• Title/Summary/Keyword: Viability Mechanism

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The Molecular Mechanism of Baicalin on RANKL-induced Osteoclastogenesis in RAW264.7 Cells

  • Ko, Seon-Yle
    • International Journal of Oral Biology
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    • v.38 no.2
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    • pp.67-72
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    • 2013
  • This study examined the anti-osteoclastogenic effects of baicalin on receptor activator of NF-${\kappa}$B ligand (RANKL)-induced RAW264.7 cells. Baicalin is a flavonoid that is produced by Scutellaria baicalensis and is known to have multiple biological properties, including antibacterial, anti-inflammatory and analgesic effects. The effects of baicalin on osteoclasts were examined by measuring 1) cell viability; 2) the formation of tartrate-resistant acid phosphatase (TRAP) (+) multinucleated cells; 3) RANK/RANKL signaling pathways and 4) mRNA levels of osteoclast-associated genes. Baicalin inhibited the formation of RANKL-stimulated TRAP (+) multinucleated cells and also suppressed the RANKL-stimulated activation of p-38, ERK, cSrc and AKT signaling. Baicalin also inhibited the RANKL-stimulated degradation of $I{\kappa}B$ in RAW264.7 cells. In addition, the RANKL-stimulated induction of NFATc1 transcription factors was found to be abrogated by this flavonoid. Baicalin was further found to decrease the mRNA expression of osteoclast-associated genes, including carbonic anhydrase II, TRAP and cathepsin K in the RAW264.7 cells. Our data thus demonstrate that baicalin inhibits osteoclastogenesis by inhibiting the RANKL-induced activation of signaling molecules and transcription factors in osteoclast precursors.

Necroptosis Is a Mechanism of Death in Mouse Induced Hepatocyte-Like Cells Reprogrammed from Mouse Embryonic Fibroblasts

  • Lee, Yun-Suk;Park, Kyung-Mee;Yu, Lina;Kwak, Ho-Hyun;Na, Hee-Jun;Kang, Kyung-Sun;Woo, Heung-Myong
    • Molecules and Cells
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    • v.41 no.7
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    • pp.639-645
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    • 2018
  • Liver transplantation is recommended for patients with liver failure, but liver donors are limited. This necessitates the development of artificial livers, and hepatocytes are necessary to develop such artificial livers. Although induced hepatocyte-like cells are used in artificial livers, the characteristics of mouse induced hepatocyte-like cells (miHeps) reprogrammed with embryonic fibroblasts have not yet been clarified. Therefore, this study investigated the mechanisms underlying the survival, function, and death of miHeps. miHeps showed decreased cell viability, increased cytotoxicity, decreased hepatic function, and albumin and urea secretion at passage 14. Addition of necrostatin-1 (NEC-1) to miHeps inhibited necrosome formation and reactive oxygen species generation and increased cell survival. However, NEC-1 did not affect the hepatic function of miHeps. These results provide a basis for development of artificial livers using hepatocytes.

Protective Effects of Verapamil against H2O2-Induced Apoptosis in Human Lens Epithelial Cells

  • Wang, Zhuo;Wang, Dan;Li, Yan;Zhang, Xiuli
    • Biomolecules & Therapeutics
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    • v.22 no.6
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    • pp.553-557
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    • 2014
  • Verapamil is used in the treatment of hypertension, angina pectoris, and atrial fibrillation. Recently, several studies have demonstrated that verapamil increased the optic nerve head blood flow and improved the retrobulbar circulation. All these show that verapamil is potentially useful for ophthalmic treatment. Thus, the aim of this study is to investigate whether verapamil could protect human lens epithelial cell (HLEC) from oxidative stress induced by $H_2O_2$ and the cellular mechanism underlying this protective function. The viability of HLEC was determined by the MTT assay and apoptotic cell death was analyzed by Hoechst 33258 staining. Moreover, Caspase-3 expression was detected by immunocytochemistry and flow cytometry analysis. We also detected Caspase-3 mRNA expression by reverse-transcription-polymerase chain reaction and the GSH content in cell culture. The results showed that oxidative stress produced significant cell apoptotic death and it was reduced by previous treatment with the verapamil. Verapamil was effective in reducing HLEC death mainly through reducing the expression level of apoptosis-related proteins, caspase-3, and increasing glutathione content. Therefore, it was suggested that verapamil was effective in reducing HLEC apoptosis induced by $H_2O_2$.

Enhanced Biotransformation Productivity of Gamma-Decalactone from Ricinoleic Acid Based on the Expanded Vermiculite Delivery System

  • Guan, Shimin;Rong, Shaofeng;Wang, Mengze;Cai, Baoguo;Li, Qianqian;Zhang, Shuo
    • Journal of Microbiology and Biotechnology
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    • v.29 no.7
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    • pp.1071-1077
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    • 2019
  • Natural gamma-decalactone (GDL) produced by biotransformation is an essential food additive with a peach-like aroma. However, the difficulty of effectively controlling the concentration of the substrate ricinoleic acid (RA) in water limits the biotransformation productivity, which is a bottleneck for industrialization. In this study, expanded vermiculite (E-V) was utilized as a carrier of RA to increase its distribution in the medium. E-V and three commonly used organic compounds were compared with respect to their effects on the biotransformation process, and the mechanism was revealed. Scanning electron microscopy, Fourier transform infrared spectroscopy and thermogravimetric analysis indicated that RA was physically adsorbed onto the surface of and inside E-V instead of undergoing a chemical reaction, which increased the opportunity for interactions between microorganisms and the substrate. The highest concentration of GDL obtained in the medium with E-V was 6.2 g/l, which was 50% higher than that in the reference sample. In addition, the presence of E-V had no negative effect on the viability of the microorganisms. This study provides a new method for producing natural GDL through biotransformation on an industrial scale.

Nicorandil alleviated cardiac hypoxia/reoxygenation-induced cytotoxicity via upregulating ketone body metabolism and ACAT1 activity

  • Bai, Yan Ping;Han, Lei Sen
    • The Korean Journal of Physiology and Pharmacology
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    • v.23 no.1
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    • pp.37-45
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    • 2019
  • To study the effect of nicorandil pretreatment on ketone body metabolism and Acetyl-CoA acetyltransferase (ACAT1) activity in hypoxia/reoxygenation (H/R)-induced cardiomyocytes. In our study, we applied H9c2 cardiomyocytes cell line to evaluate the cardioprotective effects of nicorandil. We detected mitochondrial viability, cellular apoptosis, reactive oxygen species (ROS) production and calcium overloading in H9c2 cells that exposed to H/R-induced cytotoxicity. Then we evaluated whether nicorandil possibly regulated ketone body, mainly ${\beta}$-hydroxybutyrate (BHB) and acetoacetate (ACAC), metabolism by regulating ACAT1 and Succinyl-CoA:3-ketoacid coenzyme A transferase 1 (OXCT1) protein and gene expressions. Nicorandil protected H9c2 cardiomyocytes against H/R-induced cytotoxicity dose-dependently by mitochondria-mediated anti-apoptosis pathway. Nicorandil significantly decreased cellular apoptotic rate and enhanced the ratio of Bcl-2/Bax expressions. Further, nicorandil decreased the production of ROS and alleviated calcium overloading in H/R-induced H9c2 cells. In crucial, nicorandil upregulated ACAT1 and OXCT1 protein expressions and either of their gene expressions, contributing to increased production of cellular BHB and ACAC. Nicorandil alleviated cardiomyocytes H/R-induced cytotoxicity through upregulating ACAT1/OXCT1 activity and ketone body metabolism, which might be a potential mechanism for emerging study of nicorandil and other $K_{ATP}$ channel openers.

Key Success Factors of Home-Based Business among Female Entrepreneur in Saudi Arabia

  • bin Dahari, Zainurin;Abu Bakar, Abdul Rahim;Al-Gosaibi, Sahar
    • Asia Pacific Journal of Business Review
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    • v.3 no.2
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    • pp.43-66
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    • 2019
  • Home-based business (HBB) is one of the fastest growing form of business start-ups where the business is conducted from home. In develop economies, the HBB industry is an engine for economic growth that has proven its viability through significant contribution to the national GDP. In view of its importance, the Saudi Ministry of Commerce and Industry encourage local women to start and develop their own HBB as it gives them the flexibility while still contributing to the national economy. Although various initiatives have been taken place, little information is available about Saudi HBB. This study therefore aims to determine the factors that contribute to the growth as well as the challenges of female HBB in Saudi Arabia. The study was based on qualitative approach which adopted an in-depth interview with eleven Saudi females who run HBB. The findings revealed that HBB contributing factors comprised of access to funding, sufficiency of savings and the influence of the intrinsic and extrinsic motivation in starting the business. Meanwhile, the obstacles are government regulations and policies, culture, and home design. The study recommendations include improving banks and service institution policy and procedures to enable HBB to have access to funding and services and to develop legal policies to protect the right of the HBB operator and customers. Finally, the study also suggests future research on managerial factors that can contribute to HBB female success, the stress coping mechanism of HBB and the factors contributing to the difference between HBB in growth strategies.

Gomisin G Suppresses the Growth of Colon Cancer Cells by Attenuation of AKT Phosphorylation and Arrest of Cell Cycle Progression

  • Maharjan, Sony;Park, Byoung Kwon;Lee, Su In;Lim, Yoongho;Lee, Keunwook;Lee, Younghee;Kwon, Hyung-Joo
    • Biomolecules & Therapeutics
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    • v.27 no.2
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    • pp.210-215
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    • 2019
  • Colorectal cancer is one of the leading causes of cancer related death due to a poor prognosis. In this study, we investigated the effect of Gomisin G on colon cancer growth and examined the underlying mechanism of action. We found that Gomisin G significantly suppressed the viability and colony formation of LoVo cells. Gomisin G reduced the phosphorylation level of AKT implying that Gomisin G suppressed the PI3K-AKT signaling pathway. Gomisin G also induced apoptosis shown by Annexin V staining and an increased level of cleaved poly-ADP ribose polymerase (PARP) and Caspase-3 proteins. Furthermore, Gomisin G remarkably triggered the accumulation of cells at the sub-G1 phase which represents apoptotic cells. In addition, the level of cyclin D1 and phosphorylated retinoblastoma tumor suppressor protein (Rb) was also reduced by the treatment with Gomisin G thus curtailing cell cycle progression. These findings show the suppressive effect of Gomisin G by inhibiting proliferation and inducing apoptosis in LoVo cells. Taken together, these results suggest Gomisin G could be developed as a potential therapeutic compound against colon cancer.

Inhibitory Effects of Alveopora japonica Extract on Melanin Synthesis (거품돌산호 추출물의 멜라닌 합성 억제 효능)

  • Shim, Joong Hyun
    • Korean Journal of Pharmacognosy
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    • v.52 no.3
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    • pp.143-148
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    • 2021
  • This study was performed to elucidate the inhibitory effects of Alveopora japonica extract on melanin synthesis by measuring the levels of cell viability, mRNA expression, tyrosinase activity, and melanin production in the B16F10 cell line. The effects of A. japonica extract on tyrosinase-related protein 1 (TYRP1), TYRP2, tyrosinase (TYR), and microphthalmia-associated transcription factor (MITF) mRNA expression levels and melanin content were determined. Quantitative real-time RT-PCR show that A. japonica extract decrease the mRNA expression levels of TYRP1, TYRP2, TYR, and MITF in B16F10 cell line, resulting in lower levels of melanin production compared to α-MSH-treated B16F10 cells. Tyrosinase activity assays reveal that A. japonica extract decrease melanin production in B16F10 cells. These results demonstrate the whitening effects of A. japonica extract on B16F10 cells; thus, A. japonica extract is a potent ingredient for skin whitening. Further research is needed on the mechanism of action of A. japonica extract. Such research will benefit not only cosmetics, but also the health food and medical industries.

Fluoxetine affects cytosolic cAMP, ATP, Ca2+ responses to forskolin, and survival of human ovarian granulosa tumor COV434 cells

  • Nguyen, Thi Mong Diep;Klett, Daniele;Combarnous, Yves
    • The Korean Journal of Physiology and Pharmacology
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    • v.25 no.3
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    • pp.189-195
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    • 2021
  • Fluoxetine (FLX), a selective serotonin reuptake inhibitor antidepressant, exhibits various other mechanisms of action in numerous cell types and has been shown to induce cell death in cancer cells, paving the way for its potential use in cancer therapy. The aim of this study was to determine the off-target effects of the anti-depressant drug FLX, on the human ovarian granulosa tumor COV434 cells stimulated by forskolin (FSK), by measuring the real-time kinetics of intracellular cyclic AMP (cAMP), ATP level, cytoplasmic calcium ([Ca2+]cyt) and survival of COV434 cells. We show that incubating COV434 cells with FLX (between 0.6 and 10 μM) induces a decrease in intracellular cAMP response to FSK, a drop in ATP content and stimulates cytoplasmic Ca2+ accumulation in COV434 cells. Only the highest concentrations of FLX (5-10 μM) diminished cell viability. The present report is the first to identify an action mechanism of FLX in human tumor ovarian cells COV434 cells and thus opening the way to potential use of fluoxetine as a complementary tool, in granulosa tumor treatments.

Whitening Effects of Anthricin on B16F10 Cells (B16F10 세포에서 Anthricin의 미백 효능)

  • Shim, Joong Hyun
    • Korean Journal of Pharmacognosy
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    • v.52 no.1
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    • pp.13-18
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    • 2021
  • This study was performed to clarify the whitening effects of anthricin on the B16F10 cell line. In order to elucidate the whitening effects of anthricin on the B16F10 cell line, cell viability, messenger ribonucleic acid (mRNA) expressions, tyrosinase activity assay, and melanin production assay were measured. The effects of anthricin on tyrosinase-related protein 1(TYRP1)/TYRP2/tyrosinase (TYR)/microphthalmia-associated transcription factor (MITF) mRNA expressions and melanin content were determined. Quantitative real-time RT-PCR showed that anthricin decreased the mRNA expression level of TYRP1/TYRP2/TYR/MITF genes and melanin production contents than α-MSH-treated B16F10 cells. The tyrosinase activity assay revealed that anthricin decreased the melanin production on the B16F10 cells. These data show that anthricin increases the whitening effects on the B16F10 cells; thus, anthricin is a potent ingredient for skin whitening. Thus, further research on the mechanism of action of anthricin for the development of not only cosmetics, but also healthy food and medicine should be investigated.