The Mechanism Of MiR-200c In Cerebral Protection Of Deep Hypothermic Circulatory Arrest

Purpose This experiment in 350 ～ 400 g in weight between the healthy adult male SD rats as experimental object,establish deep cryogenic stop cycle model in rats,blocking bilateral carotid artery,simulation of the deep cold stop circulation ischemia hypoxia induced brain injury,Real time PCR method and FISH techniques were used to detect miR-200-c expression in hippocampus of rats,respectively measuring stop at deep low temperature circulation cerebral ischemia of rats induced by hypoxia brain injury after 6hours,24 hours,48 hours of miR-200-c expression quantity changes,and the negative control group,the differences of expression Observation of miR-200-c expression decreased trend and brain injury degree of consistency,using the in situ hybridization technique for gene amplification,the fluorescence microscope miR-200-c model in rats in DHCA the expression and distribution of hippocampal CA1 area,and the negative control group no significant brain damage of immunofluorescence images compared brain tissue,the preliminary judgment of miR-200-c protective effect of nerve cells,for the prevention and treatment of brain injury postoperative opened up a new direction.Methods The SD rats used in this experiment were purchased from a biological company and were in accordance with the regulations on the management of experimental animals.1.Ten healthy adult male SD rats with body weight between 350 and 400 g were randomly divided into two groups: DHCA group(n=5)and control group(n=5).The rats in control group were given anesthesia and hypothermia treatment only,and bilateral common carotid artery and femoral artery bloodletting was not blocked.DHCA group was given after anesthesia and deep low temperature around(21 ℃)connection heparinization function of biological systems,from the femoral artery blood step-down and clip carotid artery to simulate the brain injury induced by cerebral ischemia hypoxia model,and connect the ventilator,ecg monitoring,the monitoring of heart rate,blood pressure,body temperature,remove tiny carotid artery after 60 min clip,and femoral artery into the blood,thawing,maintain body temperature and smooth breathing,heart rate after the big mouse,back in the cage raising survival.2.spine was put to death in the rat brain tissue after hippocampal CAl area,formaldehyde solution fixed 2 days later,the frozen section,using the method of dye Hairpin-it miRs quantitative calibration and u6 qRT-PCR kit to detect the expression level of miR-200-c,after using in situ hybridization technique of the target gene amplification in fluorescence microscope miR-200-c model in rats in DHCA the expression and distribution of hippocampal CA1 area.Results1.The cerebral ischemia injury model of deep hypothermic circulatory arrest in rats was successfully established;2.The brain tissue of hippocampal CA1 region of rats in the deep hypothalmic circulation arrest model group was taken,and the miRNAs in the brain tissue were successfully extracted with the miRNAs rapid extraction kit;3.The miRs quantitation and U6 calibrated qRT-PCR kit were used by the dye method Hairpin-it to accurately detect the expression level of miR-200c;4.The expression and distribution of miR-200c in the hippocampal CA1 region of DHCA rat model were observed by in situ hybridization and fluorescence microscopy,and its fluorescence images were obtained successfully.5.It was found by comparison that the miR-200c content in the brain tissues of the rats with DHCA ischemia injury was lower than that of the control group.The SPSS23.0 software was used to perform T test on the examination results of the control group,and the analysis showed a statistical difference.Conclusion1.The expression level of miR-200c in brain cells will change with cerebral ischemia-hypoxia-induced injury.When brain tissue is damaged,the expression level of miR-200c will decrease,but there is no such trend in the absence of cerebral ischemia injury.2.The expression of miR-200c in the brain tissues of the negative control group rats without obvious brain injury showed no significant change,while the expression of miR-200c in the brain tissues of the DHCA group rats with brain injury showed a trend of decrease,suggesting that miR-200c may be a protective factor of brain injury.