• Title/Summary/Keyword: apoptotic cells

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Effects of High-fat Diet on Type-I Muscle Loss in Rats (고지방식이가 쥐의 Type-I 근육손실에 미치는 영향)

  • Baek, Kyung-Wan;Cha, Hee-Jae;Park, Jung-Jun
    • Journal of Life Science
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    • v.23 no.12
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    • pp.1509-1515
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    • 2013
  • The term lipotoxicity has been used to describe how excess lipid accumulation leads to cellular dysfunction and death in non-adipose tissues, including skeletal muscle. While lipotoxicity has been found in cultured skeletal muscle cells with high-fat feeding, the consequences of lipotoxicity in vivo are still unknown, particularly in Type-I muscle, which is metabolically affected by lipotoxicity. The aim of this study was to investigate the effects of a high-fat diet on changes in the morphology and apoptotic protein expression of Type-I muscle loss in rats. The rats were fed either a high-fat diet or a normal diet for six weeks, and then lipid accumulation, inflammation response, and nucleus infiltration were measured, and PARP protein expression was cleaved by Oil Red O staining, H & E staining, and Western blot, respectively. Lipid accumulation, inflammation response, nucleus infiltration, and cleaved PARP protein expression were significantly (p<0.05) higher in the high-fat diet group than they were in the normal diet group. The weight of Type-I muscle tended to be lower in the high-fat diet group compared to the normal diet group, but the difference was not statistically significant. These results indicate that a high-fat diet triggers cell death in Type-I muscle via lipotoxicity, which suggests that a high-fat diet may be associated with sarcopenia.

Effect of Haedokjeongki-tang in Rat Liver after Monocrotaline Exposure (Monocrotaline으로 유발된 흰쥐의 간독성에 대한 해독정기탕의 효과)

  • Park, Hyun-Jeong;Kim, Jeong-Sang
    • Applied Microscopy
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    • v.37 no.1
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    • pp.1-10
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    • 2007
  • Monocrotaline (MCT) is a pyrrolizidine alkaloid (PA) plant toxin that produces hapatotoxicity in humans and animals. To felt the hypothesis, we investigated the possible protective effects of Haedokjeongki-tang as an antioxidant against MCT-induced liver injury in rats. Cells with apoptotic morphology have been observed in the livers of animals exposed to Ph and Haedokjeongki-tang. Whether apoptosis occurs in the livers of MCT-treated animals and whether it is required for full manifestation of pathological changes is not known, To determine this, rats were treated with 100 mg MCT/kg, and apoptosis was detected by transmission electron microscopy and TUNEL assay. MCT produced apoptosis in the liver by 6 h after treatment and increased by 24 h. Administration of Haedokjeongki-tang did affect liver structure and inhibit apopotosis in MCT-induced liver injury. Upon light and electron microscopic examination, we observed that Haedokjeongki-tang improved the morphological and histopathological changes of the liver caused by MCT-induced injury. MCT caused a time-dependent release of GOT and GPT, a marker of liver injury. Furthermore, we observed with respect to antioxidants status, catalase and superoxide dismutase activity tended to be higher in the MCT-treated rats than in the Haedokjeongki-tang administered rats. Our finding showed that Haedokjeongki-tang administration partially reduced liver injury after MCT exposure.

Effect of L-Glutathione Treatment during Somatic Cell Nuclear Transfer Procedures on the Subsequent Embryonic Development and DNA Methylation Status of Cloned Bovine Embryos

  • Bae, Hyo-Kyung;Yoon, Nam-Sik;Hwang, In-Sun;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Journal of Embryo Transfer
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    • v.29 no.4
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    • pp.345-350
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    • 2014
  • We investigate the effect of L-glutathione (GSH), an antioxidant, treatment during the somatic cell nuclear transfer (SCNT) procedures on the in vitro development and DNA methylation status of bovine SCNT embryos. Bovine in vitro matured (IVM) oocytes were enucleated and electrofused with a donor cell, then activated by a combination of Ca-ionophore and 6-dimethylaminopurine. The recipient oocytes or reconstituted oocytes were treated with $50{\mu}M$ GSH during these SCNT procedures from enucleation to activation treatment. The SCNT embryos were cultured for 7 days to evaluate the in vitro development, apoptosis and DNA methylation in blastocysts. The apoptosis was measured by TUNEL assay and caspase-3 activity assay. Methylated DNA of SCNT embryos at the blastocyst stages was detected using a 5-methylcytidine (5-MeC) antibody. The developmental rate to the blastocyst stage was significantly higher (P<0.05) in GSH treatment group ($32.5{\pm}1.2%$, 78/235) than that of non-treated control SCNT embryos ($22.3{\pm}1.8%$, 50/224). TUNEL assay revealed that the numbers of apoptotic cells in GSH treatment group ($2.3{\pm}0.4%$) were significantly lower (P<0.05) than that of control ($3.8{\pm}0.6%$). Relative caspase-3 activity of GSH treated group was $0.8{\pm}0.06$ fold compared to that of control. DNA methylation status of blastocysts in GSH treatment group ($13.1{\pm}0.5$, pixels/embryo) was significantly lower (P<0.05) than that of control ($17.4{\pm}0.9$, pixels/embryo). These results suggest that antioxidant GSH treatment during SCNT procedures can improve the embryonic development and reduce the apoptosis and DNA methylation level of bovine SCNT embryos, which may enhance the nuclear reprogramming of bovine SCNT embryos.

Effects of Heat Stress on the Developmental Competence of Bovine Cumulus-Oocyte Complex During in vitro Maturation (Heat Stress가 소 난자의 체외성숙과 배반포 발달에 미치는 영향)

  • Kim, Min-Su;Kim, Chan-Lan;Seong, Hwan-Hoo;Kim, Namtae;Kim, Sung Woo
    • Journal of Embryo Transfer
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    • v.32 no.3
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    • pp.65-71
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    • 2017
  • The elevated temperature and high humidity has been known as main reason for heat stress on animals and cause detrimental effects on productivity of organisms and physiological conditions of normal bioactivities. The aims of this study were to evaluate the relationship between time of heat shock simulation during in vitro maturation and developmental competence of subsequent embryo after in vitro fertilization. Heat shocked cumulus-oocyte complexes (COCs) of Korean native cattle were subjected to normal conditions for 22, 21, 18 and 12 h respectively and transferred to heat stress inducing condition at $40.5^{\circ}C$ in other incubator for 0 (control), 1 and 4 h. After maturation for 22 h, the oocytes were fertilized and cultured in mSOF media for 8 d and examined the developmental capacity of embryos. There were no differences in maturation and cleavage rates between 0, 1 and 4 h heat socked oocytes, but blastocysts formation were lower in the 4 h heat stressed oocytes. The apoptotic cells of developed blastocysts were also increased in at day 8 with 4 h heat shocked oocytes. These results indicate that heat shock on oocytes during maturation could cause negative effects on the developmental competence of embryos.

Drying seaweeds using hybrid hot water Goodle dryer (HHGD): comparison with freeze-dryer in chemical composition and antioxidant activity

  • Nagahawatta, D.P.;Asanka Sanjeewa, K.K.;Jayawardena, Thilina U.;Kim, Hyun-Soo;Yang, Hye-Won;Jiang, Yunfei;Je, Jun-Geon;Lee, Tae-Ki;Jeon, You-Jin
    • Fisheries and Aquatic Sciences
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    • v.24 no.1
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    • pp.19-31
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    • 2021
  • Seaweeds are a potential source of minerals, essential amino acids, fatty acids, proteins, and various bioactive compounds such as antioxidants. The higher water content of seaweeds reduces the shelf life and this requires the appropriate drying method. The drying conditions play a major role in the conservation of nutrient composition in dried seaweeds. In recent years, the seaweed industry has used many different drying methods with advantages and limitations. Hybrid hot-water Goodle dryer (HHGD) which is a special dryer mixed with hot-water and a Korean traditional heating system (Goodlejang) might be a solution to avoid these limitations. The present study evaluated the effect of drying conditions in HHGD on nutrient composition and bioactivities of brown seaweeds. Moreover, freeze-dryer (FD) and HHGD were employed in this study to compare the dried outputs obtained from four brown seaweed species. The present study aims to evaluate the effect of the hybrid hot-water Goodle drying method (HHGDM) on the nutritional composition and antioxidant activity of dried seaweeds. AOAC standard methods were used to analyze the proximate composition of dried samples and their 70% ethanol extract. The intracellular and extracellular antioxidant activities were evaluated using Vero cells and electron spin resonance (ESR) spectrometer respectively. High performance liquid chromatography, apoptotic body formation, and in-vivo experiments were used for further confirmation of the quality of dried output. The proximate composition results obtained from drying in HHGD and FD did not exhibit any significant difference. Moreover, the seaweed extracts from the dried seaweeds by HHGD and FD dryings were also not different and both significantly down-regulated in-vivo and in-vitro oxidative stress. Furthermore, the high performance liquid chromatography results revealed that the two dryers did not make the major peaks different in the chromatograms. Freeze-drying method (FDM) provides elevated quality for dried output, but there are limitations such as high cost and low capacity. The results from a novel HHGD did not provide any significant difference with the results in FD and expressed a potential to avoid the limitations in FD. Overall, these findings solidified the applicability of HHGD over FD.

Protodioscin protects porcine oocytes against H2O2-induced oxidative stress during in vitro maturation

  • So-Hee Kim;Seung-Eun Lee;Jae-Wook Yoon;Hyo-Jin Park;Seung-Hwan Oh;Do-Geon Lee;Da-Bin Pyeon;Eun-Young Kim;Se-Pill Park
    • Animal Bioscience
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    • v.36 no.5
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    • pp.710-719
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    • 2023
  • Objective: The present study investigated whether protodioscin (PD), a steroidal saponin mainly found in rhizome of Dioscorea species, alleviates oxidative stress-induced damage of porcine oocytes during in vitro maturation. Methods: Oocytes were treated with different concentrations of PD (0, 1, 10, 100, and 200 µM) in the presence of 200 µM H2O2 during in vitro maturation. Following maturation, spindle morphology and mitogen-activated protein kinase activity was assessed along with reactive oxygen species level, GSH activity, and mRNA expression of endogenous antioxidant genes at the MII stage. On the day 7 after parthenogenetic activation, blastocyst formation rate was calculated and the quality of embryo and mRNA expression of development-related genes was evaluated. Results: Developmental competence was significantly poorer in the 0 µM PD-treated (control) group than in the non-treated (normal) and 10 µM PD-treated (10PD) groups. Although the reactive oxygen species level did not significantly differ between these three groups, the glutathione level and mRNA expression of antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, nuclear factor erythroid 2-related factor 2 [Nrf2], and hemo oxygenase-1 [HO-1]) were significantly higher in the normal and 10PD groups than in the control group. In addition, the percentage of oocytes with defective spindle and abnormal chromosomal alignment was significantly lower and the ratio of phosphorylated p44/42 to total p44/42 was significantly higher in the normal and 10PD groups than in the control group. The total cell number per blastocyst was significantly higher in the 10PD group than in the control group. The percentage of apoptotic cells in blastocysts was highest in the control group; however, the difference was not significant. mRNA expression of development-related genes (POU domain, class 5, transcription factor 1 [POU5F1], caudal type homeobox 2 [CDX2], Nanog homeobox [NANOG]) was consistently increased by addition of PD. Conclusion: The PD effectively improves the developmental competence and quality of blastocysts by protecting porcine oocytes against oxidative stress.

Synergism Induced by Combination of Farnesyl Transferase Inhibitor SCH66336 and Insulin like-Growth Factor Binding Protein-3 in apoptosis of Non-Small Cell Lung Cancer Cell lines (비소세포성 폐암 세포주에서 Farnesyl Transferase Inhibitor SCH66336과 인슐린양 성장 인자 결합 단백-3의 병용처리에 의한 세포고사 상승 작용)

  • Kim, Young;Kim, Se Kyu;Kim, Hyung Jung;Chang, Joon;Ahn, Chul Min;Kim, Sung Kyu;Chang, Yoon Soo
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.2
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    • pp.120-128
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    • 2005
  • Background : Insulin-like growth factor binding protein (IGFBP)-3 regulates non-small cell lung cancer(NSCLC) cell proliferation in vitro and in vivo by inhibiting IGF-mediated signaling pathways. To have better strategies for the treatment of lung cancer, we analyzed the combining effects of adenovirus expressing IGFBP-3 (Ad5CMV-BP3) and SCH66336, a farnesyl transferase inhibitor (FTI) designed to block Ras-mediated proliferative signaling pathways. Methods : To measure the combining effects of Ad5CMV-BP3 and SCH66336 on the proliferation of NSCLC cells, human NSCLC cell lines (H1299, H596, A549, H460, and H358), SCH66336, recombinant adenovirus expressing IGFBP-3 (Ad5CMV-BP3) and athymic nude mice were used in these experiments. Results : The combination of Ad5CMV-BP3 and SCH66336 produced a synergistic enhancement in antiproliferative effects over a range of clinically achievable concentrations in a variety of NSCLC cell lines. Furthermore, we observed a significant reduction in growth of NSCLC xenograft induced in athymic nude mice. Conclusion : In conclusion, this study demonstrated for the first time that the FTI SCH66336 synergizes with IGFBP-3 and enhances its apoptotic activity in NSCLC cells in vitro and in vivo. The combined treatment of Ad5CMV-BP3 and SCH66336 raises the possibility of using this regimen in clinic for the treatment of NSCLC.

Cytotoxicity of COX-2 Inhibitor (Nimesulide) in Non-small Cell Lung Cancer Cell Line (비소세포폐암 세포주에서 COX-2억제제(Nimesulide)의 세포독성)

  • Park Chan Beom;Jeon Hyun Woo;Jin Ung;Cho Kyu Do;Kim Chi Kyung;Wang Young-Pil
    • Journal of Chest Surgery
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    • v.38 no.4 s.249
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    • pp.263-270
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    • 2005
  • In recent years, a combination of two demographic phenomena, an increased number of older people in the population and an increase in the incidence of lung cancer with age, has made it mandatory to develop therapeutic modalities with less toxicity for the treatment of inoperable elderly patients with lung cancer. Therefore, we investigated the correlation between COX-2 expression and cytotoxicity of Nimesulide, a specific COX-2 inhibitor. Material and Method: Immunohistochemical staining of COX-2 was performed. After exposure of Nimesulide, XTT analysis, FACS analysis and Hoechst staining were carried out. Result: COX-2 protein was expressed in non-treated A549 cells strongly, but not in H1299. Cytotoxicity of Nimesulide against A549 cell and H1299 cell were similar and $IC_{50}$ of Nimesulide in both cell lines were $70.9{\mu}M$ in A549 cell line and $56.5{\mu}M$ in H1299 cell line respectively. FACS analysis showed $G_0/G_1$ arrest in both cell lines and the S phase cell fraction was decreased. Morphologic assessment of apoptosis by Hoechst 33258 staining, many apoptotic cells were detected in both cell lines. Conclusion: Selective COX-2 inhibitor, Nimesulide, can inhibit the proliferation of non-small cell lung cancer cell lines in vitro. Inhibitory effect of Nimesulide are induction of apoptosis and $G_0/G_1$ arrest. There is no correlation between COX-2 expression and cytotoxicity of Nimesulide, a specific COX-2 inhibitor. Therefore, highly selective COX-2 inhibitors such as Nimesulide can be expected to lead to even greater efficacy of their use as adjuncts to various anticancer angents and radiation therapy for the treatment of high-risk patients.

Taurine exerts neuroprotective effects via anti-apoptosis in hypoxic-ischemic brain injury in neonatal rats (신생 흰쥐의 저산소성 허혈성 뇌손상에서 항세포사멸사를 통한 taurine의 신경보호 효과)

  • Jeong, Ji Eun;Kim, Tae Yeol;Park, Hye Jin;Lee, Kye Hyang;Lee, Kyung Hoon;Choi, Eun Jin;Kim, Jin Kyung;Chung, Hai Lee;Seo, Eok Su;Kim, Woo Taek
    • Clinical and Experimental Pediatrics
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    • v.52 no.12
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    • pp.1337-1347
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    • 2009
  • Purpose:Taurine (2-aminoethanesulfonic acid) is a simple sulfur-containing amino acid. It is abundantly present in tissues such as brain, retina, heart, and skeletal muscles. Current studies have demonstrated the neuroprotective effects of taurine, but limited data are available for such effects during neonatal period. The aim of this study was to determine whether taurine could reduce hypoxic-ischemic (HI) cerebral injury via anti-apoptosis mechanism. Methods:Embryonic cortical neurons isolated from Sprague-Dawley (SD) rats at 18 days gestation were cultured in vitro. The cells were divided into hypoxia group, taurine-treated group before hypoxic insult, and taurine-treated group after HI insult. In the in vivo model, left carotid artery ligation was performed in 7-day-old SD rat pups. The pups were exposed to hypoxia, administered an injection of 30 mg/kg of taurine, and killed at 1 day, 3 days, 1 week, 2 weeks, and 4 weeks after the hypoxic insult. We compared the expressions of Bcl-2, Bax, and caspase-3 among the 3 groups by using real- time polymerase chain reaction (PCR) and western blotting. Results:The cells in the taurine-treated group before hypoxic insult, although similar in appearance to those in the normoxia group, were lesser in number. In the taurine-treated group, Bcl-2 expression increased, whereas Bax and caspase-3 expressions reduced. Conclusion:Taurine exerts neuroprotective effects onperinatal HI brain injury due to its anti-apoptotic effect. The neuroprotective effect was maximal at 1-2 weeks after the hypoxic injury.

Cell Protective Effects of Enzymatic Hydrolysates of Citrus Peel Pectin (귤피 펙틴 유래 효소적 가수분해물의 세포 보호 효과)

  • Kwon, Soon Woo;Ko, Hyun Ju;Bae, Jun Tae;Kim, Jin Hwa;Lee, Geun Soo;Pyo, Hyeong Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.1
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    • pp.75-85
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    • 2016
  • Pectin, a naturally occurring polysaccharide, has in recent years attracted considerable attention. Its benefits are increasingly appreciated by scientists and consumers due to its safety and usefulness. The chemistry and gel-forming characteristics of pectin have enabled to be used in pharmaceutical industry, health promotion and treatment. Yet, it has been rarely used in cosmetics because of its incompatibility with many cosmetic ingredients, including alcohols, and unstable viscosity of pectin gels under various pH and salt conditions. However, low-molecular-weight pectin oligomers have excellent biological activities, and depolymerization of pectin to produce cosmetic ingredients would be very useful. In this study, we attempted the development of cosmetic ingredients using pectin with an excellent effect on human skin. We developed a bio-conversion process that uses enzymatic hydrolysis to produce pectin hydrolysates containing mainly low-molecular-weight pectin oligomers. Gel permeation chromatography was used to determined the ratio of hydrolysis. The molecular weight of the pectin hydrolysates obtained varied between 200 and 2,700 Da. The two newly developed low-molecular-weight pectin hydrolysates, LMPH A and B, had higher anti-oxidative activities than pectin or D-galacturonic. Exposure to UVB radiation induces apoptotic cell death in epidermal cells. Annexin V binding and propidium iodide uptake were measured by flow cytometry to evaluate UVB-induced cell death in HaCaT cells. Both LMPH A and B reduced UVB-induced cell death and increased cell proliferation by 22% and 30% at 0.5% concentration respectively, while pectin had no significant activity. In conclusion, this study suggests that the newly developed low-molecular-weight pectin hydrolysates can be used as safe and biologically active cosmetic ingredients.