| Objective: To observe the content of peripheral blood interleukin 6(IL-6),IL-10 and the apoptosis of brain cells,brain tissue Bcl-2,Bax protein expression of Wang Qingren’s different prescriptions of blood stasis on severe craniocerebral injury at acute stage of SD(Sprague-Dawley)rat at different time points(24 h,72 h and 168 h),and to explore the therapeutic effect of severe traumatic brain injury in rats.Methods: 120 SPF(SD)healthy male Sprague-Dawley rats were randomly assigned into normal control group(A),model group(B),Xuefuzhuyu Decoction group(C),Tongqiaohuoxue Decoction group(D)and Buyanghuanwu Decoction group(E).Every group contained with 24 rats.According to the time of after modeling 24 h,72 h and 168 h,the rats were divided into three subgroups,each time point of 8.In addition to the normal control group,the other groups were prepared by modified Feeney’s free fall method to prepare the rat model of severe traumatic brain injury.After modeling 6h,A,B group received saline,while C,D and E groups were gavage administration with liquid extracts of Xuefuzhuyu Decoction,Tongqiaohuoxue Decoction and Buyanghuanwu Decoction.Rats in each group were measured for weight and neurological deficit score(NSS)every day;The materials were collected at three time points on 24 h,72 h,168 h after modeling,The peripheral blood samples and brain tissues were collected.The content of IL-6 and IL-10 in peripheral blood serum were detected by Enzyme linked immunosorbent assay(ELISA);Pathological changes of brain tissue were observed by HE staining;the apoptosis of brain cells tested by TUNEL mothod;the expression of Bcl-2 and Bax in brain tissue were detected by immunohistochemistry.Results:(1)The weight of rats in normal control group increased gradually with time,and reached the highest level at 168h;The weight of rats in model group decreased gradually with time,and decreased to the lowest level at 168 h;After treatment,the weight of rats in each treatment group decreased gradually with the increase of experimental time,but the weight of each treatment group in the model after 72 h,168 h were significantly higher than the model group rats.There were no neurological deficits in normal control rats;The NSS of of model group Xuefuzhuyu Decoction group,Tongqiaohuoxue Decoction treatment group and Buyanghuanwu Decoction treatment group was significantly lower than that of model group anter modeling of 24 h,72 h and 168 h three time points,but there was no significant differences between the three groups at each time point.(2)The normal control group of brain tissue under the microscope structure is complete,and the outline clear,cells arranged in neat rows,while the cell nuclei stained by blue with uniform,there was no tissue edema and necrosis;In the model group,the cell arrangement was disordered and the cells were edematous,besides the local cells were necrotic,and the nuclei were reduced.The brain tissue edema,necrosis and karyopyknosis of Xuefuzhuyu Decoction group,Tongqiaohuoxue Decoction treatment group and Buyanghuanwu Decoction treatment group lower than the model group.(3)The content of IL-6 in peripheral blood serum of model group was significantly increased at 24 h after injury,then down to the lowest level at 168 h,but was still higher than that of normal group.The content of IL-6 in serum with Tongqiaohuoxue Decoction group,Xuefuzhuyu Decoction group and Buyanghuanwu Decoction group was lower than that with model of non drug group at time of 24 h and 72 h.There was no significant difference compared to model group of IL-6 content at 168 h;Serum IL-6 content of Xuefuzhuyu Decoction group and Tongqiaohuouxe Decoction group was significantly lower than Buyanghuanwu Decoction group at 24 h.(4)The content of IL-10 in the peripheral blood serum of the model group and the treatment group was significantly higher than that of the normal control group at 24 h,72 h and 168 h after injury.but there was no statistical difference between the three time points.The content of IL-10 in Xuefuzhuyu Decoction group was the lowest at 24 h,then increased gradually,reached the highest level at 168 h;and it was significantly higher than the model of non medication group at 24 h and 72 h;it was significantly higher than the model group at 168 h.The content of IL-10 in Tongqiaohuoxue Decoction group and Buyanghuanwu Decoction group had no statistical difference compared with non medication group.(5)The brain cells apoptosis appeared immediately after the injury.Compared with normol group,the apoptosis rate of brain cells increased significantly of the model non drug group and the other groups on the three points of the first 24 h,72 h,168 h.The apoptosis rate of model group increasedsignificantly at 24 h,reached the highest level at 72 h,and the apoptosis rate was significantly decreased at 168 h.Compared to the model group,The apoptosis rate of brain cells in each treatment group was significantly lower at 24 h,72 h and 168 h.Compared with Buyanghuanwu Decoction group,the apoptosis rate of brain cells lowered significantly of Xuefuzhuyu Decoction group and Tongqiaohuouxe Decoction group on the two points in 24 h and 72 h.There was no significant difference in TUNEL positive cell rate between Xuefuzhuyu Decoction group and Tongqiaohuouxe Decoction group at three point.(6)The expression of Bcl-2 protein in the model group and the treatment group was significantly higher than that in the normal control group;The expression of Bcl-2 protein in the model group reached a high level after injury,and then began to decline gradually after 24 h.Compared with model group,there was no significant difference on each treatment group at 24 h,and there was increased significantly at 72 h and 168 h,and reached peak at 168 h.Compare to Buyanghuanwu Decoction,the expression of Bcl-2 in Xuefuzhuyu Decoction group and Tongqiaohuoxue Decoction group increased significantly at 72 h.(7)The expression of Bax protein in the model group and the treatment group was significantly higher than that in the normal control group.It is reached the peak at 72 h and then gradually decreased.The expression of Bax protein in rats of each treatment group reached the maximum level at 24 h after injury.The expression decreased gradually with the increase of experimental time.Compared with model group,there was no significant difference among treatment groups of Bax protein’s expression at the time of 24 h,but the expression of the three groups significantly decreased in the 72 h and 168 h.Compared with Tongqiaohuoxue Decoction group and BuyanghuanwuDecoction group,the expression of Bax protein in Xuefuzhuyu Decoction group was significantly decreased at 72 h.Conclusion : Xuefuzhuyu Decoction,Tongqiaohuoxue Decoction and Buyanghuanwu Decoction have a therapeutic effect to nerve injury on acute stage of severe traumatic brain injury in rats.The treatment effect of Xuefuzhuyu Decoction,Tongqiaohuoxue Decoction and Buyanghuanwu Decoction on severe craniocerebral injury may be by reducing the content of IL-6,inhibiting the expression of Bax and increasing the content of IL-10,up-regulating the expression of Bcl-2 to reduce the secondary inflammatory reaction,abate the apoptosis of brain cells and improve the prognosis of severe traumatic brain injury in rats.From the index results,Xuefuzhuyu Decoction has the best effect in three decoctions,followes by Tongqiaohuoxue Decoction,and the last is Buyanghuanwu Decoction. |
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