be a promising hair growth-promoting agent.

꽃송이버섯 추출물의 간세포에서 대사 활성 효과

백소정(So-Jeong Baek),조남준(Namjoon Cho),조성진(Sung-Jin Cho),김은미(Eun-Mi Kim),김기광(Kee Kwang Kim) 한국생물공학회 2020 KSBB Journal Vol.35 No.2

Sulfonylurea treatment is used for diabetes mellitus, a metabolic disease caused by abnormal absorption of glucose in the body. But it is accompanied by side effects in vivo when taken for a long time. Therefore, there is a need for research on natural materials having similar effects to these therapeutic agents and having fewer side effects that can effectively regulate blood sugar. In this study, we investigated the antioxidative efficacy of Sparassis crispa (S. crispa) extract and the activity of ATP, glycolysis, and mitochondrial membrane potential using human hepatocyte HepG2 cells. As a result, S. crispa extract showed high antioxidant activity and induced a significant increase in HepG2 cell activity and ATP production. S. crispa extract also significantly accelerated the process and mitochondrial membrane potential. Taken together, these results suggest that S. crispa extract has the potential to be developed as a natural material for improving diabetic and diabetic complications by activating hepatic energy metabolism.

Dexamethasone Facilitates NF-κB Signal Pathway in TNF-α Stimulated Rotator Cuff Tenocytes

( Jong-hun Ji ),( Young-yul Kim ),( Kaushal Patel ),( Namjoon Cho ),( Sang-eun Park ),( Myung-sup Ko ),( Suk-jae Park ),( Jong Ok Kim ) 한국미생물 · 생명공학회 2019 Journal of microbiology and biotechnology Vol.29 No.2

Corticosteroids are commonly used for pain control in rotator cuff tear. Deregulated NF-κB activation is a hallmark of chronic inflammatory diseases and has been responsible for the pathogenesis of rotator cuff tear. The Dexamethasone(DEXA) is a synthetic corticosteroid. The purpose of this study was to examine the exact effect of dexamethasone on NF-κB signaling in rotator cuff tear. We measured NF-κB expression in four groups: control, TNF-α-treated, DEXA-treated, and combined treatment with TNF-α and DEXA. Tenocytes were isolated from patients with rotator cuff tears and pre-incubated with TNF-α (10 ng/ml), DEXA (1 μM), or both of them for 10 min, 1 h, and 2 h. Expression of p65, p50, and p52 in the nuclei and cytosol was analyzed by western blotting and immunofluorescence imaging using confocal microscopy. We also evaluated nucleus/cytosol (N/C) ratios of p65, p50, and p52. In our study, the combined treatment with DEXA and TNF-α showed increased N/C ratios of p65, p50, and p52 compared with those in the TNF-α group at all time points. Additionally, in the DEXA group, N/C ratios of p65, p50, and p52 gradually increased from 10 min to 2 h. In conclusion, DEXA promoted the nuclear localization of p65, p50, and p52, but was not effective in inhibiting the inflammatory response of TNF-α-stimulated rotator cuff tear.

Rbfox family proteins make the homo- and hetero-oligomeric complexes

Choi, Sunkyung,Kim, Yong-Eun,Kim, Jae Whan,Cho, Namjoon,Cheon, Seonghye,Kim, Kee K. Academic Press 2018 Biochemical and biophysical research communication Vol. No.

<P><B>Abstract</B></P> <P>Rbfox family of proteins that consists of Rbfox1, Rbfox2, and Rbfox3 in mammals regulates alternative pre-mRNA splicing in various tissues via direct binding to their RNA binding element. Although many studies have indicated the splicing activity of each member of the Rbfox family, the interactions of Rbfox family proteins are largely unknown. Here, we have investigated interactions among Rbfox family proteins. Co-immunoprecipitation (Co-IP) and GST-pull down assays confirmed that Rbfox proteins form homo and hetero complexes. Moreover, <I>in vivo</I> crosslinking using disuccinimidyl suberate treatment indicated that the Rbfox proteins form a dimer which then assembles with other proteins to form a large multiprotein complex. Duolink <I>in situ</I> proximity ligation (PLA) assay revealed that neuron specific Rbfox3 protein interacts with other Rbfox family proteins. This study is the first to provide an evidence that Rbfox family proteins form homo- and hetero-oligomeric complexes <I>in vivo</I>.</P>