| ObjectiveBy observing the effects of tactile stimulation combined with five elements music on behavior,morphology and hippocampal BDNF/Trk B/CREB expression of premature rats,to explore the mechanism of tactile stimulation combined with five elements music in promoting learning and memory ability and hippocampal related protein expression of premature rats,so as to provide theoretical support for rehabilitation treatment of promoting brain development of premature infants.Methods10 clean-grade healthy SD rats and 20 clean-grade healthy SD female rats were selected and fed adaptively for a week after the animals were purcha-sed.The female and male rats were caged according to 2:1,and the pregnant rats were obtained.On the 17 th and 18 th day of pregnancy,the pregnant rats in the LPS group were weighed.The rats in the LPS group were intraperitonea-lly injected with LPS(serotype 055 LPS B5 Magin Sigma Company)350 μg / kg(each 1mg LPS dissolved in 10 ml saline)for 2 days to induce preterm delivery,and the premature animal model was obtained.the offspring born before 22 days of pregnancy were regarded as premature rats.The offspring were randomly divided into 5 groups: normal control group,model control group,five elements music group,tactile stimulation group and combination group with 12 rats in each group.The normal control group did not do any treatment to the normal newborn mice;the model control group did not do any treatment to the postnatal mice;the tactile stimulation group: tactile stimulati-on began 24 hours after birth,15 minutes each time,twice a day,rubbing the back of the offspring repeatedly with a soft brush;the five elements music group: music therapy began 24 hours after birth,15 minutes each time,twice a day.The combined group: tactile stimulation was performed while playing the music of the five elements;all methods were intervened continuously for21 days,and at the corresponding time point at 21 days after intervention,Mirrors water maze was used to detect the learning ability of the offspring in each group,euthanasia was taken,HE staining was used to observe the morphological changes of hippocampal neurons,and WB was used to detect the expression of BDNF,Trk B and CREB in the hippocampus of rats.Result1.The number of pregnant rats,the number of live births and the birth weight of pregnant miceThe results of delivery of pregnant mice were as follows: 18 of the 20 female mice were pregnant successfully,including 3 pregnant mice in the normal group,and 35 full-term mice were delivered in the normal group,and no offspring died.There were 15 pregnant mice in LPS group,of which 2pregnant mice died before delivery,12 pregnant rats delivered early delivery and 1 full-term delivery.In LPS group,96 live-born premature mice,11live-born full-term mice and 16 newborn mice died.The number of live born mice and birth weight in the normal group were significantly higher than those in the model group(P < 0.05).2.Comparison of the results of learning and memory behavior test in different groups of offspring(1)Positioning cruise experiment: with the continuous increase of positioning cruise test days,the escape latency of offspring in each group was also gradually shortened.On the first day of the experiment,the comparison results of escape latency among the three groups were as follows: compared with the normal group,the escape latency of model group,five elements music group and tactile stimulation group was longer,and the difference was statistically significant(P < 0.05).The escape latency of the combined group was prolonged,but there was no significant difference(P > 0.05).Compared with the model group,the escape latency in the five elements music group,the tactile stimulation group and the combination group was shortened(P <0.05),but there was no significant difference between the five elements music group and the tactile stimulation group(P > 0.05).On the second day,the positioning cruising ability of mice in each group was improved,and the escape latency in the normal group,intervention group and model group was shorter than that in the model group(P < 0.05).On the 3rd day,the escape latency in the intervention group was significantly shorter than that in the model group(P < 0.05),and on the 4th day,the escape latency in the combina-tion group was the shortest compared with the model group(P < 0.05).The difference was statistically significant(P < 0.05).(2)Spatial exploration experiment: The results of the times of crossing the platform and the percentage of residence time in the target area in each group were as follows: normal group > combination group > tactile stimulatio-n group > five elements music group > model group.The comparison results of the times of crossing the platform and the percentage of residence time in the target area among the groups were as follows: the number of times crossing the platform and the percentage of stay time in the target area in the model group were significantly lower than those in the normal group(P <0.05).There was no significant difference in the number of times crossing the platform and the percentage of residence time in the target area between the five elements music group and the tactile stimulation group(P > 0.05).There was no significant difference in the times of crossing the platform and the percentage of residence time in the target area between the combination group and the normal group(P > 0.05).The times of crossing the platform and the percentage of residence time in the target area in the combination group were better than those in the five elements music group and the tactile stimulation group(P < 0.05).3.Pathological changes of hippocampal CA1 region in offspring of each groupNormal group: the structure of hippocampal CA1 region was intact and clear,and the neurons were arranged evenly and neatly.Model group: the number of layers of neurons decreased,2-4 layers,arranged sparsely and scattered,the degeneration and necrosis of neurons and the swelling of degenerative neurons could be seen.Five elements music group: the histomor-phology of hippocampal CA1 region had no abnormal changes,the number of neuronal layers was 3-5,and the neurons were arranged neatly and evenly.Tactile stimulation group: the number of neuronal layers in hippocampal CA1 region was more than 3-5 layers,neatly arranged but some areas were sparse,and the cytoplasm was rich and delicate.In the combination group,the number of neurons in hippocampal CA1 region increased significantly,with5-7 layers,neatly arranged,full and delicate cytoplasm,clear nuclear membra-ne and uniform chromatin.4.Relative expression of BDNF,Trk B and CREB proteins in hippocampu-s of newborn mice in each groupThe results of protein expression of BDNF,Trk B and CERB in the hippocampus of newborn mice in each group were as follows: normal group >binding group > tactile stimulation group > five elements music group >model group.The results of comparison among the three groups were as follows: the protein expression of BDNF,Trk B and CREB in the hippocampus of the model group was significantly lower than that of the combination group,but there was no significant difference in the expression of BDNF,Trk B and CREB between the combination group and the normal group,and there was no significant difference in the expression of BDNF,Trk B and CREB between thefive elements music group and the tactile stimulation group.The protein expression of BDNF,Trk B and CREB in the hippocampus of the combined group was better than that of the five elements music group and tactile stimu-lation.Conclusion1.Tactile stimulation and five elements music can significantly enhance the performance of premature mice in Morris water maze test,improve the learning and memory ability of premature mice,and the effect of the combinat-ion of the two is more effective.2.Tactile stimulation and five elements music can improve the neuronal damage in hippocampal CA1 region,and the effect of the combination of tactile stimulation and five elements music is better.3.Tactile stimulation and five elements music can up-regulate the expression of BDNF,Trk B and CREB in hippocampus,and the effect of the combination of the two is better.4.Tactile stimulation and five elements music can improve the learning and memory ability of premature mice,and the mechanism may be related to the up-regulated expression of BDNF,Trk B and CREB. |
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