| Objective:To set up the animal model of hypoxia for experiment and test the effection of the effective components of Valeriana jatamansi Jones on anti hypoxia.Surveying the experimental animals’ behavior and morphology dynamically.Observing and studyingepigenetic changes in order to explore the correlation effect of epigenetic and hypoxia and the effective componentsof Valeriana jatamansi Jones.Methods:1.animal model of hypoxia: 20 SD female rats during the estrous period,according to the ratio of 2:1 and mature SD in April were randomly mated with male rats,after24 hours,respectively,vaginal smear test on each female rats,determined to have a successful mating,the pregnant rats were randomly divided into 4 groups: control group,model group,Valeriana jatamansi Jones treatment group Rhodiola group;on the same day as day 1 of gestation.After hypoxia treatment,the spider incense group,Rhodiola group,the rats in the model group daily in three gas incubator incubator for6 hours,set in 8% oxygen concentration,carbon dioxide concentration of 4%,others was nitrogen.During the period of uninterrupted observation of rat life;placed in the blank group the same humidity and temperature,during the uninterrupted observation of rat model of life;for 7 days,during the modeling,the rats daily observation of the state of mental activity,body weight,food intake,stool,sleep and other general situation.2.Behavioral and histological evaluation of the treatment of hypoxia with spider20 SD rats were randomly divided into 4 groups: blank control group,model group,Rhodiola group and treatment group.Except the blank group in each group according to the method of hypoxia model replication,after successfully modeling,Rhodiola group gavage Nuodikang Capsule CMC-Na solution,Valeriana jatamansi Jones administered Valeriana jatamansi Jones iridiod CMC-Na solution,blank control group and model group were perfused with the same volume of CMC-Na solution,each group’s treatment continue for 7 days.Daily during the treatment observation of rat body weight,food intake,stool,sleep and other general situation,evaluate the disease index;wait until the end of treatment of pregnant rats during production,daily observation of pregnant rats body weight,food intake,stool,sleep and other general observation;F1 rats brain after birth in 7,respectively and 30 day old offspring rats were generally observed brain tissue damage index,grading of brain tissue HE staining;Morris water maze tests were performed on 30 day old offspring rats,the determination of the navigation and air in search ability to infer the generation ability of learning and memory in rats detection,spatial learning and memory function.3.Differential methylation analysis of Me DIP-seq(Methylated DNA Immunoprecipitation Sequencing methylation DNA sequencing)Animal grouping,model replication and treatment as the same.take 30 days of rats’ brain tissue,brain tissue total DNA was analyzed by Me DIP-seq differential methylation,according to the Me DIP-Seq sequence can be high precision Cp G dense hypermethylation regional studies at genomic level,quickly and effectively find methylated regions on the genome,and comparison between different samples of the difference DNA methylation pattern.Results:In the modeling,model group,Valeriana jatamansi Jones group,Rhodiola group rats appear apathetic,lethargy,lazy,his breathing,shortness of breath,diet,body weight decreased,the activity reduced endurance,withered hair messy,that performen as hypoxia.The blank group showed no abnormal manifestation.Valeriana jatamansi Jones treatment group rats increased food intake,compared with the control group rats had no significant difference(P>0.05),body weight has not yet reached the weight of the rats in the blank group at the end of the experiment,but compared with the model group increased significantly(P<0.05),a sensitive response of rats,increased activity,experimental results of the Morris water maze was the difference with the control group,but there were significant differences with the model group;microscopic observation of hippocampus cell damage is not obvious pathological HE staining,compared with model group(P<0.05).Compared with Rhodiola group,Valeriana jatamansi Jones treatment group rats general condition improved obviously,there was no significant difference in body weight food intake(P>0.05),no significant difference between the results of the two Morris water maze test,HE staining,microscope observation showed no significant difference.Differential methylation analysis of Me DIP-seq shows that hypoxia can induce methylation,and Valeriana jatamansi Jones treatment group and Rhodiola treatment group may be different methylation.The intervention mechanism of Valeriana jatamansi Jones against hypoxia may be different from that of Rhodiola.Conclusions:The experimental model is reasonable,and the model is successfully reproduced.Certainly,the effective components of Valeriana jatamansi Jones’ treatment is similar to Salidroside,also can significantly improve the hypoxic rat model to restore mental and physical condition,hypoxia can induce methylation changes.The changes of methylation in gene and signal transduction pathway in the treatment group and the model group were significantly different. |
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