• Title/Summary/Keyword: Cell Production

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Growth of Cucumber and Tomato Seedlings by Different Light Intensities and CO2 Concentrations in Closed-type Plant Production System (밀폐형 식물생산시스템 내 CO2와 광도에 따른 오이 및 토마토 묘의 생육)

  • Ji Hye Yun;Hyeon Woo Jeong;So Yeong Hwang;Jin Yu;Hee Sung Hwang;Seung Jae Hwang
    • Journal of Bio-Environment Control
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    • v.32 no.4
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    • pp.257-266
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    • 2023
  • This study was conducted to investigate the growth characteristics of cucumber (Cucumis sativus L. 'Joeunbaekdadagi') and tomato (Solanum lycopersicum L. 'Dotaerang Dia') seedlings by light intensities and CO2 concentrations in a closed-type plant production system (CPPS). Cucumber and tomato seeds were sown in 50-cell trays and germinated in CPPS at air temperature 25 ± 1℃ and relative humidity 50 ± 10% for 4 days. After germination, the CO2 concentrations and light intensity treatment were treated at 500 (ambient), 1,000, and 1,500 µmol·mol-1 and 100, 200, and 300 µmol·m-2·s-1 photosynthetic photon flux density (PPFD), respectively. The leaf area of cucumber showed the highest value in CO2 1,500 μmol·mol-1. However, the leaf area of the tomato had no significant difference in CO2 concentrations and light intensities treatments. In cucumber and tomato both seedlings, the growth and quality such as compactness and leaf area rate were increased with the increase of light intensity, and there were highest in 300 µmol·m-2·s-1. The root surface and number of root tips of cucumber and tomato seedlings were significantly increased with the increase in light intensity. In conclusion, the regulation of the CO2 concentrations and light intensity can control the growth and quality of cucumber and tomato seedlings in CPPS, especially, increasing the light intensity can improve more significantly the growth and quality of seedlings.

Studies on the Post-hatching Development of the Testis in Korean Native Chickens (한국 재래 닭 부화 후 고환 발달에 관한 연구)

  • Jang, B.G.;Tae, H.J.;Choi, C.H.;Park, Y.J.;Park, B.Y.;Park, S.Y.;Kang, H.S.;Kim, N.S.;Lee, Y.H.;Yang, H.H.;Ahn, D.C.;Kim, I.S.
    • Korean Journal of Poultry Science
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    • v.33 no.3
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    • pp.171-179
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    • 2006
  • Changes in the chicken testis from hatching to adulthood were studied in Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The present study was to investigate in more detail the post-hatching development of testis in Korean native chickens. Testes of chickens 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-azure II, qualitative and quantitative(stereological) morphological studies were performed. Sperm production was measured by routine technique. The average volume of a testis of 1 week old Korean native chickens was determined as 0.015 g and the parameter increased linearly from 1 week to 21 weeks days (28.9 g), and did not change from 21 weeks to 64 weeks. The volume density of the seminiferous tubules increased with age from 32.6% at week 1 to 92.89% at week 64. The volume density of the interstitium represents 67.4% of the testicular parenchyma at week 1. This proportion progressively diminished during development to reach a value of 7.11% at week 64. Total sperm production per testis increased significantly from 18 weeks to 28 weeks and remained unchanged. Sperm production per 1 g testis increased significantly from 18 weeks to 28 weeks, did not change significantly from 28 weeks to 52 weeks, and declined significantly at 64 weeks of age. The average diameter of the seminiferous tubules gradually increased with age from 1 week $(42.4{\mu}m)$ to 21 weeks $(412.8{\mu}m)$. The length of the seminiferous tubules was 0.34 m at 1 week, increased significantly in subsequent age groups and reached 72.2 m by weeks 64. The stage of germ cell development in seminiferous tubules was classified as 1) spermatogonia $(1\sim8\;weeks)$, 2) spermatogonia and spermatocytes $(10\sim12\;weeks)$, 3) spermatogonia, spermatocytes and round spermatids $(14\sim16\;weeks)$, and 4) speramatogonia, spermatocytes, spermatids and spermatozoa $(18\sim64\;weeks)$. These results clarified the pattern of changes in the testicular development in Korean native chickens from hatching to adulthood as 1) neonatal-prepubertal $(1\sim12\;weeks)$, 2) puberty$(14\sim18\;weeks)$, and adult$(21\sim64\;weeks)$.

Lipopolysaccharide-induced Synthesis of IL-1beta, IL-6, TNF-alpha and TGF-beta by Peripheral Blood Mononuclear Cells (내독소에 의한 말초혈액 단핵구의 IL-1beta, IL-6, TNF-alpha와 TGF-beta 생성에 관한 연구)

  • Jung, Sung-Hwan;Park, Choon-Sik;Kim, Mi-Ho;Kim, Eun-Young;Chang, Hun-Soo;Ki, Shin-Young;Uh, Soo-Taek;Moon, Seung-Hyuk;Kim, Yang-Hoon;Lee, Hi-Bal
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.846-860
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    • 1998
  • Background: Endotoxin (LPS : lipopolysaccharide), a potent activator of immune system, can induce acute and chronic inflammation through the production of cytokines by a variety of cells, such as monocytes, endothelial cells, lymphocytes, eosinophils, neutrophils and fibroblasts. LPS stimulate the mononucelar cells by two different pathway, the CD14 dependent and independent way, of which the former has been well documented, but not the latter. LPS binds to the LPS-binding protein (LBP), in serum, to make the LPS-LBP complex which interacts with CD14 molecules on the mononuclear cell surface in peripheral blood or is transported to the tissues. In case of high concentration of LPS, LPS can stimulate directly the macrophages without LBP. We investigated to detect the generation of proinflammatory cytokines such as interleukin 1 (IL-1), IL-6 and TNF-$\alpha$ and fibrogenic cytokine, TGF-$\beta$, by peripheral blood mononuclear cells (PBMC) after LPS stimulation under serum-free conditions, which lacks LBPs. Methods : PBMC were obtained by centrifugation on Ficoll Hypaque solution of peripheral venous bloods from healthy normal subjects, then stimulated in the presence of LPS (0.1 ${\mu}g/mL$ to 100 ${\mu}g/mL$ ). The activities of IL-1, IL-6, TNF, and TGF-$\beta$ were measured by bioassaies using cytokines - dependent proliferating or inhibiting cell lines. The cellular sources producing the cytokines was investigated by immunohistochemical stains and in situ hybridization. Results : PBMC started to produce IL-6, TNF-$\alpha$ and TGF-$\beta$ in 1 hr, 4 hrs and 8hrs, respectively, after LPS stimulation. The production of IL-6, TNF-$\alpha$ and TGF-$\beta$ continuously increased 96 hrs after stimulation of LPS. The amount of production was 19.8 ng/ml of IL-6 by $10^5$ PBMC, 4.1 ng/mL of TNF by $10^6$ PBMC and 34.4 pg/mL of TGF-$\beta$ by $2{\times}10^6$ PBMC. The immunoreactivity to IL-6, TNF-$\alpha$ and TGF-$\beta$ were detected on monocytes in LPS-stimulated PBMC. Some of lymphocytes showed positive immunoreactivity to TGF-$\beta$. Double immunohistochemical stain showed that IL-1$\beta$, IL-6, TNF-$\alpha$ expression was not associated with CD14 postivity on monocytes. IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$mRNA expression were same as observed in immunoreactivity for each cytokines. Conclusion: When monocytes are stimulated with LPS under serum-free conditions, IL-6 and TNF-$\alpha$ are secreted in early stage of inflammation. In contrast, the secretion of TGF-$\beta$ arise in the late stages and that is maintained after 96 hrs. The main cells releasing IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$ are monocytes, but also lymphocytes can secret TGF-$\beta$.

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In Vitro Anticancer and Antioxidant Effect of Solvent Extracts from Tuna Dried at Low Temperature Vacuum. (저온진공건조 참치추출물의 in vitro 항암 및 항산화 효과)

  • Jang, Joo-Ri;Kim, Kyung-Kun;Mun, Soo-Beom;Lim, Sun-Young
    • Journal of Life Science
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    • v.19 no.5
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    • pp.633-638
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    • 2009
  • We investigated the inhibitory effects of solvent extracts from dried tuna on the growth of cancer cell lines (HT1080 human fibrosarcoma and HT-29 human colon cancer cells) and $H_2O_2$-induced oxidative stress. Inhibitory effects of acetone with methylene chloride (A+M) and methanol (MeOH) extracts on the growth of HT1080 and HT-29 cancer cells increased in a dose dependent manner (p<0.05). The inhibitory effect was more significant on the growth of HT1080 cells, and A+M extracts had a higher inhibitory effect compared to MeOH extracts. The treatments of hexane, 85% aq. methanol, butanol and water fractions significantly inhibited the growth of both cancer cells (p<0.05). Among the fractions, hexane and 85% aq. methanol fractions showed higher inhibitory effects. In order to determine the protective effect on $H_2O_2$-induced oxidative stress, a DCHF-DA (dichlorodihydrofluorescin diacetate) assay was conducted. All fractions, including crude extracts of dried tuna, appeared to significantly reduce the levels of intracellular reactive oxygen species (ROS) with dose responses (p<0.05). Among the fractions, BuOH and 85% methanol fractions showed a higher protective effect on the production of lipid peroxides. These results indicate that the consumption of tuna may be recommended as a potent functional food for preventing cellular oxidation and cancer.

Effects of Endocrine Disruptors (NP, DBP and BPA) on Sperm Characteristics and Development of IVF Embryos in Pig

  • Yuh, In Suh;Cheong, Hee Tae;Kim, Jong Taek;Park, In Chul;Park, Choon Keun;Yang, Boo Keun
    • Journal of Animal Science and Technology
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    • v.55 no.4
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    • pp.237-247
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    • 2013
  • This study was to examine single or combined in vitro effects of environmental endocrine disruptors on boar sperm characteristics, oxidative stress damage in sperm and development of porcine IVF embryos. Addition of various concentration of NP (10, 20, $30{\mu}M$), DBP (10, 50, $100{\mu}M$) and BPA (1, 5 or $10{\mu}g/ml$) on boar sperm characteristics such as percentages of sperm motility, viability, membrane integrity and mitochondrial activity were dose-dependently decreased within 3, 6 or 9 hr incubation period (p<0.05). The overall detrimental effects increased with incubation time increasement. NP, DBP and BPA showed the detrimental effects on sperm membrane and mitochondria of energy production organelles affecting cell viability with the dependancy of dose and incubation time. In combination effects, NP ($10{\mu}M$) + DBP ($10{\mu}M$) significantly decreased boar general sperm characteristics for 3 or 6 hr incubation period compared with control (p<0.05). When both of NP and DBP concentrations (NP; $30{\mu}M$, DBP; $100{\mu}M$) increase, the detrimental effects on sperm characteristics were larger than those of low concentration combination (p<0.05). The inhibitory effects of NP ($30{\mu}M$) + BPA ($10{\mu}g/ml$) on sperm characteristics were larger than those of NP ($10{\mu}M$) + BPA ($1{\mu}g/ml$) (p<0.05). DBP ($100{\mu}M$) + BPA ($10{\mu}g/ml$) decreased sperm characteristics compared with the low concentration combination (DBP $10{\mu}M$ + BPA $1{\mu}g/ml$, p<0.05). This result indicates the detrimental effects of both chemicals on sperm characteristics were dose dependent. Addition of NP ($30{\mu}M$) + DBP ($100{\mu}M$), NP ($30{\mu}M$) + BPA ($10{\mu}g/ml$), DBP ($10{\mu}M$) + BPA ($1{\mu}g/ml$) or DBP ($100{\mu}M$) + BPA ($10{\mu}g/ml$) significantly increased lipid peroxidation for 3 or 6 hr incubation period (p<0.05) compared with no addition control. NP (${\geq}20{\mu}M$) decreased the percentages of IVF embryo development from morulae and blastocyst stages (p<0.05) and its detrimental effects were dose-dependant. BPA 0, 1, 5 or $10{\mu}g/ml$ decreased significantly and dose-dependently the percentage of morulae plus and blastocysts (p<0.05). Combinations of DBP ($100{\mu}M$) plus NP ($30{\mu}M$) and DBP ($100{\mu}M$) plus BPA ($10{\mu}g/ml$) did not affect on morulae and blastocyst development, but NP ($30{\mu}M$) plus BPA ($10{\mu}g/ml$) has significant detrimental effect on embryo development at these stages (p<0.05). These overall results indicate that the partial detrimental effects on boar sperm characteristics and embryo development by NP, DBP, BPA or the combination of these chemicals might be due to the increasement of lipid peroxidation and free radical formation in the cell and there were no specific interaction effects on boar sperm and embryo degeneration among the combined treatments.

MiR-126-3p inhibits apoptosis and promotes proliferation by targeting phosphatidylinositol 3-kinase regulatory subunit 2 in porcine ovarian granulosa cells

  • Zhou, Xiaofeng;He, Yingting;Jiang, Yao;He, Bo;Deng, Xi;Zhang, Zhe;Yuan, Xiaolong;Li, Jiaqi
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.6
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    • pp.879-887
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    • 2020
  • Objective: Numerous studies have indicated that the apoptosis and proliferation of granulosa cells (GCs) are closely related to the normal growth and development of follicles and ovaries. Previous evidence has suggested that miR-126-3p might get involved in the apoptosis and proliferation of GCs, and phosphatidylinositol 3-kinase regulatory subunit 2 (PIK3R2) gene has been predicted as one target of miR-126-3p. However, the molecular regulation of miR-126-3p on PIK3R2 and the effects of PIK3R2 on porcine GCs apoptosis and proliferation remain virtually unexplored. Methods: In this study, using porcine GCs as a cellular model, luciferase report assay, mutation and deletion were applied to verify the targeting relationship between miR-126-3p and PIK3R2. Annexin-V/PI staining and 5-ethynyl-2'-deoxyuridine assay were applied to explore the effect of PIK3R2 on GCs apoptosis and proliferation, respectively. Real-time quantitative polymerase chain reaction and Western Blot were applied to explore the regulation of miR-126-3p on PIK3R2 expression. Results: We found that miR-126-3p targeted at PIK3R2 and inhibited its mRNA and protein expression. Knockdown of PIK3R2 significantly inhibited the apoptosis and promoted the proliferation of porcine GCs, and significantly down-regulated the mRNA expression of several key genes of PI3K pathway such as insulin-like growth factor 1 receptor (IGF1R), insulin receptor (INSR), pyruvate dehydrogenase kinase 1 (PDK1), and serine/threonine kinase 1 (AKT1). Conclusion: MiR-126-3p might target and inhibit the mRNA and protein expressions of PIK3R2, thereby inhibiting GC apoptosis and promoting GC proliferation by down-regulating several key genes of the PI3K pathway, IGF1R, INSR, PDK1, and AKT1. These findings would provide great insight into further exploring the molecular regulation of miR-126-3p and PIK3R2 on the functions of GCs during the folliculogenesis in female mammals.

Immuno-Enhancing Effect of Enzymatic Extract of Sargassum coreanum Using Crude Enzyme from Shewanella oneidensis PKA 1008 (Shewanella oneidensis PKA 1008 유래 조효소 처리에 의한 큰잎모자반(Sargassum coreanum) 추출 분해물의 면역증진 효과)

  • Park, Sun-Hee;Kim, Min-Ji;Kim, Go-Eun;Park, So-Yeong;Kim, Koth-Bong-Woo-Ri;Kim, Yeon-Ji;Cho, Young-Je;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.8
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    • pp.919-928
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    • 2017
  • The immuno-enhancing effects of alginate oligosaccharides from Sargassum coreanum were investigated. The alginate oligosaccharides were produced by an alginate-degrading enzyme from S. oneidensis PKA 1008. The degraded alginate oligosaccharides were visualized by thin-layer chromatography developed using a solvent system of 1-butanol/methanol/water, 4:1:2 (v/v/v). Alginate was degraded into dimmers at 60 h. As a result, the levels of Th1 cytokine [interferon $(IFN)-{\gamma}$ and interleukin (IL)-2] and Th2 cytokine (IL-6 and IL-10) increased with increasing incubation time compared to the control in vitro. Enzymatic extract treatment promoted proliferation of splenocytes at concentrations of 100 and 200 mg/kg at 24 h in vivo. Secretion of $IFN-{\gamma}$ and IL-2 significantly increased in a dose-dependent manner at 24 h as well as induced higher production of IgG2a in serum. Natural killer cell activity was measured and tended to increase. In addition, complete blood cell counts increased in a dose-dependent manner. These results indicate that alginate oligosaccharides produced by crude enzyme from S. oneidensis PKA 1008 may have significant immune activities.

Physiological Activities of Ginkgo biloba Sarcotesta Extract with Heat Treatment (열처리에 따른 은행 외종피 추출물의 생리활성)

  • Kim, Sung Tae;Lee, Ji Hyun;Lee, Sang Hoon;Jang, Gwi Yeong;Li, Meishan;Kim, Min Young;Yoon, Nara;Lee, Junsoo;Jeong, Heon Sang
    • The Korean Journal of Food And Nutrition
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    • v.28 no.3
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    • pp.369-375
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    • 2015
  • This study was performed to investigate the physiological activities of Ginkgo biloba sarcotesta extracts before and after heat treatment. G. biloba sarcotesta was heated at $130^{\circ}C$ for 2 h and extracted with water, 70% ethanol and 80% methanol. ABTS and DPPH radical scavenging activities increased after heating in the water (14.95 mg AAE/g and 7.36 mg TE/g) and ethanol extracts (12.20 mg AAE/g and 6.23 mg TE/g). ${\alpha}$-Glucosidase inhibitory activity decreased after heating in all but the water extract. Angiotensin converting enzyme I inhibitory activities decreased after heating in all extracts. Nitric oxide production inhibitory activity increased from 12.40~44.55% of the raw sample to 40.76~72.39% of the heated sample at a concentration of $200{\mu}g/mL$. Lipid accumulation inhibitory activities were similar before and after heat treatment. The highest antiproliferative effects on MCF-7 human breast cancer cell lines were observed in 80% methanol extract in the heated sample. Cell viability at concentrations of 25, 50, 100, and $200{\mu}g/mL$ measured 34.88, 17.58, 8.44 and 10.48%, respectively. From the results, the antioxidant and antiproliferative activities of G. biloba sarcotesta extracts increased with heat treatment, and research on the identification of the structure for the active compounds are needed in further studies.

MMP-1 and TIMP-1 production in MG-63 cells stimulated with Prevotella nigrescens Lipopolysaccharide (Prevotella nigrescens lipopolysaccharides로 자극된 MG63 세포에서 분비되는 기질금속단백질 MMP-1과 TIMP-1의 수준에 관한 연구)

  • Yang Won-Kyung;Kim Mi-Ri;Shon Won-Jun;Lee In-Bog;Cho Byeong-Hoon;Um Chung-Moon;Son Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.29 no.5
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    • pp.470-478
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    • 2004
  • The purpose of this study is to monitor the secretion of matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) produced by human osteosarcoma cell line (MG63) stimulated with Prevotella nigrescens lipopolysaccharides (LPS). and to compare the level of secretion before and after the treatment of calcium hydroxide on P. nigrescens LPS. LPS was extracted and purified from anaerobically cultured P. nigrescens. MG63 cells were stimulated by the LPS (0, 1, $10{\;}\mu\textrm{g}/ml$) or LPS($10{\;}\mu\textrm{g}/ml$) pretreated with 12.5 mg/ml of $Ca(OH)_2$ for 3 days. Total RNA was isolated from the cell. and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 and TIMP-1. The results were as follows. 1. MMP-1 mRNA expression at 48 hr was highly increased by stimulation with P. nigrescens LPS. The increase was dose-dependent. 2. When stimulated with ($1{\;}\mu\textrm{g}/ml$ of LPS. TIMP-1 mRNA expression was highly increased at 24 hr and 48 hr. However. TIMP-1 expression was suppressed at higher concentration ($10{\;}\mu\textrm{g}/ml$). 3. When P. nigrescens LPS was pretreated with $Ca(OH)_2$. MMP-1 and TIMP-1 gene expression was downregulated. The results of this study suggest that transcriptional regulation of MMP-1 and TIMP-1 by P. nigrescens LPS could be one of the important mechanisms in bone resorption of periapical inflammation. The result of calcium hydroxide on MMP-1 and TIMP-1 gene expression suppression shows that calcium hydroxide detoxified bacterial LPS and thus should be used the medication of choice for intracanal dressings in root canal infected with black-pigmented bacteria.

Mechanisms of Tributyltin-induced Leydig Cell Apoptosis (유기주석화합물이 웅성생식세포주에 미치는 영향)

  • Lee, Kyung-Jin;Kim, Deok-Song;Ra, Myung-Suk;Wui, Seong-Uk;Im, Wook-Bin;Park, Hueng-Sik;Lee, Jong-Bin
    • Environmental Analysis Health and Toxicology
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    • v.18 no.2
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    • pp.89-94
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    • 2003
  • Tributyltin (TBT) used world-wide in antifouling paints for ships is a widespread environmental pollutant and cause reproductive organs atrophy in rodents. At low doses, antiproliferative modes of action have been shown to be involved, whereas at higher doses apoptosis seems to be the mechanism of toxicity in reproductive organs by TBT. In this study, we investigated that the mechanisms underlying DNA fragmentation induced by TBT in the rat leyding cell line, R2C. Effects of TBT on intracellular Ca$\^$2+/ level and reactive oxygen species (ROS) were investigated in R2C cells by fluorescence detector. TBT significantly induced intracellular Ca$\^$2+/ level in a time-dependent manner. The rise in intracellular Ca$\^$2+/ level was followed by a time-dependent generation of reactive oxygen species (ROS) at the cytosol level. Simultaneously, TBT induced the release of cytochrome c from the mitochondrial membrane into the cytosol. Furthermore, ROS production and the release of cytochrome c were reduced by BAPTA, an intracellular Ca$\^$2+/ chelator, indicating the important role of Ca$\^$2+/ in R2C during these early intracellular events. In addition, Z-DEVD FMK, a caspase-3 inhibitor, decreased apoptosis by TBT. Taken together, the present results indicated that the apoptotic pathway by TBT might start with an increase in intracellular Ca$\^$2+/ level, continues with release of ROS and cytochrome c from mitochondria, activation of caspases,and finally results in DNA fragmentation.