• Title/Summary/Keyword: BAX

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Effects of EA Application to GV20 and LI4 on BAX and NF-kB Expression of the SD-Rat's Hippocampus with Global Ischemia

  • Choi, Jung-Hyun;Kim, Sung-Won;Lee, Jae-Gap;Kim, Min-Hee;Kim, Ji-Sung;Choi, Yoo-Rim;Yun, Young-Dae;Kim, Chi-Hyok;Kim, Yong-Seong;Kim, Nyeon-Jun;Lee, Ju-Hwan;Lee, Sang-Bin
    • Journal of International Academy of Physical Therapy Research
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    • v.1 no.2
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    • pp.136-142
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    • 2010
  • This study aims to reveal how EA affects BAX and NF-kB involved in cell deaths from global ischemia, and to do this, observes the changes of BAX and NF-kB caused by EA application after transient global ischemia. The experimental method is to give rise to global ischemia and apply EA to 27 SD rats with the particulars of being six-week-old, male, around-300 gram-weighing, and adapted to laboratory environment for more than a week, and divide them into three groups, that is, GV20 EA group(n=9), L14 EA group(n=9), no-treatment GI group(n=9), and then observe their changes of BAX and NF-kB at the time lapse of 6 hours, 9 hours and 12 hours after ischemia, using western blotting. The numerical decrease of BAX expression at the time lapse of 9 hours after EA application, though not statistically significant, was observed in GV20 EA group and L14 EA group, and the NF-kB expression appeared statistically significant decrease in GV20 EA group and L14 EA group, but the expression was higher in the group with EA application. Therefore, EA application at the early phase of global ischemia is considered to affect BAX and NF-kB and play a positive role in decreasing apoptosis and cell deaths by inflammation.

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Effects of Sunghyangjungki-san(Xingxiangzhengqi-san) on Bax and Bcl-2 Expressions in the MCAO Rats (성향정기산(星香正氣散)이 중대뇌동맥폐쇄 흰쥐의 Bax 및 Bcl-2 발현에 미치는 영향)

  • Jeong, Beoul;Lee, Jong-Soo
    • Journal of Korean Medicine Rehabilitation
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    • v.18 no.2
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    • pp.33-43
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    • 2008
  • Objectives : Sunghyangjungki-san(Xingxiangzhengqi-san) is a herb decoction prescribed frequently for stroke patients. The present study investigated neuroprotective effects of Sunghyangjungki-san(Xingxiangzhengqi-san) against the ischemic damage of the rat brain. Neuronal cell death under the cerebral ischemia is distinguished with the delayed cell death through apoptosis. Consequently, the effects of Sunghyangjungki-san(Xingxiangzhengqi-san) was evaluated with Bax and Bcl-2 expressions as apoptosis related factors in the brain tissues. Methods : The ischemic damage was induced by the middle cerebral artery occlusion(MCAO) method in Sprague-Dawley rats. Water extract of Sunghyangjungki-san(Xingxiangzhengqi-san) was treated for 5 days after the MCAO. Neurological scores and infarct size with TTC were measured. Bax and Bcl-2 expressions in the brain tissues were observed with immunohistochemistry. Results : Sunghyangjungki-san(Xingxiangzhengqi-san) treatment improved neurological score significantly at 5 days after the MCAO. Sunghyangjungki-san(Xingxiangzhengqi-san) treatment decreased infarct size by the MCAO, but it was not significant statistically. Sunghyangjungki-san(Xingxiangzhengqi-san) treatment attenuated Bax positive neurons significantly in the cerebral penumbra and the caudate putamen. Bcl-2 positive neurons were increased, but not significant. Sunghyangjungki-san(Xingxiangzhengqi-san) treatment increased Bcl-2/Bax expression ratios significantly in the cerebral penumbra and the caudate putamen. Conclusions : These results suggest that Sunghyangjungki-san(Xingxiangzhengqi-san) has a neuroprotective effect on the stroke with modulation of apoptosis related factors.

Functional Screening for Cell Death Suppressors and Development of Multiple Stress-Tolerant Plants

  • Moon Hae-Jeong;Baek Dong-Won;Lee Ji-Young;Nam Jae-Sung;Yun Dae-Jin
    • Journal of Plant Biotechnology
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    • v.5 no.3
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    • pp.143-148
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    • 2003
  • Bax, a mammalian pro-apoptotic member of the Bcl-2 family induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorhodamine123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MSP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to playa novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.

Functional Screening for Cell Death Suppressors and Development of Multiple Stress-Tolerant Plants

  • Moon, Hae-Jeong;Baek, Dong-Won;Lee, Ji-Young;Nam, Jae-Sung;Yun, Dae-Jin
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2003.04a
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    • pp.65-71
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    • 2003
  • Bax, a mammalian pro-apoptotic member of the Bcl-2 family, induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorho-damine 123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MBP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to play a novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.

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Tumor Suppressor Protein p53 Promotes 2-Methoxyestradiol-Induced Activation of Bak and Bax, Leading to Mitochondria-Dependent Apoptosis in Human Colon Cancer HCT116 Cells

  • Lee, Ji Young;Jee, Su Bean;Park, Won Young;Choi, Yu Jin;Kim, Bokyung;Kim, Yoon Hee;Jun, Do Youn;Kim, Young Ho
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1654-1663
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    • 2014
  • To examine the effect of tumor suppressor protein p53 on the antitumor activity of 2-methoxyestradiol (2-MeO-$E_2$), 2-MeO-$E_2$-induced cell cycle changes and apoptotic events were compared between the human colon carcinoma cell lines HCT116 ($p53^{+/+}$) and HCT116 ($p53^{-/-}$). When both cell types were exposed to 2-MeO-$E_2$, a reduction in the cell viability and an enhancement in the proportions of $G_2/M$ cells and apoptotic sub-$G_1$ cells commonly occurred dose-dependently. These 2-MeO-$E_2$-induced cellular changes, except for $G_2/M$ arrest, appeared to be more apparent in the presence of p53. Immunofluorescence microscopic analysis using anti-${\alpha}$-tubulin and anti-lamin B2 antibodies revealed that after 2-MeO-$E_2$ treatment, impaired mitotic spindle network and prometaphase arrest occurred similarly in both cell types. Following 2-MeO-$E_2$ treatment, only HCT116 ($p53^{+/+}$) cells exhibited an enhancement in the levels of p53, p-p53 (Ser-15), $p21^{WAF1/CIP1}$, and Bax; however, the Bak level remained relatively constant in both cell types, and the Bcl-2 level decreased only in HCT116 ($p53^{+/+}$) cells. Additionally, mitochondrial apoptotic events, including the activation of Bak and Bax, loss of ${\Delta}{\psi}m$, activation of caspase-9 and -3, and cleavage of lamin A/C, were more dominantly induced in the presence of p53. The Bak-specific and Bax-specific siRNA approaches confirmed the necessity of both Bak and Bax activations for the 2-MeO-$E_2$-induced apoptosis in HCT116 cells. These results show that among 2-MeO-$E_2$-induced apoptotic events, including prometaphase arrest, up-regulation of Bax level, down-regulation of Bcl-2 level, activation of both Bak and Bax, and mitochondria-dependent caspase activation, the modulation of Bax and Bcl-2 levels is the target of the pro-apoptotic action of p53.

Effects of 8-week Exercise on Bcl-2, Bax, Caspase-8, Caspase-3 and HSP70 in Mouse Gastrocnemius Muscle (8주간 운동이 생쥐의 gastrocnemius에서 Bcl-2, Bax, caspase-8, caspase-3와 HSP70에 미치는 영향)

  • Kim, Ki-Bum;Kim, Yong-An;Park, Jung-Jun
    • Journal of Life Science
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    • v.20 no.9
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    • pp.1409-1414
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    • 2010
  • The aim of this study was to investigate the effects of exercise on intrinsic and extrinsic apoptosis signaling pathways in skeletal muscle. ICR-type white male mice were divided into a control group (CON: n=10) and an exercise training group (EX: n=10) after a 1 week adaptation period. EX performed treadmill running at 16.4 m/min with a 4% incline, 40 min/day and 5 days/week for 8 weeks. Cervical dislocation was performed at 48 hours after the last bout of exercise, after which gastrocnemius skeletal muscles were immediately collected. The results of verifying the intrinsic apoptosis pathway showed that there were no significant differences in Bcl-2, Bax, or the ratio of Bax/Bcl-2 proteins between EX and CON. On the other hand, the results of verifying the extrinsic apoptosis pathway showed that caspase-8 proteins were significantly lower in EX than in CON (p<0.05). Apoptosis suppressing protein HSP70 was higher in EX than in CON. In addition, caspase-3, which is the final factor for apoptosis, was not activated. These results indicate that apoptosis did not develop since caspase-3 is non-cleaved by the effects of caspase-8 and HSP70 extrinsic pathways rather than Bcl-2 and Bax intrinsic pathways among signal pathways for apoptosis.

Effects of Salvia Miltriorrhiza Radix on Neuronal Apoptosis following Intracerebral Hemorrhage of Rats (단삼(丹蔘)이 뇌조직출혈 흰쥐의 신경세포 자연사에 미치는 영향)

  • Lee, Hung-Gi;Kim, Youn-Sub
    • 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.

Effect of Gastrodiae Rhizoma on Apoptosis in Cerebral Infarction Induced by Middle Cerebral Artery Occlusion in Rats (천마가 중대뇌동맥 폐쇄 흰쥐의 신경세포 자연사에 미치는 영향)

  • Youn, You-Suk;Lee, Jong-Soo
    • Journal of Korean Medicine Rehabilitation
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    • v.19 no.3
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    • pp.1-13
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    • 2009
  • Objectives : This study evaluates neuroprotective effect of Gastrodiae Rhizoma on apoptosis in the cerebral infarct. Methods : Cerebral infarct was induced by the middle cerebral artery occlusion (MCAO) for 2 hours with intraluminal thread method in Sprague-Dawley rats. Then ethanol extract of Gastrodiae Rhizoma was administered orally for 3 days. Infarct area and volume were evaluated with TTC staining. Neuronal apoptosis was identified with TUNEL labeling. Apoptosis modulatory effect was observed with immunohistochemical Bax, Bcl-2, iNOS, and MMP-9 expressions in penumbra. Results : 1. Ethanol extract of Gastrodiae Rhizoma reduced infarct size partly and volume significantly of in the MCAO rat brain. 2. Ethanol extract of Gastrodiae Rhizoma reduced TUNEL positive cell ratio in the penumbra of MCAO rat brain significantly. 3. Ethanol extract of Gastrodiae Rhizoma suppressed Bax, iNOS and MMP-9 expression in the penumbra of MCAO rat brain significantly. 4. Ethanol extract of Gastrodiae Rhizoma did not change Bcl-2 expression in the penumbra of MCAO rat brain. But expression ratio of Bcl-2 against Bax was increased in the Gastrodiae Rhizoma group. Conclusions : These results suggest that Gastrodiae Rhizoma plays an anti-apoptotic neuroprotective effect through suppression of Bax, iNOS, and MMP-9 expressions and relative up-regulation of Bcl-2 in the ischemic brain tissue.

Mangiferin Induces Apoptosis by Regulating Bcl-2 and Bax Expression in the CNE2 Nasopharyngeal Carcinoma Cell Line

  • Pan, Li-Li;Wang, Ai-Yan;Huang, Yong-Qi;Luo, Yu;Ling, Min
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7065-7068
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    • 2014
  • To investigate the anti-proliferative mechanism of mangiferin in a human nasopharyngeal carcinoma cell line, CNE2 cells were incubated with different concentrations of mangiferin (12.5, 25, 50, 100, 150 and $200{\mu}M$) or with PBS as a control for 72 hours. Analyses were made of the cell cycle and apoptosis with measurement of mRNA and protein levels of two apoptosis-related genes, Bcl-2 and Bax. Flow cytometry assays showed mangiferin could inhibit CNE2 cell proliferation via G2/M arrest and induction of early apoptosis. Real time PCR and Western blotting showed the mRNA and protein level of Bcl-2 to be down-regulated, while those of Bax were upregulated, when CNE2 cells were treated with mangiferin. This investigation indicated anti-proliferation effects of mangiferin through induction of cell apoptosis regulated by Bcl-2 and Bax expression.

Curcumin Inhibits Human Non-small Cell Lung Cancer A549 Cell Proliferation Through Regulation of Bcl-2/Bax and Cytochrome C

  • Li, Yue;Zhang, Shuai;Geng, Jian-Xiong;Hu, Xiao-Yang
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.8
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    • pp.4599-4602
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    • 2013
  • We intended to study the mechanism of the inhibitory action of curcumin on human non-small cell lung cancer A549 cell. The cell growth was determined by CCK-8 assay, and the results indicated that curcumin inhibited the cell proliferation in a concentration dependent manner. And to further confirm the relative anti-cancer mechanism of curcumin, RT-PCR was carried out to analysis the expression of relative apoptotic proteins Bax, Bcl-2. We found that curcumin could up-regulate the expression of Bax but down-regulate the expression of Bcl-2 in A549 cells. In addition, curcumin affect the mitochondrial apoptosis pathway. These results suggested that curcumin inhibited cancer cell growth through the regulation of Bcl-2/Bax and affect the mitochondrial apoptosis pathway.