• Asian-Australasian Journal of Animal Sciences
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• v.16 no.3
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• pp.342-347
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• 2003

The enzyme $20{\alpha}$-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) catabolizes progesterone to $20{\alpha}$-dihydroprogesterone ($20{\alpha}$-OHP), and is appeared in rat corpora luteal and placenta. A polled samples of 10-15 placental and ovarian tissues collected from each individual rat were subjected to measurement of $20{\alpha}$-HSD activity. A $20{\alpha}$-HSD activity in the cytosol fraction was based on the generation of NADPH. In this study, it is designed to study cytosolic $20{\alpha}$-HSD activity in rat ovarian and placenta during pregnancy, and its relationship to embryonic mortality. It was found that from days 5 to 18 of pregnancy the $20{\alpha}$-HSD activities steady by decreased but at parturition time rapidly increased in ovary. On the other hand, placental cytosolic $20{\alpha}$-HSD activities were high detected from days 8 to 10 of pregnancy, not detectable from days 11 to 20 of pregnancy, but again very high at the time of parturition. Analysis of DEAE column chromatography revealed that two different types of $20{\alpha}$-HSD (HSD-1 and HSD-2) were found with similar activity in the placental cytosol on day 10 of pregnancy. The number of fetuses on day 10 of pregnancy was 15.4 and decreased significantly to 12.9 on day 12. The results suggested that expression of $20{\alpha}$-HSD in the placental tissues seems to be related the number of fetal survived in the specific time (days 11 and 12) which spontaneous fetal loss occurs.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.8
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• pp.3293-3299
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• 2015

Background: Arm protein lost in epithelial cancers, on chromosome X (ALEX) is a novel subgroup within the armadillo (ARM) family, which has one or two ARM repeat domains as opposed to more than six-thirteen repeats in the classical Armadillo family members. Materials and Methods: In the study, we explore the biological functions of ALEX1 in breast cancer cells. Overexpression of ALEX1 and silencing of ALEX1 were performed with SK-BR3 and MCF-7 cell lines. Cell proliferation and colony formation assays, along with flow cytometry, were carried out to evaluate the roles of ALEX1. Results: ALEX1 overexpression in SK-BR3 breast cancer cells inhibited proliferation and induced apoptosis. Furthermore, depletion of ALEX1 in MCF-7 breast cancer cells increased proliferation and inhibited apoptosis. Additional analyses demonstrated that the overexpression of ALEX1 activated the intrinsic apoptosis cascades through up-regulating the expression of Bax, cytosol cytochrome c, active caspase-9 and active caspase-3 and down-regulating the levels of Bcl-2 and mitochondria cytochrome c. Simultaneouly, silencing of ALEX1 inhibited intrinsic apoptosis cascades through down-regulating the expression of Bax, cytosol cytochrome c, active caspase-9, and active caspase-3 and up-regulating the level of Bcl-2 and mitochondria cytochrome c. Conclusions: Our data suggest that ALEX1 as a crucial tumor suppressor gene has been involved in cell proliferation and apoptosis in breast cancer, which may serve as a novel candidate therapeutic target.

• Microbiology and Biotechnology Letters
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• v.18 no.3
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• pp.239-243
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• 1990

In this study the authors investigated the distribution of cadmium accumulated in cadmium-iun tolerant Hansenula anomala B-7 cells and also the effect of cadmium on protein synthesis. 84.9% of the cadmium accumulated was distributed in the soluble fraction (cytosol, etc.). The intracellular protein content was decreased by cadmium (1,000 $\mu g$/ml), but the content of soluble protein preeipitated by ammonium sulfate (30-75% saturation) was increased compared with the content of it obtained from the cells grown without cadmium. Furthermore, in the cells grown with 1,000 $\mu g$/ml of cadmium t h higher molecular weight soluble protein was increased compared with the cells grown without caa, mium, but the lower molecular weight soluble protein was decreased. These results suggested that the protein synthesis was inhibited by cadmium, but synthesis of higher molecular weight soluble protein was remarkably stimulated by cadmium.

• Journal of Life Science
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• v.10 no.4
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• pp.347-353
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• 2000

This study was designed to investigate the effects of silkworm (Bombyx mori L.) powder (SWP) on oxygen radicals and their scavenger enzymes in liver membranes of rats. Sprague-Dawley(SD) male rats(160$\pm$10g) were fed basic diet (control group), and experimental diets(SWP-200 and SWP-400 groups) added 200 and 400 mg/kg BW/day for 6 weeks. Hydroxyl radical (.OH) levels resulted in a consistent decreases (4.0% and 7.2%, 5.0% and 14.1%, respectively) in liver mitochondria and microsomes of SWP-200 SWP-400 groups compared with control group, and O2 radical level was significantly decreased about 12% in liver cytosol of SWP-400 group compared with control group. Lipid peroxide(LPO) levels were significantly decreased (14.4% and 9.1%, respectively) in liver mitochondria and microsomes of SWP-400 group only compared with control group. Oxidized protein (OP) levels were remarkably decreased about 12.7% and 16.3% in liver microsomes only of SWP-200 and SWP-400 groups, but significant difference between liver mitochondria could not obtained. Mn-SOD activities were remarkably increased (15.8% and 25.2%, respectively) in mitochondria of SWP-200 and SWP-400 groups, but significant difference between Cu, Zn-SOD activities in these group could be not obtained. GSHPx activity was significantly increased in liver cytosol of SWP-400 group compared with control group. These results suggest that silkworm powder may play an effective role in a attenuating a oxidative stress and increasing a scavenger enzyme activity in liver membranes.

• Biomolecules & Therapeutics
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• v.15 no.3
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• pp.145-149
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• 2007

We have previously shown that 2,4,3',5'-tetramethoxystilbene (TMS), a synthetic trans-stilbene analogue acting as a potent inhibitor of human cytochrome P450 1B1, induces apoptotic cell death in human cancer cells. In the present studies, we report the mechanisms of apoptotic cell death by TMS in human promyelocytic leukemic HL-60 cells. We found that treatment of HL-60 cells with TMS suppressed the cell growth in a concentration-dependent manner with $IC_{50}$ value of about 0.8 ${\mu}M$. Immunoblot experiments revealed that DMHS-induced apoptosis was associated with cleavage of poly (ADP-ribose) polymerase. The release of cytochrome c from mitochondria into the cytosol was significantly increased in response to TMS. TMS caused activation of caspase-3 in a concentration-dependent manner and TMS-mediated caspase-3 activation was partially prevented by the caspase inhibitor, zVAD-fmk. Interestingly, we found that the cytotoxic effect of anticancer drugs such as paclitaxel, docetaxel, or etoposide was enhanced in the presence of TMS. Simultaneous treatment with TCDD also significantly increased cytotoxic effects of TMS alone or TMS and anti-cancer agents. Taken together, our present results indicated that TMS leads to apoptotic cell death in HL-60 cells through activation of caspase-3 activity and release of cytochrome c into cytosol. The ability of TMS to increase cytotoxic effect of anticancer drugs may contribute to its usefulness for cancer chemotherapy.

• Korean Journal of Medicinal Crop Science
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• v.11 no.2
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• pp.148-154
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• 2003

This study was designed to investigate the effect of Korean Red ginseng(Panax ginseng C.A. Meyer) extracts on antioxidative activities in Sprague-Dawley rats. This study also evaluated the synergistic effect of Korean red ginseng(RG) extracts with Saururus chinensis(Lour.) Baill(SC). and Rubus coreanus Miq.(RC) extracts. Experimental groups were divided into supplementation type(RG extracts, RG with SC and RC extracts) and amounts of extracts. Rats were received drinking water with or without RG, RC and SC extracts for eight weeks. In the antioxidant enzyme activities of liver cytosol, superoxide dismutase, catalase and glutathione peroxidase activities were significantly increased in RG groups and RG with SC and RC groups compared to control group. The antioxidative activities were increased in proportion to supplementation period and amounts of extracts. These results suggest that RG, RC and SC extracts have an beneficial effect to enhance the cellular antioxidant activities in rats.

• Development and Reproduction
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• v.19 no.4
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• pp.227-234
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• 2015

The objective of this study was to investigate the expression of bovine luteum expressed sequence tags (ESTs), vascular endothelial growth factor (VEGF), and tumor necrosis factor receptor 1 (TNFR1) and the presence of functional ESTs in the bovine corpus luteum (CL) during different stages of the estrus cycle. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed a difference in the expression of ESTs during the CL stage. Concentration of ESTs in the CL tissue increased significantly from the mid-luteal stage and decreased thereafter. RT-PCR analysis showed higher levels of the EST genes in the CL of the mid-luteal stage than in other stages, and the same level of expression of VEGF. Immunohistochemistry analysis of the tissue from CL formation to regression showed low cytosol and aggregation of the nucleus. And activity caspase 3 (apoptosis detector) was most strongly detected in the CL1 stage of bovine. During the estrous cycle, the cytosol was magnified and differentiation of the nucleus was clearly manifested. The ESTs affected the CL, and the relationship between VEGF and TNFR1 played a pivotal role for CL development and activation, dependent on the stage of CL. These results suggest local production of ESTs, the presence of functional ESTs in the bovine CL, and that ESTs play a role in regulating the function of cell death in bovine CL.

• Archives of Pharmacal Research
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• v.26 no.10
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• pp.826-831
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• 2003

ADP-ribose pyrophosphatase (ADPRase) hydrolyzes ADP-ribose (ADPR) into AMP and ribose-5'-phosphate. It is classified into two groups, $Mg^{2+}$-dependent and $Mg^{2+}$-independent ADPRase, depending on its $Mg^{2+}$requirement. Here, we purified $Mg^{2+}$-dependent ADPRase from rabbit liver and examined what factors affect $Mg^{2+}$ requirement. The purified enzyme showed a single band with the molecular weight of 34 kDa on SDS-PAGE both in the presence and absence of 2-mercaptoethanol. The molecular weight of the native enzyme calculated by gel filtration was 68 kDa, indicating that ADPRase is a dimer made up of two identical subunits. $Mg^{2+}$-dependent ADPRase with the highest ADPR affinity had a $K_m$ of 160$\pm$10 $\mu$M and a pH optimum of around pH 9.5. Treatment of the purified ADPRase with heated cytosol fractions at 37$^{\circ}C$ for 3 h caused some changes in the chemical properties of the enzyme, including an increase in molecular weight, a decrease in solubility, and a loss of $Mg^{2+}$-dependency. The molecular weight of the cytosol-treated ADPRase measured by gel filtration was over 420 kDa, suggesting, for the first time, that ADPRase could be polymerized by undefined cytoplasmic factors, and that polymerization is accompanied by changes in the solubility and metal ion dependency of the enzyme.

• The Journal of Internal Korean Medicine
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• v.22 no.3
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• pp.309-320
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• 2001

Objectives : This study was done to investigate the protective effects of Samgiinjin-tang on liver injury of rats induced by CCI4 and d-galactosamine. Methods: All animals were divided into .5 groups, those were normal group(untreated), control group(treated with 0.9% Saline solution), sample I group(2,250mg/kg administrated), sample II group(4,500mg/kg administrated), Silymarin 200mg/kg administrated group. Liver injury of rats were induced by CCI4 and d-galactosamine, and then the serum transaminases(ALT&AST) alkaline phosphatase(ALP), lactic dehydrogenase(LDH) for enzyme activities, liver weight, lipid peroxidation and catalase, glutathione S-transferase(GST) for enzyme activities were measured. Results : The inhibitory effects on the serum ALT, AST activities in liver injury of rats induced by CCI4 were noted in both sample I and sample II group. The inhibitory effects on the serum ALP, LDH activities and the Lipid peroxidation of Mitochondria & Cytosol were noted in only sample II group. The decreased effects on the GST activities of Homogenate & Cytosol were inhibited in both sample I and sample II groups. The decreased effects on the GST activities of Mitochondria & Microsome were inhibited in sample II group. The inhibitory effects of the serum ALT, AST, LDH activities in liver injury of rats induced by d-galactosamine were noted in both sample I and sample II groups. In serum AST activities, sample II group. Conclusions : Samgiinjin-tang has protective effects against liver injury of rats induced by CCI4 and d-galactosamine. So it is required to study about the actions of mutual relation of medicines and patho-mechanism by experiment.

• The Journal of Internal Korean Medicine
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• v.30 no.1
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• pp.51-63
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• 2009

Background and Objective : The prospects for developing an anti-apoptotic natural component or a compound that exerts a neuroprotective effect with few or no side effects for the treatment of neurodegenerative disease appear favorable. In the present study, we evaluated the effects of the methanol extract of Phellodendri Cortex (PC extract) on 1-methyl-4-phenyl pyridinium($MPP^+$)-induced apoptosis in PC-12 cells. Materials and Methods : We used the methanol extract of Phellodendri Cortex (PC extract). PC-12 cells were cultured by RPMZ-1640. We found the PC extract's gene expression (Bax, Bcl-2) by using RT-PCR. We examined the PC extract's protein expression (Bcl-2, Bax, cytochrome c, poly (ADP-ribose) polymerase (PARP), caspase-3) by SDS-PAGE and Western blot. Results : Apoptosis in $MPP^+$-induced PC-12 cells was accompanied by an increased Bax/Bcl-2 ratio, release of cytochrome c to the cytosol and the activation of caspase-3. PC extract inhibited the down-regulation of Bcl-2 and the up-regulation of Bax, as well as the release of mitochondrial cytochrome c into the cytosol. In addition, PC extract attenuated caspase-3 activation and cleavage of poly (ADP-ribose) polymerase (PARP). Conclusion : These results suggest that the neuroprotective potentials of PC extract against $MPP^+$-induced apoptosis can be. at least partially, ascribed to its anti-apoptotic effects in PC-12 cells.