Effects of prenatal maternal cold stress on spontaneous, exploratory and anxious behaviors in offspring / 中国应用生理学杂志

OBJECTIVE@#To study the effects of prenatal cold stress on the behavior and mood of offspring in pregnant rats.@*METHODS@#Six SPF-class Wister pregnant rats were randomly divided into normal temperature control group and cold stress group with 3 rats in each group. The pregnant female rats in the normal temperature control group were kept in the environment of (22 ±2)℃, and the pregnant female rats in the cold stress group were placed in the artificial intelligence climate chamber at(4 ±0.1)℃ for 7 days before the birth, and the young rats were divided into normal temperature after the young rats were born. After the young rats were born, they were divided into normal temperature control group of male rats (MR, 22), normal temperature control group of mother rats (FR, 15), cold stress group of male rats (MC, 15), and cold stress group of female rats (FC, 15) .In the fourth generation of the offspring, the open field experiment and the elevated cross maze test were carried out.@*RESULTS@#In the open field experiment, there was no significant difference in spontaneous activity and exploration behavior between the normal temperature control group and the cold stress group (P＞0.05). In the elevated plus maze experiment, the retention time of the open arms, the number of open arms and the distance of the male and female rats in the cold stress group were significantly higher than those in the normal temperature control group (P＜0.05).@*CONCLUSION@#Prenatal maternal cold stress has no significant effect on spontaneous activity, exploration behavior and activity level of offspring, but the offspring have obvious abnormal behaviors with reduced anxiety behavior.

Biological function prediction of mir-210 in the liver of acute cold stress rat / 生理学报

The study was aimed to observe mir-210 expression in liver tissue of acute cold stress rat and predict the function of mir-210 in cold stress. Thirty SPF Wistar male rats which were 12-week-old and weighed (340 ± 20) g were used. The rats were pre-fed in normal room temperature for one week, and then were randomly divided into acute cold stress group at (4 ± 0.1) °C and normal control group at (24 ± 0.1) °C. After the rats were treated with cold stress for 12 h, the liver tissue was extracted and the gene expression of mir-210 was assayed using qRT-PCR. The results demonstrated that the gene expression of mir-210 was significantly enhanced in acute cold stress group compared with that in normal control group (n = 3, P < 0.01). The bioinformatics analysis showed that mir-210 has over hundreds of target genes and four kinds of target genes such as E2F3, RAD52, ISCU and Ephrin-A3 are more relative with liver cold stress. ISCU regulates the cell respiratory metabolism and Ephrin-A3 is related with cell proliferation and apoptosis. On the other hand, up-regulated mir-210 affects the DNA repairing mechanism which usually leads to genetic instabilities. Our results suggest that cold stress-induced up-regulation of mir-210 in liver harmfully influences cell growth, energy metabolism and hereditary.

Cold inducible RNA-binding protein inhibits hippocampal neuronal apoptosis under hypothermia by regulating redox system / 生理学报

In this study, we intend to confirm our hypothesis that cold inducible RNA-binding protein (CIRP) can inhibit neuronal apoptosis through suppressing the formation of oxygen free radicals under hypothermia. Primary rat hippocampal neurons were isolated and cultured in vitro, and were divided into five groups: (1) normal control group (37 °C), (2) cells infected by empty viral vector group, (3) CIRP over-expressed group, (4) CIRP knock-down group, and (5) hypothermia control group. Cells in groups 2-5 were cultured under 32 °C, 5% CO2. Apoptosis of hippocampal neurons were detected by Annexin V-FITC/PI staining and flow cytometry; Expression of CIRP was determined by Western blot; Redox-related parameters (T-AOC, GSH-Px, SOD, MDA) were detected by ELISA kits. Results showed that CIRP expression levels were significantly increased (P < 0.01) and the apoptotic rates were significantly decreased (P < 0.01) in hypothermia control group and CIRP over-expressed group when compared with normal control group. On the other hand, the apoptotic rate was significantly increased (P < 0.05) in CIRP knock-down group compared with that in hypothermia control group. The levels of redox parameters in hypothermia control group and CIRP over-expressed group were significantly changed in comparison with those in normal control group, CIRP knock-down group and empty viral vector infected group, respectively (P < 0.05 or P < 0.01). These results suggest that up-regulation of CIRP by hypothermia treatment can protect the neuron from apoptosis through suppressing the formation of oxygen free radicals.

Screening differentially expressed plasma proteins in cold stress rats based on iTRAQ combined with mass spectrometry technology / 中国应用生理学杂志

<p><b>OBJECTIVE</b>Isobaric tags for relative and absolute quantitation (iTRAQ) combined with mass spectrometry were used to screen differentially expressed plasma proteins in cold stress rats.</p><p><b>METHODS</b>Thirty health SPF Wistar rats were randomly divided into cold stress group A and control group B, then A and B were randomly divided into 3 groups (n = 5): A1, A2, A3 and B1, B2, B3. The temperature of room raising was (24.0 +/- 0.1) degrees C, and the cold stress temperature was (4.0 +/- 0.1) degrees C. The rats were treated with different temperatures until 12 h. The abdominal aortic blood was collected with heparin anticoagulation suction tube. Then, the plasma was separated for protein extraction, quantitative, enzymolysis, iTHAQ labeling, scx fractionation and mass spectrometry analysis.</p><p><b>RESULTS</b>Totally, 1085 proteins were identified in the test, 39 differentially expressed proteins were screened, including 29 up-regulated proteins and 10 down-regulated proteins. Three important differentially expressed proteins related to cold stress were screened by bioinfonnatics analysis (Minor histocompatihility protein HA-1, Has-related protein Rap-1b, Integrin beta-1).</p><p><b>CONCLUSION</b>In the experiment, the differentially expressed plasma proteins were successfully screened in cold stress rats. iTRAQ technology provided a good platform to screen protein diaguostic markers on cold stress rats, and laid a good foundation for further. study on animal cold stress mechanism.</p>

Granulosa cells model transfected by ENO1 overexpression adenovirus vector and the effect of ENO1 overexpression on progesterone secretion / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To construct primary cultured granulosa cells model of Zi Gooses tansfected by alpha-enolase (ENO1) overexpression adenovirus vector, and to detect the effect of ENO1 overexpression of granulose cells on progesterone secretion.</p><p><b>METHODS</b>Granulosa cells were infected with Ad-CMV-ENO1 in gradient multiplicity of infection(MOI) levels:100, 250, 350 and 400 pfu/cell. Twenty four hours and 48 h after infection, green fluorescent protein (GDP) was respectively detected by fluorescence inverted microscopy. The effect of ENO1 overexpression of granulose cells on progesterone secretion was detected by the step double antibody sandwich enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The optimal infection rate (100%) was achieved when MOI was 800 pfu/cell,48h after infection. Real time RT-PCR and Western blot showed that the level of mRNA and protein expression ENO1 were increased significantly after infection (P < 0.01); The granulosa cells progesterone secretion of Ad-CMV-ENO1 group increased signigicantly (P < 0.01).</p><p><b>CONCLUSION</b>ENO1 overexpression could make the primary culture follicle granulosa cells in vitro improve progesterone secretion.</p>