ABSTRACT
ETS-1 is a transcription factor that is a member of the E26 transformation-specific (ETS) family. Galanin receptor 2 (GalR2), a subtype of receptors of the neuropeptide galanin, has been shown to have an antidepressant-like effect after activation in rodents. Our previous study has shown that overexpression of ETS-1 increases the expression of GalR2 in PC12 phaeochromocytoma cells. However, whether ETS-1 has an antidepressant-like effect is still unclear. In this study, we found that chronic mild stress (CMS) decreased the expression of both ETS-1 and GalR2 in the ventral hippocampus of rats. Meanwhile, we demonstrated that overexpression of ETS-1 increased the expression of GalR2 in primary hippocampal neurons. Importantly, we showed that overexpression of ETS-1 in the ventral hippocampus counteracted the depression-like behaviors of CMS rats. Furthermore, we found that overexpression of ETS-1 increased the level of downstream phosphorylated extracellular signal-regulated protein kinases 1 and 2 (p-ERK1/2) of GalR2 in the ventral hippocampus of CMS rats. Taken together, our findings suggest that ETS-1 has an antidepressant-like effect in rats, which might be mediated by increasing the level of GalR2 and its downstream p-ERK1/2 in the ventral hippocampus.
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@#Objective To construct dual-luciferase reporter plasmids containing the wild type and mutant rat extracellular signal-regulat-ed kinase 1 (ERK1) gene 3' untranslated regions (UTR) which were used to detect rno-miR-15b-5p's putative target gene. Methods The rat ERK1 gene 3' UTR fragment was amplified by polymerase chain reaction (PCR) from PC12 cell cDNA and cloned into pmiR-RB-ReportTM vector. The mutant rat ERK1 gene 3' UTR fragment was obtained by overlap PCR and inserted into pmiR-RB-ReportTM vector. Successful wild type and mutant recombinant plasmids were confirmed by DNA sequencing. PC12 cells were co-transfected with rno-miR-15b-5p mim-ic and pmiR-ERK13' UTR or pmiR-ERK1-mut 3' UTR and then analyzed by dual-luciferase reporter assay system. The achieved mutation sequence of the target site TGCTGCT was mutated to CGAACGT and GTACACG, respectively. Results The wild-type reporter vector pmiR-ERK13' UTR and the mutant reporter vector pmiR-ERK1-mut 3' UTR were successfully constructed. The rno-miR-15b-5p mimic de-creased the activity of pmiR-ERK13' UTR plasmid (P<0.001) but did not decrease the activity of pmiR-ERK1-mut 3' UTR plasmid. Conclu-sion The recombinant pmiR-ERK13' UTR and pmiR-ERK1-mut 3' UTR plasmids were constructed successfully, and luciferase activities demonstrated that the 3' UTR of ERK1 gene might be a potential target of rno-miR-15b-5p.
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Objective To explore the curative effect of deep-brain magnetic stimulation (DMS) on learned helplessness behavior in the chronic restrained stress (CRS) rat model. Methods Twenty-nine Sprague-Dawley rats were randomly divided into control group (n=8) and CRS group (n=21). CRS group was exerted chronic restrained stress, while the control group did not receive any stress, for three weeks. Then learned helplessness behavior was tested using Forced Swimming Test (FST) and the hopeless rats of the CRS group were divided ran-domly into sham group (n=6), DMS group (n=8) and citalopram group (n=7), that received corresponding treatment respectively. They were evaluated with FST again after one-week treatment. Results The immobile time in FST was longer in CRS group than in the control group after three-week stress (F=11.260, P=0.002). After one-week treatment, no significant improvement was found in the citalopram group (F=1.565, P=0.235), however, the immobile time in DMS group decreased (F=6.277, P=0.025), and was shorter than that in the sham group (F=5.560, P=0.036). Conclusion CRS could result in learned helplessness behavior, which could be alleviated with one-week DMS.
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Objective To explore the effect of intracerebroventricular injection of AR-M1896, a galanin receptor 2 agonist, on depres-sion-like behavior in rat chronic mild stress (CMS) model. Methods 48 Sprague-Dawley rats were randomly assigned into control group, CMS group, CMS artificial cerebrospinal fluid (aCSF) group and CMS AR-M1896 group equally. The control group received no interven-tion, and the other groups were established chronic mild stress model. After six-week of stress, forced swim test and sucrose preference test were conducted to identify the CMS rats. AR-M1896 or aCSF was injected into the lateral ventricle of CMS AR-M1896 group and CMS aC-SF group, respectively. The immobility time and climbing time in the forced swim test were analysed, and the sucrose consumption percent-age in the sucrose preference test was measured. Results The immobility time decreased (F=11.998, P<0.01), climbing time increased (F=8.268, P<0.05), and the sucrose consumption percentage increased (F=10.352, P<0.01) in CMS AR-M1896 group, compared with CMS aC-SF group. Conclusion Intracerebroventricular administration of galanin receptor 2 agonist AR-M1896 is effective on depression in CMS model rats.
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One pair of primers were designed and synthesized based on the cDNA sequence encoding Homo sapiens poly (ADP-ribose) polymerase family, member 10 (PARP10) reported on the GenBank. The cDNA sequence encoding PARP10 was cloned from 293FT cell by RT-PCR. Then the RT-PCR product was cloned into pCMV-Myc and pEGFP-C1 plasmids. The interaction between PARP10 and beta-actin was identified through immuno-precipitation and laser confocal microscopy. Extensive expression of PARP10 in mouse tissues was confirmed by RT-PCR. Besides, Western blotting analysis indicated that cell injury caused by UV treatment could promote the expression of PARP10. The results in this paper would benefit further study of PARP10.
Subject(s)
Animals , Humans , Mice , Actins , Metabolism , Poly(ADP-ribose) Polymerases , Metabolism , Radiation Effects , Protein Interaction Domains and Motifs , Proto-Oncogene Proteins , Metabolism , Radiation Effects , Stress, Physiological , Radiation Effects , Tissue Distribution , Ultraviolet RaysABSTRACT
NS1 is a non-structural protein of the influenza A virus, which could only be expressed when cells are infected. The effect of NS1 protein on host cell is still not clear. To understand the role of NS1 protein in cell infection, recombinant plasmid pCMV-myc-NS1 was constructed, and then transfected into A549 cells. Two-dimensional electrophoresis was employed to analyze proteins regulated by NS1 that could reflect the interaction between influenza virus and host cells at the protein level. The influence of NS1 on cell proliferation and cell cycle was also studied. The result showed that not only could NS1 remarkably affect metabolism, but it could also slow down cell proliferation through blocking cell cycle.
Subject(s)
Animals , Humans , Mice , 3T3 Cells , Cell Proliferation , Influenza A virus , Genetics , Protein Biosynthesis , Transfection , Viral Nonstructural Proteins , Genetics , PhysiologyABSTRACT
AIM: To compare the contents of calycosin and fermononetin between Radix Astragali and its fried product with honey. METHODS: The chromatographic column was a Waters SunfireTM C18 column(250 mm ? 4. 6 mm,5 ?m). The mobile phase was the mixture of methanol and water with a linear gradient elution at a flow rate of 1 mL/min. The ultraviolet wavelength was set up 254 nm,and the column temperature was maintained at 35 ℃. RESULTS: The contents of calycosin and fermononetin in Radix Astragali were 0. 247 9 mg/g and 0. 125 2 mg/g respectively,the contents of calycosin and fermononetin in Radix Astragali fried with honey were 0. 225 6 mg/g and 0. 108 2 mg/g,respectively. CONCLUSION: The contents of calycosin and fermononetin in Radix Astragali fried with honey decreased.