Based On SHH Signaling Pathway To Explore The Mechanism Of Tongdu Huoxue Decoction In The Treatment Of Acute Spinal Cord Injury

Objects:By observing the effect of Tongdu Huoxue Decoction on acute spinal cord injury in rats,the relationship between its role and SHH signaling pathway-mediated inflammation was clarified,and the mechanism of Tongdu Huoxue Decoction on acute spinal cord injury was explored.Methods:Fifty-six SPF SD rats(male and female)were divided into 8 sham operation groups,24 model groups,and 24 Tongdu Huoxue Decoction groups after laminectomy.After the different time,it is divided into three subgroups: 1d,3d,and 7d).In the sham operation group,only the laminectomy was performed without striking the spinal cord.The remaining two groups used a modified Allen 's blower to attack the spinal cord to prepare an acute spinal cord injury model.Rats in the Tongdu Huoxue Decoction group were given gavage of Tongdu Huoxue Decoction every day from the 1st day after surgery,and the remaining two groups of rats were given the same amount of normal saline.Rats in the sham-operated group were scored after intervention at 1d,3d and 7d and scored after 7d;the remaining two groups of rats were scored at intervention 1d,3d and 7d respectively and scored after scoring.BBB and Reuter scores were used to evaluate the sensory motor function of hindlimbs in each group;HE and Nissl staining were used to observe the morphology and structure of the spinal cord and neurons in each group;Elisa test was used to detect IL-1? and TNF-? in the spinal cord tissue of each group.Western Blot detection method to detect the expression of SHH,GLi1 protein in the spinal cord tissue of each group of rats;qPCR detection of IL-1?,TNF-?,SHH,GLi1 mRNA expression in the spinal cord tissue of each group of rats.Results:1.BBB and Reuter scores: Compared with the sham operation group,the BBB scores of the other two groups were significantly lower(P<0.01),and the Reuter score was significantly higher(P<0.01).Compared with the model group,The BBB score of rats in the group of Tongdu Huoxue Decoction increased significantly after 3 days(P <0.01),and the Reuter score decreased significantly after 7 days(P <0.01).2.HE and Nissl staining: In the sham-operated group,the spinal cord morphology was intact,the gray-white matter was clearly demarcated and the gray matter was butterfly-shaped,the Nissl body was full,and it was tabby-shaped,and there was no nuclear collapse or hollow.The morphology and structure of the spinal cord changed due to the impact of the spinal cord,but compared with the model group,the edema and inflammatory reaction in the spinal cord tissue of the rats in the Tongdu Huoxue Decoction group were less light after injury,and the Nissl body was relatively full and clear,and the nucleus There are relatively few neuronal cells that collapse,dissolve,and disappear.3.Elisa: Compared with the levels of IL-1? and TNF-? in the spinal cord of rats in the sham operation group,the levels of IL-1? and TNF-? in the other two groups were significantly increased(P<0.01);compared with the model group.The levels of IL-1? and TNF-? in Tongdu Huoxue Decoction group were significantly decreased(P<0.05 or P<0.01).4.Western Blot: Compared with the expression levels of SHH and GLi1 in the spinalcord tissue of the sham-operated group,the expression levels of SHH and GLi1 in the remaining two groups were significantly increased(P <0.05);compared with the model group,the SHH and GLi1 in the Tongdu Huoxue Decoction group The expression level was significantly increased(P <0.05 or P <0.01).5.qPCR: Compared with the expression of IL-1?,TNF-?,SHH,and GLi1 mRNA in the spinal cord tissue of the sham-operated group,the expression of IL-1?,TNF-?,SHH,and GLi1 mRNA in the remaining two groups was significantly increased(P <0.01);Compared with the model group,the expression levels of IL-1? and TNF-?mRNA in Tongdu huoxue Decoction group were significantly reduced(P <0.05 or P <0.01),and the expression levels of SHH and GLi1 mRNA were significantly increased(P <0.05 or P <0.01)Conclusions:1.Tongdu Huoxue Decoction can promote the recovery of sensory motor function of hind limbs of rats after acute spinal cord injury.2.Tongdu Huoxue Decoction can repair the acute spinal cord injury by promoting the SHH signaling pathway after acute spinal cord injury,inhibiting the inflammatory response after injury.