• Title/Summary/Keyword: Tunel assay

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Protective Effects of Singihwan (腎氣丸) on Traumatic Brain Injury-induced Apoptosis in Rat Hippocampal Dentate Gyrus

  • Kwon, Oh-Bong;Song, Yun-kyung;Lim, Hyung-Ho
    • The Journal of Korean Medicine
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    • v.29 no.2
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    • pp.21-31
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    • 2008
  • Backgrounds: Singihwan is used "to strengthen inborn energy" and we suspected a protective effect on brain neuron cells. Objectives: The aim of this study was to evaluate the effects of Singihwan (SGH) on traumatic brain injury-induced delayed apoptosis in rat hippocampal dentate gyrus. Methods: For a surgical induction of traumatic brain injury (TBI), a 5 mm diameter stainless rod was used to make traumatic attack from the surface of the brain used by an impactor. The protective effect of the aqueous extract of SGH against TBI in the rat hippocampal dentate gyrus was investigated by using step-down avoidance task, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, Bax immunohistochemistry, and 5-bromo-2'-deoxyuridine (BrdU) immunohistochemistry. Results: The aqueous extract of SGH suppressed the TBI-induced increase in apoptosis and cell proliferation in the hippocampal dentate gyrus. Conclusions: It is possible that the aqueous extract of SGH has a neuroprotective effect on TBI-induced neuronal cell death.

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Adenovirus-Mediated Antisense Expression of Telomerase Template RNA Induces Apoptosis in Lung Cancer Cells

  • Song, Joon-Seok;Kim, Sang-Bae;Lee, Young-Ho;Lee, Kyu-Wan;Jung, Hak-Huyn;Kim, Mee-Hye;Kim, Kyung-Tai;Brown, Robert;Kim, Young-Tae
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.89-95
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    • 2002
  • Telomerase is a ribonucleoprotein complex, whose function is to add telomeric repeats $(TTAGGG)_n$ to chromosomal ends and is also known to play an important role in cellular immortalization. Telomerase is highly active in most tumor cells, yet not in normal cells. Therefore, it may have possible applications in cancer gene therapy. Telomerase consists of two essential components; a telomerase RNA template (hTR) and a catalytic subunit (hTERT). The current study attempted to inhibit the "open" part of the human telomerase RNA (hTR) with an antisense sequence-expressing adenovirus. It was found that the antisense telomerase adenovirus suppressed the telomerase activity, tumor cell growth, and survival in vitro. Furthermore, FACS analysis and TUNEL assay suggested that the reduce viability was mediated through the induction of apoptosis, indicating that this approach might be a useful method for suppressing cancer growth in targeted cancer gene therapy.

Study on Changes in Endogenous Stem Cells in the Salivary Gland of Streptozotocin-induced Diabetic Rats

  • Jung, Bo Hyun;Lee, Hee Su;Yoo, Ki-Yeon
    • International Journal of Oral Biology
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    • v.42 no.3
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    • pp.99-106
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    • 2017
  • Type1 diabetes mellitus (DM) is generally known to be caused by destruction of insulin-producing pancreatic ${\beta}$ cells or an immune-related problem. Polydipsia is a representative symptom of DM, and it has been reported that this condition is closely related to xerostomia and is considered that hyposalivation from the salivary gland results in this phenomenon. Although various studies have reported that induction of diabetes reduces endogenous stem cells in other organs (heart, brain etc.), diabetes-related changes in endogenous stem cells in the salivary gland have not yet been well established. Therefore, in this study, to verify the change in salivary gland stem cells after diabetes, salivary gland tissues in the control and diabetes-induced groups were processed by histochemistry (Masson's trichrome staining) for morphological analysis, TUNEL assay for cell death, and immunohistochemistry (Ki-67 and c-Kit) for cell proliferation and maturation. Diabetes induced by STZ leads to vacuolization, apoptosis, and reduction in proliferating cells/salivary gland stem cells in salivary glands of rats. This result suggests that diabetes may be associated with reduction in salivary gland function such as degeneration and inhibition of regeneration in the salivary gland.

Comparison of the Effects of 13-cis Retinoic Acid and Melatonin on the Viabilities of SH-SY5Y Neuroblastoma Cell Line

  • Tosun, Murat;Soysal, Yasemin;Mas, Nuket Gocmen;Karabekir, Hamit Selim
    • Journal of Korean Neurosurgical Society
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    • v.57 no.3
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    • pp.147-151
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    • 2015
  • Objective : Neuroblastoma is one of common childhood tumors. Although its mortality is very high, there is no effective treatment yet. The aim of this project is to evaluate cytotoxic effects of melatonin (MLT) an endogen hormone and 13-cis retinoic acid (13-cis-RA) also named as isotretinoin an analogue of vitamin A on neuroblastoma SH-SY5Y cell line. Methods : In this study, SH-SY5Y cell line was used. After cell culture, the cells were exposed to different doses of MLT and 13-cis-RA. 24 and 48 hours later. While the viabilities was estimated with MTT cell viability assay test, apoptotic indexes were calculated after staining with TUNEL based apoptosis kit. Results : It was observed that MLT has very effective cytotoxic potential than 13-cis-RA on neuroblastoma cell line. At the same time, when MLT and 13-cis-RA were combined, this effect was potentiated. On the other hand, it was found that the effect of 13-cis-RA individually on neuroblastoma cells was very slight. Conclusion : We suggest that in the treatment of patient with neuroblastoma, MLT is very effective and also this effect can be augmented by combination with 13-cis-RA.

Anti-apoptosis effects by Eimeria tenella infection in Madin-Darby bovine kidney cells

  • Lee, Hyun-A;Hong, Sun-Hwa;Chung, Yung-Ho;Kim, Ok-Jin
    • Korean Journal of Veterinary Service
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    • v.35 no.2
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    • pp.105-109
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    • 2012
  • Apoptosis is a host defense mechanism that the cell uses to limit production of infectious pathogens. Although many bacteria, viruses and parasites can induce apoptosis in infected cells, some pathogens usually exhibit the ability to suppress the induction of apoptosis in the infected cells. Sophisticated evasion strategies of obligate intracellular parasites, in particular prevention of host cell apoptosis, are necessary to ensure successful replication. To study the ability of Eimeria tenella in this regard, in vitro experiments were performed applying Madin-Darby bovine kidney (MDBK) cells as host cell. We have demonstrated that productive infection of adherent cell lines by E. tenella resulted in an anti-apototic effect. This phenomenon was confirmed using in situ terminal deoxynucleotidyl transferase-mediated (TdT) deoxyuridine triphosphates (dUTP)-fluorescein nick end labeling (TUNEL) assay to detect apoptosis. Therefore, E. tenella could complete its cycle of productive infection while inducing anti-apoptosis in the infected cells. This finding might have implications for the pathobiology of E. tenella and other Eimeria species.

The mechanism of apoptosis induced by eugenol in human osteosarcoma cells

  • Shin, Sang-Hun;Park, Jae-Hyun;Kim, Gyoo-Cheon;Park, Bong-Soo;Gil, Young-Gi;Kim, Chul-Hoon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.1
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    • pp.20-27
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    • 2007
  • Eugenol is commonly used in dentistry for the sedation of toothache, pulpitis, and dental hyperalgesia. This study was performed to investigate the apoptotic effect of eugenol to human osteosarcoma (HOS) cells and the potential use of this compound in osteosarcoma cells. Eugenol showed the apoptotic effect in HOS cells in dose- and time-dependent manner. Fragmentation and condensation of DNA were showed by TUNEL assay, Hemacolor stain and Hoechst stain. In the DNA electrophoresis analysis, cells showed DNA degradation characteristic of apoptosis with a ladder pattern of DNA fragments. Apoptosis-related factors were analyzed by western blotting. Cells treated with eugenol showed caspase-3, PARP, lamin A and DFF-45 cleavage. Eugenol treatment induced caspase-3 cleavage and activation. Cleavages of PARP, DFF-45 and lamin A were accompanied with activation of caspase triggered by eugenol in HOS cells. Though this study needs more investigations, these results suggest that eugenol induce apoptosis via caspase dependent pathway in HOS cells and eugenol may constitute a potential antitumor compound against osteosarcoma cells.

The Protective Effects of N-Acetyl-L-cysteine on Cadmium-induced Cell Apoptosis in Rat Testis

  • Kim, Ji-Sun;Soh, Jaemog
    • Biomedical Science Letters
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    • v.25 no.4
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    • pp.417-425
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    • 2019
  • Cadmium (Cd) generates reactive oxygen species (ROS), which in turn cause the apoptosis of various cell types including developing germ cells in rodent testis. Ascorbic acids (AA), one of the ROS scavengers, had been reported to protect against Cd-induced apoptosis. N-Acetyl-L-cysteine (NAC), another ROS scavenger, is known to remove ROS and alleviate the Cd-induced apoptosis in various cell types. In this study we tried to elucidate how NAC affected on Cd-induced cell apoptosis in rat testis. Rats were administered with NAC before and after Cd treatment and then testicular cell apoptosis was examined. NAC treatment resulted in the reduction of Cd-induced chromosomal DNA fragmentation in agarose gel electrophoresis. Terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) assay showed that treatment of NAC reduced the Cd-induced apoptosis of germ cells. The administration of NAC showed that the translocation of apoptosis inducing factor (AIF) from mitochondria to nucleus was prevented, which indicated that the mechanism of Cd-induced testicular apoptosis is mediated through the release of AIF in caspase-independent manner. Taken together, the NAC may remove Cd-induced ROS and protect ROS-induced cell apoptosis in rat testis.

Abrin Induces HeLa Cell Apoptosis by Cytochrome c Release and Caspase Activation

  • Qu, Xiaoling;Qing, Liuting
    • BMB Reports
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    • v.37 no.4
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    • pp.445-453
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    • 2004
  • We identified apoptosis as being a significant mechanism of toxicity following the exposure of HeLa cell cultures to abrin holotoxin, which is in addition to its inhibition of protein biosynthesis by N-glycosidase activity. The treatment of HeLa cell cultures with abrin resulted in apoptotic cell death, as characterized by morphological and biochemical changes, i.e., cell shrinkage, internucleosomal DNA fragmentation, the occurrence of hypodiploid DNA, chromatin condensation, nuclear breakdown, DNA single strand breaks by TUNEL assay, and phosphatidylserine (PS) externalization. This apoptotic cell death was accompanied by caspase-9 and caspase-3 activation, as indicated by the cleavage of caspase substrates, which was preceded by mitochondrial cytochrome c release. The broad-spectrum caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (zVAD-fmk), prevented abrin-triggered caspase activation and partially abolished apoptotic cell death, but did not affect mitochondrial cytochrome c release. These results suggest that the release of mitochondrial cytochrome c, and the sequential caspase-9 and caspase-3 activations are important events in the signal transduction pathway of abrin-induced apoptotic cell death in the HeLa cell line.

Protection by Methanol Extract of Longan (Dimocarpus Longan Lour.) Peel against Kainic acid-Induced Seizure

  • Jo, Young-Jun;Eun, Jae-Soon;Kim, Hyoung-Chun;Cho, Hwang-Eui;Lee, Mi-Kyeong;Hwang, Bang-Yeon;Hong, Jin-Tae;Moon, Dong-Cheul;Oh, Ki-Wan
    • Natural Product Sciences
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    • v.16 no.2
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    • pp.99-106
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    • 2010
  • This experiment was undertaken to investigate whether methanol extract of fruit peel of Dimocarpus longan Lour. (MEFL) protects against kainic acid (KA)-induced seizure. Oral administration of MEFL (1, 2 and 4 mg/kg) increased KA (50 mg/kg)-induced survival rate and latency of convulsion onset, and deceased seizure scores and weight loss induced by intraperitoneal (i.p) injection of KA in mice. In addition, MEFL protected against cell death in the hippocampus of rat brain after KA-administration as analyzed by using TUNEL assay in rats. MEFL also significantly blocked seizure-form of electroencephalogram (EEG) power spectra induced by KA in rats. MEFL also inhibited elevation of [$Ca^{2+}$]i and increased [$Cl^-$]i induced by KA in cultured neuronal cells. Therefore, it is suggested that MEFL protects against seizure induced by KA, decreasing [$Ca^{2+}$]i.

What should be done for men with sperm DNA fragmentation?

  • Kim, Gi Young
    • Clinical and Experimental Reproductive Medicine
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    • v.45 no.3
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    • pp.101-109
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    • 2018
  • In an age when a small quantity of sperm can lead to pregnancy through in vitro fertilization or intracytoplasmic sperm injection, selecting healthy sperm is important. Sperm DNA fragmentation (SDF) is known to be higher in infertile men. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) and the alkaline comet test are SDF tests that directly measure DNA damage and have shown closer correlations with assisted reproduction results than indirect tools such as the sperm chromatin structure assay or the sperm chromatic dispersion test. It is difficult; however, to endorse a single test as the best test overall; instead, it is best to select a testing method based on each patient's clinical condition and goals. In a couple struggling with infertility, if the male partner has a high level of SDF, he should aim to decrease SDF through lifestyle modifications, antioxidant treatment, and ensuring an appropriate duration of abstinence, and physicians need to treat the underlying diseases of such patients. If sperm DNA damage continues despite the patient's and physician's efforts, other methods, such as micromanipulation-based sperm selection or testicular sperm extraction, should be used to select healthy sperm with nuclear DNA integrity.