Triptolide suppresses interleukin-1beta-induced human beta-defensin-2 mRNA expression through inhibition of transcriptional activation of NF-kappaB in A549 cells.

Jang, Byeong-Churl,Lim, Ki-Jo,Choi, In-Hak,Suh, Min-Ho,Park, Jong-Gu,Mun, Kyo-Chul,Bae, Jae-Hoon,Shin, Dong-Hoon,Suh, Seong-Il D.A. Spandidos 2007 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE Vol.19 No.5

<P>The immunosuppressive effect of triptolide has been associated with suppression of T-cell activation. However, the immunosuppressive effects of triptolide on innate immunity in the epithelial barrier remain to be elucidated. Human beta-defensin (HBD)-2 is an inducible antimicrobial peptide and plays an important role in the innate immunity. We have previously demonstrated that IL-1beta induced HBD-2 mRNA expression in A549 cells through activation of nuclear factor-kappaB (NF-kappaB) transcriptional factor as well as p38 mitogen-activated protein kinase (MAPK), c-Jun N-terminal kinase (JNK), or phosphatidylinositol-3-kinase (PI3K). In this study, we investigated effects of triptolide on IL-1beta-induced HBD-2 mRNA expression in A549 cells. Triptolide inhibited IL-1beta-induced HBD-2 mRNA expression in a dose-dependent manner. Addition of triptolide did not suppress activation of p38 MAPK, JNK, or PI3K in response to IL-1beta. Triptolide inhibited IL-1beta-induced MAPK phosphatase-1 expression at the transcriptional level and resulted in sustained phosphorylation of JNK or p38 MAPK, explaining the little effect of triptolide on IL-1beta-induced phosphorylation of these kinases. Although triptolide partially suppressed IL-1beta-mediated degradation of IkappaB-alpha and nuclear translocation of p65 NF-kappaB, triptolide potently inhibited NF-kappaB promoter-driven luciferase activity in A549 cells. These results collectively suggest that the inhibitory effect of triptolide on IL-1beta-induced HBD-2 mRNA expression in A549 cells seems to be at least in part mediated through nuclear inhibition of NF-kappaB transcriptional activity, but not inhibition of p38 MAPK, JNK, or PI3K. This inhibition may explain the ability of triptolide to diminish innate immune response.</P>

PM2.5 induces skin cell toxicity and inflammation: Inhibitory effects of ginsenoside Rg3 (20S)

Byeong-Churl Jang 한국환경생물학회 2020 한국환경생물학회 학술대회 Vol.2020 No.07

Inhibitory Effect of Dihydroartemisinin, An Active Ingredient of Artemisia annua, on Lipid Accumulation in Differentiating 3T3-L1 Preadipocytes

Jang, Byeong-Churl The Society of Korean Medicine for obesity Researc 2020 한방비만학회지 Vol.20 No.1

Objectives: Artemisinin and its derivatives extracted from Artemisia annua, a Chinese herbal medicine, have variable biological effects due to structural differences. Up to date, the anti-obesity effect of dihydroartemisinin (DHA), a derivative of artemisinin, is unknown. The purpose of this study was to investigate the anti-adipogenic and lipolytic effects of DHA on 3T3-L1 preadipocytes. Methods: Oil Red O staining and AdipoRed assay were used to measure lipid accumulation and triglyceride (TG) content in 3T3-L1 cells, respectively. Cell count analysis was used to determine the cytotoxicity of 3T3-L1 cells. Western blot and real-time reverse transcription polymerase chain reaction analyses were used to analyze the expression of protein and mRNA in 3T3-L1 cells, respectively. Results: DHA at 5 μM markedly inhibited lipid accumulation and reduced TG content in differentiating 3T3-L1 cells with no cytotoxicity. Furthermore, DHA at 5 μM inhibited the expression of CCAAT/enhancer-binding protein-α (C/EBP-α), peroxisome proliferator-activated receptor-γ (PPAR-γ), fatty acid synthase (FAS), and perilipin A as well as the phosphorylation of signal transducer and activator of transcription-3 (STAT-3) in differentiating 3T3-L1 cells. Moreover, while DHA at 5 μM had no effect on the mRNA expression of adiponectin, it strongly suppressed that of leptin in differentiating 3T3-L1 cells. However, DHA at 5 μM had no lipolytic effect on differentiated 3T3-L1 cells, as assessed by no enhancement of glycerol release. Conclusions: These results demonstrate that DHA at 5 μM has a strong anti-adipogenic effect on differentiating 3T3-L1 cells through the reduced expression and phosphorylation of C/EBP-α, PPAR-γ, FAS, perilipin A, and STAT-3.

Noni Fruit Juice의 3T3-L1 지방전구세포 분화 억제 및 인터루킨-1β 유도 Inducible Nitric Oxide Synthase 염증유전자 발현 감소 효과

장병철(Byeong-Churl Jang ) 한방비만학회 2023 한방비만학회지 Vol.23 No.1

Suppression of Lipid Accumulation in Differentiating 3T3-L1 Preadipocytes by a Standardized Commercial Juknyeok

장병철(Byeong-Churl Jang) 한방비만학회 2022 한방비만학회지 Vol.22 No.1

Objectives: Juknyeok (JN) is natural liquor extracted from bamboo stems (Phyllostachys bambusoides) and has been used as a traditional Korean medicine for improving vascular function, blood glucose, and treating stroke. Until now, the JN’s lipid-lowering effect and underlying mechanism in adipocytes are poorly understood. The aim of this study was to scrutinize the effect of a standardized commercial JN on lipid accumulation during the differentiation of 3T3-L1 preadipocytes. Methods: Lipid and triglyceride (TG) accumulation in differentiating 3T3-L1 preadipocytes were measured by Oil Red O staining and AdipoRed assay, respectively. Cell count analysis was used to ascertain 3T3-L1 cytotoxicity. Immunoblotting and Reverse transcription polymerase chain reaction analysis were used to assess protein and messenger RNA (mRNA) expression levels in 3T3-L1 cells, respectively. Results: Treatment with JN at 25 μl/ml after pH calibration with 6.35 significantly reduced lipid and TG accumulation in differentiating 3T3-L1 preadipocytes without significant cytotoxicity. On mechanistic levels, JN markedly suppressed protein expression levels of CCAAT/enhancer-binding protein (C/EBP)-β and fatty acid synthase (FAS) during the differentiation of 3T3-L1 preadipocytes. However, JN did not affect the protein expression levels of C/EBP-α, peroxisome proliferator-activated receptor-β/γ, and phosphorylation levels of signal transducer and activator of transcription-3/5 in differentiating 3T3-L1 preadipocytes. JN also reduced leptin mRNA expression levels in differentiating 3T3-L1 preadipocytes. Conclusions: JN at 25 μl/ml lowers lipid accumulation and TG content in differentiating 3T3-L1 cells, mediated through the reduced expression levels of C/EBP-β and FAS.

BAI, a novel Cdk inhibitor, enhances farnesyltransferase inhibitor LB42708-mediated apoptosis in renal carcinoma cells through the downregulation of Bcl-2 and c-FLIP?(L).

Jang, Ji Hoon,Cho, Yoon Chul,Kim, Ki Ho,Lee, Kyung Seop,Lee, Jinho,Kim, Dong Eun,Park, Jun-Soo,Jang, Byeong-Churl,Kim, Shin,Kwon, Taeg Kyu,Park, Jong-Wook Lychnia 2014 International journal of oncology Vol.45 No.4

<P>Previously, we reported the potential of a novel Cdk inhibitor, 2-[1,1'-biphenyl]-4-yl-N-[5-(1,1-dioxo-1λ6-isothiazolidin-2-yl)-1H-indazol-3-yl]acetamide (BAI) as a cancer chemotherapeutic agent. In this study, we investigated mechanisms by which BAI modulates FTI-mediated apoptosis in human renal carcinoma Caki cells. BAI synergizes with FTI to activate DEVDase, cleavage of poly ADP-ribose polymerase (PARP), and degradation of various anti-apoptotic proteins in Caki cells. BAI plus LB42708-induced apoptosis was inhibited by pretreatment with pan-caspase inhibitor, z-VAD-fmk, but not by overexpression of CrmA. The ROS scavenger, N-acetylcysteine (NAC) did not reduce BAI plus LB4270-induced apoptosis. Co-treatment of BAI and LB42708 reduced the mitochondrial membrane potential (MMP, ?ψm) in a time-dependent manner, and induced release of AIF and cytochrome?c from mitochondria in Caki cells. Furthermore, BAL plus LB42708 induced downregulation of anti-apoptotic proteins [c-FLIP?(L), c-FLIP?(s), Bcl-2, XIAP, and Mcl-1?(L)]. Especially, we found that BAI plus LB42708-induced apoptosis was significantly attenuated by overexpression of Bcl-2 and partially blocked by overexpression of c-FLIP?(L). Taken together, our results show that the activity of BAI plus LB42708 modulate multiple components in apoptotic response of human renal Caki cells, and indicate a potential as combinational therapeutic agents for preventing cancer such as renal carcinoma.</P>

Tanshinone I, an Active Ingredient of Salvia miltiorrhiza, Inhibits Differentiation of 3T3-L1 Preadipocytes and Lipid Accumulation in Zebrafish

Kwon, Hyo-Shin,Jang, Byeong-Churl The Society of Korean Medicine for obesity Researc 2020 한방비만학회지 Vol.20 No.2

Objectives: Tanshinone I is a bioactive constituent in Salvia miltiorrhiza. At present, the anti-obesity effect and mechanism of tanshinone I are not fully understood. Here we investigated the effect of tanshinone I on lipid accumulation in 3T3-L1 preadipocytes and zebrafish. Methods: Lipid accumulation and triglyceride (TG) content in 3T3-L1 cells were determined by Oil Red O staining and AdipoRed assay, respectively. The expression and phosphorylation levels of adipogenic/lipogenic proteins in 3T3-L1 cells were evaluated by Western blotting. The messenger RNA (mRNA) expression levels of adipogenic/lipogenic markers and leptin in 3T3-L1 cells were measured by reverse transcription polymerase chain reaction (RT-PCR). Lipid accumulation in zebrafish was assessed by LipidGreen2 staining. Results: Tanshinone I at 5 μM largely blocked lipid accumulation and reduced TG content in differentiating 3T3-L1 cells. Furthermore, tanshinone I decreased the expression of CCAAT/enhancer-binding protein-α (C/EBP-α), peroxisome proliferator-activated receptor-γ (PPAR-γ), fatty acid synthase (FAS), acetyl CoA carboxylase (ACC), and perilipin A but also the phosphorylation of signal transducer and activator of transcription-3 (STAT-3) in differentiating 3T3-L1 cells. In addition, tanshinone I increased the phosphorylation of adenosine 3',5'-cyclic monophosphate (cAMP)-activated protein kinase (AMPK) while decreased the intracellular adenosine triphosphate (ATP) content with no change in the phosphorylation and expression of liver kinase-B1 in differentiating 3T3-L1 cells. Importantly, tanshinone I also reduced the extent of lipid deposit formation in developing zebrafish. Conclusions: These findings demonstrate that tanshinone I has strong anti-adipogenic effects on 3T3-L1 cells and reduces adiposity in zebrafish, and these anti-adipogenic effect in 3T3-L1 cells are mediated through control of C/EBP-α, PPAR-γ, STAT-3, FAS, ACC, perilipin A, and AMPK.

Ginsenoside Rb2 suppresses cellular senescence of human dermal fibroblasts by inducing autophagy

Kyeong Eun Yang,Soo-Bin Nam,Minsu Jang,Junsoo Park,Ga-Eun Lee,Yong-Yeon Cho,Byeong-Churl Jang,Cheol-Jung Lee,Jong-Soon Choi The Korean Society of Ginseng 2023 Journal of Ginseng Research Vol.47 No.2

Background: Ginsenoside Rb2, a major active component of Panax ginseng, has various physiological activities, including anticancer and anti-inflammatory effects. However, the mechanisms underlying the rejuvenation effect of Rb2 in human skin cells have not been elucidated. Methods: We performed a senescence-associated β-galactosidase staining assay to confirm cellular senescence in human dermal fibroblasts (HDFs). The regulatory effects of Rb2 on autophagy were evaluated by analyzing the expression of autophagy marker proteins, such as microtubule-associated protein 1A/1B-light chain (LC) 3 and p62, using immunoblotting. Autophagosome and autolysosome formation was monitored using transmission electron microscopy. Autophagic flux was analyzed using tandem-labeled GFP-RFP-LC3, and lysosomal function was assessed with Lysotracker. We performed RNA sequencing to identify potential target genes related to HDF rejuvenation mediated by Rb2. To verify the functions of the target genes, we silenced them using shRNAs. Results: Rb2 decreased β-galactosidase activity and altered the expression of cell cycle regulatory proteins in senescent HDFs. Rb2 markedly induced the conversion of LC3-I to LC3-II and LC3 puncta. Moreover, Rb2 increased lysosomal function and red puncta in tandem-labeled GFP-RFP-LC3, which indicate that Rb2 promoted autophagic flux. RNA sequencing data showed that the expression of DNA damage-regulated autophagy modulator 2 (DRAM2) was induced by Rb2. In autophagy signaling, Rb2 activated the AMPK-ULK1 pathway and inactivated mTOR. DRAM2 knockdown inhibited autophagy and Rb2-restored cellular senescence. Conclusion: Rb2 reverses cellular senescence by activating autophagy via the AMPK-mTOR pathway and induction of DRAM2, suggesting that Rb2 might have potential value as an antiaging agent.

동아 추출물과 프로바이오틱 유산균의 생리활성 및 과민성대장증후군 개선 효과

안용근 ( Yong Geun Ann ),장병철 ( Byeong Churl Jang ),박세준 ( Se Joon Park ) 한국식품영양학회 2013 韓國食品營養學會誌 Vol.26 No.1

동아(冬瓜, wax gourd, Benincase hispida) 추출물과 프로바이오틱 유산균(Lactobacillus casei와 Bifidobacterium bifidum) (이하 유산균이라 한다)의 생리활성을 분석하였다. 동아 추출물은 안기오텐신전환효소의 활성을 47.9%, 티로시나제의 활성을 13.2% 저해하였고, 항산화 활성을 23.4% 나타냈다. 설사 증세자 120명에게 동아 추출물 제제를 72시간 복용시킨 결과 117명이 개선되었고, 6∼12시간째에 개선율이 가장 높았다. 개선율은 80%가 치료되는 상태를 기준하였다. 유산균 제제는 안기오텐신전환효소의 활성을 21.4% 저해하였다. 설사 증세자 120명에게 유산균 제제를 72시간 복용시킨 결과 108명이 개선되었고, 24시간째에 개선율이 가장 높았다. 이들 결과를 바탕으로, 과민성대장증후군 개선을 위한 동아 추출물과 유산균을 함유한 제제(tablet)를 개발하였다. 이 제품은 안기오텐신전환효소의 활성을 27.1% 저해하였고, 항산화 활성을 20.3% 나타냈다. 인체폐암 A549 세포에 100㎍/㎖ 및 250㎍/㎖ 농도로 이 제제를 가하여 24시간 반응시킨 결과, 세포 증식을 67% 억제하였고, 동아 추출물 제제나 프로바이오틱 유산균 제제 단독으로 작용시킨 것보다 억제율이 높았다. 또 이 제제를 인체대장암 HCT116 세포에 100㎍/㎖ 농도로 가한 결과, 24시간 후 세포 증식이 70% 억제되었고, 동아 추출물 제제나 유산균 제제 단독으로 작용시킨 것보다 개선율과 개선 속도가 높았다. 이 제제를 설사형, 변비형, 일반형의 과민성대장증후군 증세자 164명에게 72시간 복용시킨 결과 100% 개선시켰으며, 개선율은 3∼6시간째에 가장 높았고, 동아 추출물 제제나 유산균 제제 단독으로 사용한 것보다 개선율과 개선속도가 높았다. Biological activities of wax gourd (Benincase hispida) extract and lactic acid bacteria (Lactobacillus casei and Bifidobacterium bifidum) were investigated in this study. Wax gourd extract reduced the activity of angiotensin-converting enzyme (ACE) by 47.9%, of tyrosinase by 13.2%, and had an anti-oxidant activity of 23.4%. Oral administration of wax gourd extract for 72 hours improved the symptom of loose bowels for 120 patients with its highest improvement rates within 6 to 12 hours. The improvement rates were standardized by the curative state by 80%. Lactic acid bacteria preparations reduced the activity of ACE by 21.49%. Oral administration of lactic acid bacteria preparations for 72 hours improved the symptom of loose bowels for 108 patients with its highest improvement rates after 24 hours. On the basis of these results, the tablets containing both wax gourd extract and lactic acid bacteria preparations for the improvement of irritable bowel syndromes were developed. The tablets reduced the activity of ACE by 27.1% and exhibited an anti-oxidant activity of 20.3%. Treatment of the tablets at 100㎍/㎖ and 250㎍/㎖ for 24 hours inhibited the growth of A549 human lung cancer cells by 67%, which was much higher than that of each wax gourd extract or lactic acid bacteria. In addition, treatment of the tablets at 100㎍/㎖ for 24 hours reduced the growth of HCT-116 human colon cancer cells by 70%. Oral administration of the tablets to the patients with loose bowels led to higher improvement rates and speed than each wax gourd extract or lactic acid bacteria. Oral administration of the tablets to the patients with irritable bowel syndromes of loose bowels, constipation, or general type for 72 hours improved their symptoms by 100% with the highest improvement rates within 3 to 6 hours. Furthermore, the improvement rates and speed by the tablets was much higher than each wax gourd extract or lactic acid bacteria.

Action mechanism of anti-wrinkle effect of Rhamnus yoshinoi methanol extract in human dermal fibroblast and keratinocyte cell lines

Hyun Ok Kim,Kyoung Ran Shin,Byeong-Churl Jang,Young Chul Kim 한국독성학회 2020 Toxicological Research Vol.36 No.1

Rhamnus yoshinoi is a deciduous broad-leaf bush and endemic species widely found in Korea. Recently, we reported that R. yoshinoi methanol extract (RYME) had excellent antioxidant activity and inhibition of collagenase and elastase activity in cell-free system. In this study, we investigated the ability of RYME to control the mRNA and protein expression levels of the known skin wrinkle-related factors in cultured human dermal fibroblast and keratinocyte cell lines. Treatment with 100 or 200 μg/mL RYME strongly blocked the UVB-induced downregulation of type 1 collagen mRNA expression (p < 0.001) and partially blocked the UVB-induced upregulation of MMP-3 mRNA expression in HaCaT human keratinocytes (p < 0.05 or p < 0.001). Treatment with RYME at 100 μg/mL considerably decreased MMP-1 mRNA expression in UVB-exposed HaCaT cells (p < 0.01). In HaCaT cells, RYME exhibited the potential to improve UV light-induced skin wrinkles. Moreover, RYME selectively inhibited the UVB-induced ERK-1/2 protein phosphorylation in CCD-986sk human dermal fibroblasts at 80 and 160 μg/mL. UV-induced ERK-1/2 protein phosphorylation is one of the major mechanisms of the generation of UV-induced skin wrinkles. Therefore, it is likely that the anti-skin wrinkling effect of RYME could be attributable to selective inhibition of UV induced ERK-1/2 protein phosphorylation.