• Title/Summary/Keyword: Serum reduced medium

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Suppressed Cell Proliferation and Differentiation Following an Over-expression of Myostatin is Associated with Inhibited Expression of Insulin-like Growth Factor II and Myogenin in Rat L6 Myoblasts

  • Jin, Eun-Jung;Kim, Inae;Lee, C. Young;Park, Byung-Chul
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.10
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    • pp.1508-1513
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    • 2006
  • Myostatin (MSTN) and insulin-like growth factors (IGFs) are a known inhibitor and stimulators of proliferation and differentiation of muscle cells, respectively. The present study was performed to investigate the relationship of MSTN-induced growth inhibition to expression of the IGF system components and myogenin, a muscle cell-specific transcription factor, in rat L6 myoblasts. The L6 cells transfected with a green fluorescent protein-MSTN plasmid expression construct had a 47% less cell number than mock-transfected cells after 3-d serum-free culture, accompanied by delayed differentiation which was suggested by inhibited aggregation of cells. Moreover, cells transfected with the expression construct had decreased expression of IGF-II and myogenin genes, but not IGF-I or its receptor genes, as examined by reverse transcription-polymerase chain reaction. The reduced mitosis of the L6 cells transfected with the MSTN-expression construct increased following an addition of either IGF-I or IGF-II to the culture medium, but not to the level of mock-transfected cells. By contrast, myogenin gene expression in these cells increased after the addition of either IGF to the level of mock-transfected cells. Collectively, these results suggest that the inhibitory effect of MSTN on L6 cell proliferation and differentiation is likely to be partly mediated by serially suppressed expression of IGF-II and myogenin genes, not IGF-I gene.

Ginseng essence, a medicinal and edible herbal formulation, ameliorates carbon tetrachloride-induced oxidative stress and liver injury in rats

  • Lu, Kuan-Hung;Weng, Ching-Yi;Chen, Wei-Cheng;Sheen, Lee-Yan
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.316-325
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    • 2017
  • Background: Ginseng essence (GE) is a formulation comprising four medicinal and edible herbs including ginseng (Panax ginseng), American ginseng (Panax quinquefolius), lotus seed (Nelumbo nucifera), and lily bulb (Lilium longiflorum). This study was aimed at investigating the hepatoprotective effect of GE against carbon tetrachloride ($CCl_4$)-induced liver injury in rats. Methods: We treated Wistar rats daily with low, medium, and high [0.625 g/kg body weight (bw), 1.25 g/kg bw, and 3.125 g/kg bw, respectively] doses of GE for 9 wk. After the 1st wk of treatment, rats were administered 20% $CCl_4$ (1.5 mL/kg bw) two times a week to induce liver damage until the treatment ended. Results: Serum biochemical analysis indicated that GE ameliorated the elevation of aspartate aminotransferase and alanine aminotransferase and albumin decline in $CCl_4$-treated rats. Moreover, $CCl_4$-induced accumulation of hepatic total cholesterol and triglyceride was inhibited. The hepatoprotective effects of GE involved enhancing the hepatic antioxidant defense system including glutathione, glutathione peroxidase, glutathione reductase, glutathione S-transferase, superoxide dismutase, and catalase. In addition, histological analysis using hematoxylin and eosin and Masson's trichrome staining showed that GE inhibited $CCl_4$-induced hepatic inflammation and fibrosis. Furthermore, immunohistochemical staining of alpha-smooth muscle actin indicated that $CCl_4$-triggered activation of hepatic stellate cells was reduced. Conclusion: These findings demonstrate that GE improves $CCl_4$-induced liver inflammation and fibrosis by attenuating oxidative stress. Therefore, GE could be a promising hepatoprotective herbal formulation for future development of phytotherapy.

Primary Cilium by Polyinosinic:Polycytidylic Acid Regulates the Regenerative Migration of Beas-2B Bronchial Epithelial Cells

  • Gweon, Bomi;Jang, Tae-Kyu;Thuy, Pham Xuan;Moon, Eun-Yi
    • Biomolecules & Therapeutics
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    • v.30 no.2
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    • pp.170-178
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    • 2022
  • The airway epithelium is equipped with the ability to resist respiratory disease development and airway damage, including the migration of airway epithelial cells and the activation of TLR3, which recognizes double-stranded (ds) RNA. Primary cilia on airway epithelial cells are involved in the cell cycle and cell differentiation and repair. In this study, we used Beas-2B human bronchial epithelial cells to investigate the effects of the TLR3 agonist polyinosinic:polycytidylic acid [Poly(I:C)] on airway cell migration and primary cilia (PC) formation. PC formation increased in cells incubated under serum deprivation. Migration was faster in Beas-2B cells pretreated with Poly(I:C) than in control cells, as judged by a wound healing assay, single-cell path tracking, and a Transwell migration assay. No changes in cell migration were observed when the cells were incubated in conditioned medium from Poly(I:C)-treated cells. PC formation was enhanced by Poly(I:C) treatment, but was reduced when the cells were exposed to the ciliogenesis inhibitor ciliobrevin A (CilioA). The inhibition of Beas-2B cell migration by CilioA was also assessed and a slight decrease in ciliogenesis was detected in SARS-CoV-2 spike protein (SP)-treated Beas-2B cells overexpressing ACE2 compared to control cells. Cell migration was decreased by SP but restored by Poly(I:C) treatment. Taken together, our results demonstrate that impaired migration by SP-treated cells can be attenuated by Poly(I:C) treatment, thus increasing airway cell migration through the regulation of ciliogenesis.

The Effects of Alismatis Rhizoma Extract on Allergic Inflammation in RBL-2H3 Mast Cells and OVA/alum-Sensitized Mice (택사 (澤瀉, Alismatis Rhizoma) 추출물이 RBL-2H3 비만세포와 OVA/alum 감작 생쥐의 알레르기 염증 반응에 미치는 영향)

  • Song, Ji Hyun;Lee, Jin Hwa;Kim, Eun Jin;Kim, Yun Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.32 no.3
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    • pp.1-15
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    • 2018
  • Objectives Alismatis Rhizoma has been known to suppress inflammation and allergic reaction. However, the cellular target of Alismatis Rhizoma and its mechanism of action remain unclear. This study was designed to examine the effect of Alismatis Rhizoma extract (ALC) on the RBL-2H3 mast cells in vitro and on the OVA/alum sensitized mice ex vivo. Methods In the study, RBL-2H3 mast cells were cultured in minimal essential medium (MEM) for 24 hours, and treated separately with cyclosporin A and varying doses of ALC, and then stimulated with Phorbol 12-myristate 13-acetate (PMA) (50 ng/ml) and Ionomycin ($0.5{\mu}M$). The levels of IL-13, IL-4 were measured by ELISA analysis. The mRNA levels of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$ were analyzed with Real-time PCR. Also, manifestations of MAPKs transcription factors and $NF-{\kappa}B$ p65 translocation were analyzed by western blotting in vitro. Subsequently, for ex vivo experiment, we induced allergic inflammation on Balb/c mice by OVA/alum and administered ALC orally. And we measured serum OVA-specific IgE level and IL-4, IL-13 in the splenocyte culture supernatant by ELISA analysis. Results ALC was shown to suppress mRNA expression of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$, and to inhibit the IL-13, IL-4 production. Also ALC reduced an activation of mast cells specific signal MAPKs transcription factors and $NF-{\kappa}B$ p65 from the western blot analysis in in vitro experiment. In ex vivo, ALC oral adminstration decreased the level of OVA-specific IgE in serum, and IL-4, IL-13 in the splenocyte culture supernatant. Conclusions ALC is shown to reduce inflammation and allergic response by suppressing Th2 cytokines through the regulation of transcription factors MAPKs and $NF-{\kappa}B$ p65 in mast cells. Administration of ALC suppressed OVA-specific IgE in ovalbumin allergy model through the inhibition of Th2 cytokine. In conclusion, ALC can be considered as an effective treatment for allergic diseases such as atopic dermatitis.

Effect of aging on Leydig cells of Sprague Dawley rats (흰쥐에서 고환 간질세포에 미치는 노화의 영향)

  • Kim, In-Shik;Tae, Hun-Jin;Li, Li-Guang;Park, Young-Jae;Kang, Hyung-Sub;Park, Sang-Youel;Park, Soo-Hyun;Park, Young-Seok;Lee, Young-Hoon;Ahn, Dong-Choon;Choi, Eun-Young;Yang, Hong-Hyun
    • Korean Journal of Veterinary Research
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    • v.43 no.4
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    • pp.541-549
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    • 2003
  • The present study investigated the effects of aging on Leydig cells of Sprague Dawley rats. Rats of 3, 6, 12 and 18 months of age were used. Testes of rat were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in epon-araldite. Using $1{\mu}m$ sections stained with methylene blue, qualitative and quantitative morphological studies were performed. Testis incubations were used to determine luteinizing hormone (LH; 100 ng/ml) stimulated testosterone secretory capacity per testis in vitro. Testosterone levels in the incubation medium, and testosterone and luteinizing hormone levels in serum of these four groups of rats were determined by radioimmunoassay. Morphological studies revealed that Leydig cells were more abundant in the testis interstitium at 6, 12 and 18 months when compared with 3 months. The volumes of Leydig cells per testis was significantly higher, at 6, 12 and 18 months of age than those at 3 months. The number of Leydig cells per testis was doubled at 6, 12 and 18 months of age compared with 3 months. The average volume of a Leydig cell was not significantly different between 3 and 6 months of age, however, at 12 and 18 months a significantly lower value was observed. LH-stimulated testosterone production per testis in vitro was reduced by 45% at 6 months of age compared with 3 months; a further significant reduction was observed at 12 and 18 months. Serum testosterone and LH levels were not significantly different between 3 and 6 months of age but at 12 and 18 months a significantly lower value was observed in both groups for these hormones. These results showed that signs of aging are apparent in Leydig cells of Sprague Dawley rats at 12 months of age.

The Protective Effects of Different Mycotoxin Adsorbents against Blood and Liver Pathological Changes Induced by Mold-contaminated Feed in Broilers

  • Che, Zhengquan;Liu, Yulan;Wang, Huirong;Zhu, Huiling;Hou, Yongqing;Ding, Binying
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.2
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    • pp.250-257
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    • 2011
  • An experiment was conducted to determine the effects of different mycotoxin adsorbents including esterified glucomannan (EGM), hydrated sodium calcium aluminosilicate (HSCAS) and compound mycotoxin adsorbent (CMA) on performance, blood parameters, and liver pathological changes in broilers fed mold-contaminated feed. Two hundred and forty 10-day-old broilers were randomly assigned to one of the five dietary treatments including: i) control diet; ii) mold-contaminated diet; iii) moldcontaminated diet+0.05% EGM; iv) mold-contaminated diet+0.2% HSCAS; v) mold-contaminated diet+0.1% CMA. At 35-days-old, blood and liver tissue samples were collected for analysis. 0.1% CMA improved ADG and ADFI during 10-42 d compared to the moldcontaminated group (p<0.05). The mold-contaminated diet increased total white blood cell (WBC) number, haemoglobin (Hgb) concentration, hematocrit (Hct) level, serum aspartate aminotransferase (AST) and ${\gamma}$-glutamyl transferase (GGT) activities, and decreased red blood cell (RBC) number and serum globulin (GLB) and urea nitrogen (BUN) concentrations (p<0.05). The three mycotoxin adsorbents alleviated the alteration of RBC, WBC, Hgb and AST caused by the mold-contaminated diet. Furthermore, 0.1% CMA increased GLB concentration and decreased Hct level and GGT activity (p<0.05). Liver superoxide dismutase (SOD) activity was reduced, and myeloperoxidase (MPO) activity was increased by the mold-contaminated diet (p<0.05). Both EGM and HSCAS prevented the increase of MPO activity (p<0.05). Liver lesion, including severe vacuolar degeneration of hepatocytes, was observed in chicks fed the mold-contaminated diet. 0.05% EGM prevented these effects except for biliary hyperplasia and mild vacuolar degeneration. 0.2% HSCAS showed medium vacuolar degeneration of hepatocytes. Liver of broilers fed 0.1% CMA revealed a mild vacuolar degeneration. These results indicate that a mold-contaminated diet results in adverse effects on blood parameters and liver morphology. 0.05% EGM and 0.2% HSCAS partially alleviated the adverse effects. However, 0.1% CMA almost completely ameliorated the adverse effects.

Viability of Vibrio parahaemolyticus and V. vulnificus isolated from human in cultured olive flounder (Paralichthys olivaceus) (Vibrio parahaemolyticus와 V. vulnificus 사람 분리균의 넙치, Paralichthys olivaceus에서의 생존율)

  • Im, Su Yeon;Kim, Eunheui
    • Journal of fish pathology
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    • v.35 no.1
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    • pp.57-63
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    • 2022
  • Vibrio parahaemolyticus and V. vulnificus are known to be infected to human via fisheries products. Therefore, food safety of fisheries products is important for public health and fish industry. This paper was conducted to know how well these human isolates can survive in olive flounder (Paralichthys olivaceus). The growth of V. parahaemolyticus and V. vulnificus showed about 50~60% reduced rates at 25℃ than at 37℃ and did not show any differences according to NaCl concentration of media except the increasing in the growth of V. vulnificus in medium containing 3% NaCl. Artificial infection of 1×106 CFU/fish was carried out to confirm the sensitivity of olive flounder against V. parahaemolyticus and V. vulnificus. After 1 week from injection, no fish was dead. To evaluate nonspecific defense of olive flounder against V. parahaemolyticus and V. vulnificus, the antibacterial potency of serum and epidermal mucus were tested. The number of the vibrios exposed to serum obtained from olive flounder significantly decreased after 3 hours, and epidermal mucus showed decrease of the bacteria over than 90% until 12 hours from exposure. Phagocytosis of head kidney leucocytes of healthy olive flounder against V. parahaemolyticus and V. vulnificus showed in over 70% of leucocytes at the 2 hours. Therefore, cultured olive flounder only as vehicle for human pathogen in environmental water is well developed its antibacterial potency against human pathogens, so the viability of V. parahaemolyticus and V. vulnificus in cultured olive flounder was considered very low.

Regulatory Mechanisms of Angiotensin II on the $Na^+/H^+$ Antiport System in Rabbit Renal Proximal Tubule Cells. II. Inhibitory Effects of ANG II on $Na^+$ Uptake

  • Han, Ho-Jae;Park, Soo-Hyun;Koh, Hyun-Ju
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.4
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    • pp.425-434
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    • 1997
  • Many reports represent that angiotensin II (ANG II) caused a dose dependent biphasic effects on fluid transport in the proximal tubule. However, respective roles of different signaling pathways in mediating these effects remain unsettled. The aim of the present study was to examine signaling pathways at high doses of ANG II on the $Na^+$ uptake of primary cultured rabbit renal proximal tubule cells(PTCs) in hormonally defined serum-free medium. High concentrations of ANG II $(>10^{-9}\;M)$ inhibited $Na^+$ uptake and increased $[Ca^{2+}]_i\;level$ in the PTCs. However, low concentrations of $(<10^{-11}\;ANG\;II)$ stimulated $Na^+$ uptake and did not affect $[Ca^{2+}]_i\;level$. 8-(N, N-diethylamino)-octyl-3,3,5- trimethoxybenzoate (TMB-8), ethylene glycol-bis$({/beta}-amino\;ethyl ether)-N,N,N'$, N'-tetra acetic acid (EGTA), and nifedifine partially blocked the inhibitory effects of ANG II on $Na^+$ uptake. When ANG II and bradykinin (BK) were treated together, $Na^+$ uptake was further reduced $(88.47{\pm}1.98%\;of\;that\;of\;ANG\;II,\;81.85{\pm}1.84%\;of\;that\;of\;BK)$. In addition, W-7 and KN-62 blocked the ANG II-induced inhibition of $Na^+$ uptake. Arachidonic acid reduced $Na^+$ uptake in a dose-dependent manner. When ANG II and arachidonic acid were treated together, inhibitory effects on $Na^+$ uptake significantly exhibited greater reduction than that of each group, respectively. When PTCs were treated by mepacrine $(10^{-6}\;M)$ and AACOCF3 $(10^{-5}\;M)$ for 1 hr before the addition of $(<10^{-9}\;ANG\;II)$, the inhibitory effect of ANG II was reversed. In addition, econazole $(>10^{-6}\;M)$ blocked ANG II-induced inhibition of $Na^+$ uptake. In conclusion, the $[Ca^{2+}]_i$ (calcium-calmodulin-dependent kinase) and phospholipase $A_2\;(PLA_2)$ metabolites are involved in the inhibitory effects of ANG II on $Na^+$ uptake in the PTCs.

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Induction of Apoptosis and Growth-Inhibition by Thymoquinone in ACHN and GP-293 Cell Lines in Comparable with Cis-Platinum

  • Shahraki, Samira;Mohebbati, Reza;Shafei, Mohammad Naser;Mahmoudi, Mahmoud;Hosseinian, Sara;Parhizgar, Soghra;Yazd, Zohreh Naji Ebrahimi;Heravi, Nazanin Entezari;Abadi, Reza Nejad Shahrokh;Khajavirad, Abolfazl
    • Journal of Pharmacopuncture
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    • v.22 no.3
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    • pp.176-183
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    • 2019
  • Objective: In the current work, we investigated the cytotoxic and apoptotic effects of Thymoquinone (TQ), an active compound of Nigella sativa (N. sativa) and Cis-platinum, on normal renal epithelial (GP-293) and human renal adenocarcinoma cell lines (ACHN). Methods: GP-293 and ACHN cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% Fetal bovine serum (FBS) and 1% penicillin plus streptomycin antibiotic. The MTT assay was used for cellular viability assessment. Viability of cells was observed using inverted light microscope 24, 48 and 72 h after exposure of the cells to various concentrations of TQ (1, 2.5, 5, 10, 50 and $100{\mu}g/ml$) and Cis-platinum (0.5, 1, 1.5, 2, 3, 6 and $12.5{\mu}g/ml$). Moreover, apoptosis was analyzed with a flow-cytometry method. The untreated cells were considered as control group. Results: Morphological changes such as reduced cell number and increased intercellular distance and reduced cell viability in ACHN and GP-293cell lines were observed in both TQ and Cis- platinum groups; however, Cis-platinum had greater effect on ACHN cell line than GP-293 cell line. In addition, GP-293 cell line was more sensitive to TQ compared to ACHN cell line. Furthermore, TQ and Cis-platinum had apoptotic effects on both ACHN and GP-293 cell lines. Conclusion: Our findings demonstrated that TQ and Cis-platinum had cytotoxic and apoptotic effects on both cell lines, However, GP-293 cell line was more sensitive to TQ. Additionally, Cis-platinum was more effective on ACHN cell line than on GP-293 cell line.

A Concentrated Onion Extract Lowers Serum Lipid Levels in Rats Fed a High-Fat Diet (흰쥐에서 양파 농축액의 고지혈 개선 작용)

  • Kim, Ju-Youn;Seo, Yun-Jung;Noh, Sang-K.;Cha, Yong-Jun
    • Food Science and Preservation
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    • v.17 no.3
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    • pp.398-404
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    • 2010
  • It is known that onions, or bioactive compounds therein, providehealth benefits. The present study was designed to investigate whether a concentrated onion extract lowered blood lipid levels in rats fed a high-fat diet. Initially, male Sprague-Dawley rats were housed singly in an environment in which temperature and light duration were controlled, and had free access to a nutritionally adequate AIN-93G diet and deionized water. After an acclimatization period, rats were weight-matched and assigned to one of the following five groups: 1) a control group, fed the AIN-93G diet mixed with 10% (w/v) lard and 0.7% (w/v) cholesterol to induce hyperlipidemia (control); 2) three experimental groups, fed the AIN-93G diet mixed with a high-fat source plus concentrated onion extract at three different levels (termed the low-dose, medium-dose, and high-dose groups); and, 3) a placebo group, fed the AIN-93G diet with fats plus the same concentrated extract but devoid of onion-derived material. All five groups freely ingested their respective diets over 6 weeks. At 0, 3, and 6 weeks, blood samples were collected from the orbital sinus following overnight food deprivation. At 6 weeks, livers were collected. Both control and experimental groups continually gained body weight throughout the study. No significant differencein body weight gain was observed among groups. However, the serum concentrations of triglycerides, total cholesterol, and non HDL-cholesterol were significantly reduced by ingestion of concentrated onion extract. Also, the hepatic levels of total lipids and total fatty acids, especially C18:1 (oleic acid), were significantly decreased in rats fed a high level of concentrated onion extract, compared with the control and placebo groups. These results provide clear evidence that ingestion of a concentrated onion extract has a profound inhibitory effect on serum lipid levels in rats fed a high-fat diet. Our findings indicate that a concentrated onion extract may be used to alleviate hyperlipidemia by lowering serum cholesterol and triglyceride levels.