Effects Of Xuefu Zhuyu Decoction And Exogenous Thyroxine On Nerve Regeneration And Repair In Rats With Severe Craniocerebral Injury

Objective The effects of Xuefu Zhuyu Decoction（XFZY Decoction）on the expression of BDNF and NGF protein in the hippocampus of rats with severe craniocerebral injury were observed to explore the effect of its regenerative repair on the rats with severe head injury.Method A total of 90 SPF grade healthy male SD rats were randomly divided into 5 groups:normal control group,model non drug group,XFZY low dose group,XFZY medium dose group and XFZY high dose group,18 rats in each group.The rats were divided into three subgroups according to the model at 1^st day,3^rd day and 7^th day,and 6 rats were at each time point.In addition to the normal control group,the other groups were prepared by modified Feeney’s free falling body method to prepare the rat model of severe head injury.Normal control group and model group were given normal saline after 6h,In the low,middle and high dose groups of XFZY Decoction,the rats were given intragastric administration of XFZY Decoction at the corresponding concentration,once a day,3ml/times,for 7 consecutive days.6 rats were taken for neurological deficit score at the 1^st day,3^rd day,7^th day,and then anesthetized rats were collected for peripheral blood and brain tissue samples.The pathological changes of brain tissue were observed,and the expression of BDNF and NGF protein in hippocampus of rats were detected.Results（1）The score of nerve function defect in the rats with severe craniocerebral injury was in a high level.With the prolonged time,the score decreased gradually and the lowest on the 7^th day（P<0.05）.After the low dose of XFZY Decoction,the score of nerve function defect of rats decreased significantly in 7^th day（P<0.05）.The score of nerve function defect in the middle dose group decreased significantly（P<0.05）on 3^rd day and 7^th day,and there was no significant change in the high dose group of thyroxine XFZY Decoction.（2）The brain tissue structure of the normal control group was complete without pathological changes.The model non medication group could see the cerebral interstitial hemorrhage,inflammatory cell infiltration,nuclear condensation and so on.With the time prolonging,the pathological changes in the low,middle and high dose groups of XFZY Decoction were lighter than those of the model group at the three time points.（3）The expression of BDNF protein in the hippocampus of normal control rats was very seldom,and the expression of two proteins in the hippocampus of the rats in the low,middle and high dose groups of the model non medication group and XFZY Decoction increased significantly at the three time points after the injury（P<0.05）.Compared with the model non medication group,the low dose and middle dose group of XFZY Decoction were all increased at 7^th day,while the high-dose group did not change significantly at three time points.（4）Compared with the normal control group,the expression of NGF in the hippocampus of the other four groups was significantly increased at three time points after injury（P<0.05）.Compared with the model group,the expression of NGF in the low dose group and middle dose group increased significantly on the third day（P<0.05）,and lasted for 7^th day.There was no significant change in the high dose group.Conclusion XFZY Decoction can promote the repair of nerve tissue in rats with severe craniocerebral injury by promoting secretion of BDNF and NGF protein.Objective To observe the effect of thyroxine on the expression of Wnt3 a m RNA,β-catenin m RNA and the expression of BDNF,NGF and S100 B protein in hippocampus of rats with severe craniocerebral injury,and to explore the effect of thyroxine on nerve regeneration and repair in rats with severe craniocerebral injury.Method A total of 108 SPF grade healthy male SD rats were randomly divided into 6groups:normal control group,normal intervention group,model non drug group,thyroxine low dose group,thyroxine medium dose group and thyroxine high dose group,18 rats in each group.The rats were divided into three subgroups according to the model at 1st day,3rd day and 7th day,and 6 rats were at each time point.In addition to the normal control group and the normal intervention group,the other groups were prepared by modified Feeney’s free falling body method to prepare the rat model of severe head injury.Normal control group and model group were given normal saline after 6h,the normal control group and model treatment group were given saline normal thyroxine intervention group,low dose group,middle dose group and the thyroid hormone thyroxine in high dose group given thyroxine orally administered with 2ml once daily for 7 consecutive days,and 6 rats were taken for neurological deficit score at the 1st day,3rd day,7th day,and then anesthetized rats were collected for peripheral blood and brain tissue samples.ELISA method was used to detect the level of S100 B protein in peripheral blood.HE staining was used to observe the pathological changes of brain tissue.RT-PCR method was used to detect the relative expression of Wnt3 a and β-catenin m RNA in hippocampus,and the expression of BDNF and NGF protein in hippocampus was detected by immunohistochemistry.Results（1）There was no nerve function defect in normal control group and normal control group.After the model,the score of the nerve function defect in the rats with severe craniocerebral injury was significantly increased.With the time prolonging,the score of the neural function decreased gradually and decreased to the lowest in the 7th day（P<0.05）.After the thyroxine was given,the degree of nerve function defect was obviously improved in each group.In the 3rd day and 7th day,the score of the middle dose and the high dose of thyroxine decreased significantly（P<0.05）.However,there was no significant change in the low dose of thyroxine.（2）The content of S100 B protein in rat serum was significantly increased at 1st day after severe craniocerebral injury（P<0.05）,and lasted for 7th day.The levels of S100 B in the thyroid hormone and high dose group decreased significantly at 3th day（P<0.05）,but there was no significant change in the low dose thyroxine group.（3）To observe the normal control group and normal intervention group rat brain structure without pathological changes under light microscope;model of non visible brain hemorrhage clearance treatment group rats,infiltration of inflammatory cells,nuclear pyknosis,and the damage aggravated with the prolongation of time,thyroxine,low dose group,middle dose group and high dose group three period pathological changes were all lighter than that of model group.（4）Compared with the normal control group,model group,the expression of non thyroid hormone low dose group,middle dose group and high dose group rats hippocampus Wnt3 a m RNA after injury 1st day,3rd day and 7th day were significantly increased（P<0.05）;normal intervention group had no obvious change.Compared with the model group,the expression of Wnt3 a m RNA in thyroid hormone group increased significantly on the 7th day.In the middle dose group and the high dose group,the expression of P increased significantly in 3rd day and 7th day（P<0.05）,but there was no significant difference in the degree of increase between the two groups.The expression of Wnt3 a m RNA in model non drug group and thyroxine low dose group was not significantly changed on the 1st and the 3rd day,and increased significantly on the 7th day（P<0.05）.The thyroid hormone in the middle dose group and the high-dose group increased significantly with time,and increased to the peak value on seventh days（P<0.05）.（5）Compared with normal control rats,there were no obvious change of β-catenin m RNAin normal intervention group,while the expression of non medication group,and low dose group,middle dose group and high dose group of thyroxine in hippocampus of rats were significantly up-regulated at three time points（P<0.05）.Compared with the model thyroxine medication group,low dose group increased significantly in 7th day,middle dose group and high dose group significantly increased at three time points（P<0.05）.The expression of β-catenin m RNA in normal control group,normal intervention group and model non drug group did not change with time.The expression of β-catenin m RAN in low dose thyroxine group increased significantly at 7th day（P<0.05）,while the medium dose group and the high dose group increased gradually with time,and reached the peak value at 7th day（P<0.05）,bue there was no significant difference between the two groups.（6）There were little expression of BDNF in normal control group and normal group in hippocampus of rats,while the remaining four groups of BDNF protein in 1st day,3rd day and 7th day were significantly increased after injury（P<0.05）.Compared with the model group,there were not changed significantly in low dose thyroxine group at 1st day and 3rd day and 7th day,while the middle dose group and high dose group rats at three time points were significantly increased.Compared with thyroxine in low dose group,the middle dose group and high dose group rats were significantly increased at three time points（P<0.05）.In the same group,BDNF in normal control group and normal intervention group did not change significantly at three time points.The non drug group reached the peak on the 3rd day,and decreased in 7th day,but it was still higher than that in the normal control group（P<0.05）.The middle dose and high dose thyroxine group reached peak value on the 3st and decreased significantly in 7th day（P<0.05）.（7）There was a little NGF protein expression in hippocampus cells of normal control group and normal intervention group.Compared with the normal control group,the expression of NGF protein in model non medication group,the low dose of thyroxine dose group,the middle dose group and the high dose group all increased significantly at three time points（P<0.05）.Compared with the model group,the rats in the low-dose thyroxine group did not change significantly.Compared with model non drug group and low dose thyroxine group,the expression of NGF in middle dose group and high dose group increased significantly at three time points（P<0.05）.In the same group,There was no significantly change of NGF in normal control group,normal intervention group and low-dose thyroxine group at three time points,but the other three groups reached the peak on 3st days,while decreased at 7th days,but still significantly higher than the normal control group（P<0.05）.Conclusion Exogenous thyroxine has no effect on the Wnt/β-catenin signaling pathway in normal rats,while in the case of craniocerebral injury,exogenous thyroxine can obviously promote the expression of m RNA in this signaling pathway and the secretion of BDNF and NGF,also inhibit the level of S100 B protein.