• The Korea Journal of Herbology
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• v.27 no.3
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• pp.89-94
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• 2012

Objects : The purpose of this study was to observe the effect of Salviae Miltriorrhiza Radix(SMR) water-extract on intracerebral hemorrhage(ICH) and neuronal apoptosis in the injured areas. Method : ICH was induced by the stereotaxic intrastriatal injection of bacterial collagenase type IV in Sprague-Dawley rats. The rats were givened oral SMR treatment once a day for three days after the ICH treatment. TUNEL positive cells in the affected regions were performed by TUNEL assay, Bax and Bcl-2 positive cells by immunohistochemistry and the Bax expression by western blotting method. Results : The results are as follow; 1. SMR significantly reduced the number of TUNEL positive cells in the peri-hematoma reigions of ICH-induced rats. 2. SMR significantly reduced the number of Bax positive cells in the peri-hematoma regions of ICH-induced rats. 3. SMR did not affect the number of Bcl-2 positive cells in the peri-hematoma regions of ICH-induced rats. 4. SMR significantly reduced the Bax expressions compared with ICH group in hemorrhagic hemisphere of ICH-induced rats. Conclusion : These results suggest that SMR is effective in reducing neuronal apoptosis.

• Journal of Oriental Neuropsychiatry
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• v.21 no.2
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• pp.29-44
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• 2010

Objectives : The purpose of this study was to investigate the effect of Gentianae Radix on neurogenesis and apoptosis in ethanol- induced newborn rats hippocampus dentate gyrus. Methods : In vivo, laboratory animals were divided into three groups; Normal group(N), Control group(C) and Treated group (TG)(n=7 for each group). N were treated saline daily for five days. C were treated 1.5 g/kg ethanol and saline daily for five days. TG were treated 1.5 g/kg ethanol and 300 mg/kg Gentianae Radix daily for five days. BrdU(5-bromo-2-deoxyuridine) assay was used to test neurogenesis in the dentate gyrus. And TUNEL(Terminal deoxynucleotidyl transferase dUTP nick end labeling) assay was used to test apoptosis in the dentate gyrus. Three groups were measured body weight, serum ethanol concentration, BrdU-positive cells and TUNEL-positive cells in the dentate gyrus. In vitro, MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to test viability in SK-N-MC cells. BrdU assay was used to test neurogenesis in SK-N-MC cells. DNA fragmentation and caspase-3 enzyme activity assay were used to test apoptosis in SK-N-MC cells. And treated ethanol and Gentianae Radix of all in vitro tests were made various concentration. Results : In vivo, Gentianae Radix modulated ethanol-induced neurogenesis and apoptosis in newborn rats hippocampus dentate gyrus. In vitro, TG 100 ${\mu}g/ml$ have significantly modulated ethanol-induced neurogenesis and apoptosis in SK-N-MC cells. And only TG 100 ${\mu}g/ml$ have significantly protected SK-N-MC cells from ethanol-induced cytotoxicity. Conclusions : Gentianae Radix may have the effect that modulated ethanol-induced neurogenesis and apoptosis in SK-N-MC cells.

• The Journal of Korean Obstetrics and Gynecology
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• v.21 no.3
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• pp.18-33
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• 2008

Purpose: The research is to investigate the effect of TFE on apoptosis of human-derived breast cancer cells, to find out the relationship with apoptosis. Methods: Human-derived breast adenocarcinoma cell line, MCF-7 cells were treated by TFE with various concentration. The inducement effect of TFE on cell apoptosis was observed with MTT assay and the relationship between the treatment and apoptosis was investigated with FACS analysis, TUNEL assay and DNA laddering assay and the change in the protein levels of PARP and caspase-3 activities were also observed. The release of cytochrome-c was observed to find out the pathway of apoptosis induced by TFE. Results: The cell apoptosis was significantly induced in MCF-7 cells treated with TFE in concentration-dependent and time-dependent manner. It was verified by FACS analysis, TUNEL assay, DNA laddering assay that cell-death was caused not by necrosis but by apoptosis. The activity of PARP and caspase were increased concentration-dependently. The release of cytocrome-c was decreased in proportion to the concentration of the fruit extract. It therefore demonstrated that mitochondria were involved in apoptosis induced by TFE. The appearance of Bcl-2 protein was decreased concentration-dependently. Conclusion: The treatment by TFE induced apoptosis of human breast adenocarcinoma cell line, MCF-7. It seems likely that cell-death was caused by apoptosis and mitochondria were involved in it. The mechanism of protein change causing apoptosis seems related to the inhibition of Bcl-2 protein, the promotion of inversion from cytochrome-c into cytosol, the activation of caspase and the promotion of PARP cleavage.

• Korean Journal of Acupuncture
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• v.28 no.4
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• pp.17-28
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• 2011

Objectives : Burn is the important adverse effect of the moxibustion treatment. This research does with the purpose to utilize as basic data which the results of histological observation of the burn tissue which generates when operating various commercial indirect moxa (CIM) on the skin, evaluate the burn because of the surgical operation. Methods : It operated 6 kinds of CIM on the depilated abdomen of the rat. By using the H-E staining and TUNEL assay, the image was observed and analyzed. Results : It could be the moxibustion medical treatment using the CIM generated in the first degree burn until the third degree. And in some case, burn has not generated. By using the H-E stain, the second-degree burn and third degree was observed, and it was observed the first degree burn by using TUNEL assay. In the first degree burn, TUNEL reaction in the epidermal layer was confirmed. The damage to the dermal layer was observed in more than 2 degree burn. In the third degree, the tissue degeneration to the subcutaneous fat was observed but the thickness of the skin tissue was not observed. Conclusions : Basic data classifying the burn generated by the CIM treatment through the histological observation of the burn tissue caused by the CIM treatment were built according to this research result.

• Korean journal of applied entomology
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• v.55 no.3
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• pp.267-275
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• 2016

Chlorine dioxide has an insecticidal activity via its production of reactive oxygen species (ROS). Its cytotoxic activity has been regarded as a main cause of the insecticidal activity. This study tested a hypothesis that cytotoxicity of chlorine dioxide is resulted from its induction of apoptosis against target cells using ROS. Injection of chlorine dioxide significantly reduced total hemocyte counts of Plodia interpunctella larvae and subsequently killed the larvae. To analyze the cytotoxicity with respect to apoptosis, terminal deoxyribonucleotidyl transferase nick end translation (TUNEL) assay was performed. An insect cell line (Sf9) cells were exposed to different concentrations of chlorine dioxide. TUNEL assay showed that chlorine dioxide induced significant apoptosis of Sf9 cells in a dose-dependent manner. When different concentrations of chlorine dioxide were injected to larvae of P. interpunctella, it showed a dose-dependent induction of apoptosis against hemocytes. However, addition of vitamin E significantly suppressed the apoptosis induction and insecticidal activity of chlorine dioxide in a dose-dependent manner. These results suggest that cytotoxicity of chlorine dioxide is resulted from its induction of apoptosis against insect cells using ROS.

• Childhood Kidney Diseases
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• v.22 no.1
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• pp.22-27
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• 2018

Purpose: Podocytes are important architectures that maintain the crucial roles of glomerular filtration barrier functions. Despite this structural importance, however, the mechanisms of the changes in podocytes that can be an important pathogenesis of minimal change nephrotic syndrome (MCNS) are not clear yet. The aim of this study was to investigate whether apoptosis is induced by interleukin (IL)-13 in cultured human podocytes. Methods: Human podocytes were treated with different IL-13 doses and apoptotic cells were analyzed using terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL assay) and fluorescence-activated cell sorting (FACS). Results: The IL-13 increased the number of TUNEL-positive cells in a dose-dependent manner at 6 and 18 hours (P<0.05 and P<0.05, respectively). The apoptosis rate was appeared to be increased slightly in the IL-13-stimulated podocytes (8.63%, 13.02%, and 14.46%; 3, 10 and 30 ng/mL, respectively) than in the control cells (7.66%) at 12 hours by FACS assay. Conclusion: Our study revealed that IL-13 expression may increase podocyte apoptosis. Blocking the IL-13 signal pathway can potentially play an important role in regulating the apoptosis of podocytes.

• Journal of Acupuncture Research
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• v.29 no.1
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• pp.47-59
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• 2012

목적 : 이 연구는 봉독이 사람의 난소암 세포인 SKOV3와 PA-1에서 death receptor의 발현을 높여 세포자멸사를 촉진함으로써 암세포의 성장을 억제하는지 밝히고자 하였다. 방법 : 난소암의 세포자멸사의 관찰에는 DAPI, TUNEL staining assay를 시행하였으며, 세포자멸사 조절 단백질의 변동 관찰에는 western blot analysis를 시행하였고, 난소암 세포에서 death receptor의 변화를 관찰하기 위해 RT-PCR analysis를 시행하였다. 결과 : 1. DAPI, TUNEL staining assay 결과, 봉독은 투여량에 따라 세포자멸사의 유도를 통해 SKOV3와 PA-1 난소암세포의 증식을 억제하였고, 세포자멸사와 동반하여 DR4와 DR6의 발현이 두 암세포 모두에서 증가하였고, DR3의 출현은 PA-1 세포에서 증가하였다. 2. Death Receptor의 발현 증가에 따라 caspase-3, 8, 9 and Bax를 포함하는 세포자멸사 촉진 단백질의 발현이 동반하여 상승하였고 JAK2, STAT3의 인산화와 Bcl-2의 발현은 억제되었다. 3. siRNA 처리 시 봉독에 의한 DR3, DR4, DR6 발현증가와 STAT3의 활성억제가 역전되었다. 결론 : 이러한 결과는 봉독이 난소암 세포에서 DR3, DR4, DR6의 증가와 JAK2/STAT3 pathway의 억제를 통하여 세포자멸사를 유발한다는 것을 시사하며, 난소암의 예방과 치료에 효과적으로 활용될 수 있을 것으로 기대된다.

• Journal of Veterinary Clinics
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• v.23 no.2
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• pp.153-157
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• 2006

The purpose of this study was to determine the effects of mitomycin C on conjunctival fibroblast proliferation and apoptosis. Rabbit conjunctival fibroblast mono layers were treated with 48 hours application of mitomycin-C. The viability of cells was evauated by MTT assay. To examine the effect of mitomycin-C on the cell proliferation, immunocytochemistry for BrdU staning was performed. Then, we investigated whether mitomycin-C induced cell's apoptosis by TUNEL staining. As a result of MTT assay, the viability of cells was gradually inhibited in a dose dependent manner by mitomycin-C. The BrdU staining showed that mitomycin-C significantly suppressed the proliferation of conjunctival fibroblast at concentrations of 0.02% or more. When TUNEL assays were performed, the number of apoptotic cells increased 5.6-, 18.5-, and 33.8-fold compared with the control at 0.01, 0.02, and 0.04%, respectively, of mitomycin-C at 48 hours of exposure. Therefore, mitomycin-C may be useful as a regulator in the treatment of corneal diseases that manifest with scar formation and tumor of fibroblast expansion.

• Journal of Veterinary Clinics
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• v.24 no.3
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• pp.358-366
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• 2007

This study was performed to investigate the application of bovine amniotic membrane for canine corneal ulcer and to compare bovine amniotic membrane flap to nictitating membrane flap. Corneas were treated with 8.0mm trephine and 100% ethanol was applied and corneal epithelium was removed. This experiment was divided into three groups; control group, nictitating membrane flap group and amniotic membrane + nictitating membrane flap group. To compare corneal healing, fluorescein dye test, histopathological examination and TUNEL assay were performed. The healing size of the corneal epithelium in was largest among groups and it was significantly different with the other groups (P<0.05). Also, inflammatory cell deposits and keratocyte apoptosis in amniotic membrane flap group were smallest, but the width of the corneal epithelium was thickest among groups. Therefore, we tentatively predicted that the amniotic membrane flap would be an alternative effective method for the treatment of superficial canine corneal ulcer.

• Animal cells and systems
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• v.11 no.1
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• pp.23-31
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• 2007

Programmed cell death was characterized in the silkworm thoracic ganglia TG1, TG2 and TG3 during postembryonic periods by TUNEL assay. Apoptotic cells were detected in the three TGs of all larval stages except for day-1, 2 1st instar larvae, in which no apoptotic cells were found. From day-7 5th larva, the numbers of apoptotic cells were dramatically increased and peaked on day-1 pupa and day-2 pupa and then abruptly decreased. Apoptotic cells finally disappeared in day-1 adult. In-vivo injection of 20-hydroxyecdysone (20E) into day-8 5th larva resulted in a striking decrease of apoptotic cells. Actinomycin D (Act D) or cycloheximide (CHX), injected into hemolymph of day-8 5th larva, resulted in a decrease of apoptotic cells in the three TGs. Injection of caspase-8 and -3 inhibitors also blocked cellular apoptosis. These results will provide valuable information for understanding of cellular changes in the three TGs during metamorphosis of the insect species.