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Article in Chinese | WPRIM | ID: wpr-909552


Objective:To investigate the effects of fluoxetine (Flx) on lipidomics of hippocampal tissue in chronic unpredictable stress (CUS) model rats.Methods:A total of 30 Sprague-Dawley rats were randomly divided into Sham group, CUS group and CUS+ Flx group, with 10 rats in each group. Rats in the CUS group and CUS+ Flx group were received one or two random stimuli every day for 28 days, and then they were received intraperitoneal injection of normal saline(1 ml/kg) and fluoxetine(10 mg/kg) respectively once a day for 14 days. Rats in the Sham group were maintained in their home cages for 28 days, and then received intraperitoneal injection of saline (1 ml/kg) once a day for 14 days. The sugar water preference experiment was carried out 24 hours after the last injection, and then the rats were killed to separate the rat hippocampus. The levels of lipid composition in hippocampus were detected by high performance liquid chromatography-mass spectrometry. The relative content of lipid was analyzed by Simca-p 14.1 and LipidSearch software version 4.1. SPSS 19.0 was used for statistical analysis. One-way ANOVA or Kruskal-Wallis test was used for comparison among groups, and Bonferroni test was used for post-hoc test. Pearson correlation or Spearman correlation was used to analyze the correlation between behavioral indexes and lipid molecular level in hippocampus.Results:There was significant difference in sugar preference test among the three groups ( F=12.830, P<0.001). The percentage of sucrose intake of rats in CUS group ((43.57±12.38)%) was significantly lower than those in Sham group ((67.09±11.81)%) and CUS+ Flx group ((62.74±8.58)%) (both P<0.05). Ninety five differential lipid molecules were screened among the three groups by lipidomic analysis, mainly distributed in glycerophospholipids and sphingolipids. Among them, levels of PE (34∶1e)+ H( r=-0.477), PE(18∶1p/20∶1)+ H( r=-0.433), PE(18∶1/18∶1)+ Na( r=-0.603), PE(36∶2p)-H( r=-0.382), PE(16∶0/20∶4)-H( r=-0.464), PE(18∶0/18.2)-H( r=-0.482), PE(16∶0e/22∶6)-H( r=-0.514), PE(18∶1/20∶4)-H( r=-0.511) and CerG1 (d18∶2/24∶0+ O)+ H( r=-0.490) were negatively correlated with sucrose preference rate (all P<0.05), whereas levels of PE (42∶6p)+ Na( r=0.379), PE(34∶0p)-H( r=0.397) and SM (d22∶1/16.0)+ HCOO( r=0.388) were positively correlated with sucrose preference rate (all P<0.05). Conclusion:Flx improves the depressive-like behavior of CUS model rats, which may be related to the regulation of hippocampal glycerophospholipid and sphingolipid metabolism.

International Journal of Surgery ; (12): 95-98,封3, 2013.
Article in Chinese | WPRIM | ID: wpr-598165


Objective To explore the influence of HepG2 cells' proliferation and autophagy by the Nrf2-ARE pathway,and provide the experimental basis for clinical exploring effective liver cancer treatment.Methods Hepatic carcinoma HepG2 cells were cultured,and its proliferation inhibition rates and the change of cell cycle' s in each phase were explored by the MTT assay and flow cytometry.The hepatoma cells' autophagy was qualitative observed by inverted phase contrast microscope and fluorescence microscope.Results Inhibitory rate of HepG2 cells was obviously higher in the Nrf2 inhibitor BML-111 group than control group (P < 0.05),and the control group was aslo obviously higher than the Nrf2 inducer EGb group (P < 0.05).Flow cytometric analysis showed that G1 phase cells in the cell cycle increased,S phase cells reduced and G2/M period cells relatively increased in the Nrf2 inhibitor BML-111 group.But G1 phase cells reduced,S phase cells increased and G2/M period cells relative reduced in the Nrf2 inducer EGb group.Inverted phase contrast microscope and fluorescence microscope checked that ranging from the size of the bubble and autophagosome formed in Hepatoma HepG2 cytoplasmic of the Nrf2 inhibitor BML-111 group.Conclusions The Nrf2-ARE pathway played an reverse inhibition on HepG2 cells' proliferation and autophagy.After the inhibition of Nrf2-ARE pathway,HepG2 cells mostly stayed in the G1 phase of the cell cycle.

International Journal of Surgery ; (12): 849-852, 2012.
Article in Chinese | WPRIM | ID: wpr-429823


Hepatic carcinoma is one of the most common malignant tumors in China.Autophagy activity and the change of Nrf2-ARE pathway play an important role in the process of liver tumors.Nrf2 which is an important regulator to liver cancer belongs to the CNC family.Research discovered that after the inhibition of autophagy,Nrf2-ARE pathway activation contributes to the progression of hepatocellular carcinoma.This article summarizes the relationship between autophagy and the Nrf2-ARE pathway and its impact on the hepatic carcinoma.

Article in Chinese | WPRIM | ID: wpr-412578


Objective To investigate the effect of different amplification methods and probes with various length on the results of comparative genomic hybridization (CGH) analysis of pre-implanted single blastomere and to establish the basis for preimplantation genetic diagnosis.Methods Twenty blastomeres of embryo at 6-8 cells stage were randomly divided into A and B group with 10 in each.Twenty peripheral blood lymphocytes from a healthy man were similarly divided into C and D group with 10 in each.Degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR) was used to amplify whole genomic DNA in group A and C,and multiple displacement amplification (MDA) was used in group B and D for whole genome amplification (WGA).The specificity of resultant products was confirmed by amplification of TBX1 gene exon 2.CGH was performed respectively with 250-750 bp and 750-2000 bp probes prepared from the amplified whole genomic DNA.The result of CGH was verified by sex-determining region of Y (SRY).Results (1) Nine of the 10 samples in group A and all in group C were amplifiable by DOP-PCR,but there were multiple non-specific bands in the amplification of TBX1 exon 2 when WGA products were used as templates.When 250~750 bp probe was used in CGH,1 of the 5 blastomeres was failed and another one had different karyotype from that analyzed by SRY.(2) All samples in group B and D were successfully amplified by MDA,and the non-specific bands were significantly less in the amplification of TBX1 exon 2.All 5 blastomeres were successful in CGH with the 250~750 bp probe.Moreover,the karyotype was in agreement with that of SRY.(3) When 750 ~ 2000 bp probe was used,the CGH results were suboptimal.Conclusions In WGA of single blastomere,MDA is superior to DOP-PCR in the stability and specificity.The karyotype image detected by CGH with the 250~750 bp probe is clear and homogenous.