| Background:The difficulty of nerve regeneration after spinal cord injury(SCI)is a worldwide problem.How to improve the local microenvironment after spinal cord injury,delay the progression of secondary injury,and promote neuronal regeneration and repair are the key points of treatment.Hyperplasia of glial scar is considered to be one of the main reasons that nerve axons cannot regenerate and pass through the injured area after injury.As our clinical experience prescription,the traditional Chinese medicine "Spinal Cord Kang" has been found to have analgesic,anti-infflammatory and neurotrophic factor expression promoting effects in previous studies.However,the mechanism of action of"Spinal Cord Kangl in promoting the repair of spinal cord injury is not completely clear.Based on this,we designed clinical research and basic experiments to evaluate the therapeutic effect of "Spinal Cord Kang,on spinal cord injury and explore its mechanism of inhibiting astrocyte activation and promoting spinal cord‘injury repair,so as to provide scientific basis for its treatment of spinal cord injury.Objective:(1)To analyze the curative effect of "Spinal Cord Kang,in promoting the recovery of neurological function in patients with spinal cord injury,and to explore the relevant factors affecting the recovery of neurological function in patients with spinal cord injury;(2)To investigate the role and significance of astrocytes in nerve regeneration by observing the dynamic expression of astrocytes;(3)To evaluate the effect of "Spinal Cord Kang" on promoting the regeneration of nerve function in rats by observing the effect of "Spinal Cord Kang" on the tissue structure of lesion area and muscle strength of rats after spinal cord injury;(1)To observe the effect of "Spinal Cord Kang" on astrocyte morphology,GFAP and CSPG protein expression after spinal cord injury in rats,and to explore its mechanism of promoting nerve regeneration after spinal cord injury.Methods:(4)A total of 68 patients diagnosed as thoracolumbar fracture with incomplete spinal cord injury were collected from the Department of Bone Injury of Wuxi Traditional Chinese Medicine Hospital and divided into control group and observation group according to different treatment schemes.Patients in the control group underwent surgery after admission.The patients were treated with "Spinal Cord Kang" on the basis of the treatment in the control group.The clinical efficacy,sensory and motor scores of the patients in the two groups were observed,and the related factors affecting the prognosis of the patients were analyzed;(5)108 SD rats were divided into three different group.Samples were taken on the 3rd,7th and 14th days after intervention respectively.Immunohistochemistry and Western blot were used to detect the expression changes of GFAP protein in spinal cord tissues of rats in each group,and fluorescence quantitative PCR was used to detect the expression changes of GFAP mRNA;(6)120 SD rats were divided into high dose group,medium dose group,low dose group,prednisone group,sham operation group and model group.The climbing plate score and muscle strength of rats in each group at 3 days,7 days and 14 days were observed,and the morphological and quantitative changes of Nissl bodies,neurons and glial cells in spinal cord tissue were observed;(7)180 SD rats were divided into high dose group,medium dose group,low dose group,prednisone group,sham operation group and model group.Rats in each group were sampled on the 3rd,7th and 14th day.Immunohistochemistry and Western blot were used to detect the expression changes of GFAP and CSPG proteins in spinal cord tissue.Fluorescence quantitative PCR was used to detect the expression changes of GFAP mRNA and CSPG mRNA.Results:(1)Clinical Study on Promoting Nerve Regeneration by Traditional Chinese Medicine "Spinal Cord Kang"68 patients with spinal cord injury were included in the study.The effective rate of treatment in the observation group was 94.3%,while that in the control group was 90.9%.Oral administration of "Spinal Cord Kang" and age are the relevant factors affecting the prognosis of spinal cord injury(P<0.05),while gender is not related to the prognosis(P>0.05);(2)Dynamic expression and significance of astrocytes during nerve regenerationCompared with the sham operation group,the expression of GFAP protein and mRNA in the model group increased after SCI and decreased gradually after reaching the peak at 7d,but it was still higher on 14d than that in the sham operation group,indicating that GFAP,as a representative product of astrocyte activation,was expressed more in the injured spinal cord tissue;(3)Experimental Study on Promoting Nerve Regeneration by Traditional Chinese Medicine "Spinal Cord Kang,After SCI,rats were gavaged with the traditional Chinese medicine "Spinal Cord Kang".The climbing plate data and the neurological function score of both hind limbs of rats were significantly better than that of the model group,indicating that Spinal Cord Kang can promote the neurological function recovery of rats.HE staining showed that.the rats in the model group had obvious spinal cord tissue structure damage,neuronal shrinkage and necrosis,serrere sp:inal cord tissue damage on the 3rd day of modeling,marked aggravation of spinal cord function damage on the 7th day,concurrent glial cell infiltration,aggravated glial cell infiltration on the 14th day,and inflammatory cells began to appear at the same time.Rats in prednisone group and "Spinal Cord Kang,group detected that the destruction degree of spinal cord tissue structure at each node was better than that in the control group;(4)Effect of Chinese Medicine "Spinal Cord Kang" on Expression of GFAP and CSPG in Astrocytes of Rats with Spinal Cord InjuryIumunohistochemistry,Western blot and fluorescence quantitative PCR showed that the expressions of GFAP,CSPG protein and mRNA in the model group after SCI were significantly higher than those in prednisone group and spinal cord rehabilitation group(p<0.05).The expressions of GFAP,CSPG protein and mRNA in prednisone group,spinal cord kang high,medium and low dose groups showed dynamic changes on the 3rd,7th and 14th day.After successful modeling,the expression increased on the 3rd day,peaked on the 7th day,and then gradually decreased.Conclusion:(1)"Spinal Cord Kang" can promote neurological rehabilitation of patients with spinal cord injury;(2)The expression level of GFAP and CSPG protein in spinal cord tissue is negatively correlated with the degree of neurological function recovery after spinal cord injury,indicating that "Spinal Cord Kang" promotes the repair of spinal cord injury by inhibiting the expression of GFAP and CSPG protein,which are representative products of astrocyte activation. |
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