Transcriptome Sequencing On The Effect Of Gene Expression Of Acute Spinal Cord Injury In Rats With Ligustrazine

Objective: In order to provide theoretical basis for the mechanism of ligustrazine in the treatment of spinal cord injury,the differentially expressed genes(DEG)of acute spinal cord injury(SCI)in rats after Ligustrazine intervention were investigated based on transcriptome sequencing.Methods: Thirty SPF healthy male SD rats of Six-eight weeks old,weighing 200-230 g,were randomly divided into three groups: 6 rats in the sham-operated group(group A / A14d),model group(group B,including 6rats in group B7 d and 6 rats in group B14d)has 12 rats,12 rats in the Ligustrazine intervention group(Group C,including 6 rats in group C7 d and6 rats in C14 d group);In group A,only spinal cord tissue was exposed and sutured,the acute spinal cord injury model was established in group B and group C.After successful modeling,group C was intraperitoneally injected with Ligustrazine(100 mg / kg,10 mg / ml,prepared with normal saline),and group A and group B were received intraperitoneal injection of normal saline.BBB score was performed before modeling,7 days(d)and 14 days after modeling in each group.In B7 d group and C7 d group,samples were collected and made at 7 days after successful modeling;and specimens were collected and made at 14 days after successful modeling in Group A,B14 d group and C14 d group.In each group,3 specimens were randomly stained with HE and Nissl staining,followed by pathomorphological observation,and 3 specimens were transcriptome sequenced,screened and compared for gene differentialexpression,and their gene ontology(GO)and Kyoto Encyclopedia of Gene and Genome Database(KEGG)signal pathway enrichment analysis were performed.Results: 1.BBB score suggested that there was no spinal cord injury in the sham operation group;compared with the model group,the BBB score was significantly higher in the Ligustrazine intervention group(P < 0.05).2.The results of HE staining and Nissl staining showed that: compared with group A,spinal cord tissue injury could be seen in both group B and group C.At 7d and 14 d after modeling,the recovery degree of spinal cord injury in group C was better than that in group B,and the number of spinal cord neurons and Nissl bodies was significantly more than that in group B.3.High-throughput transcriptome sequencing showed that 70 DEGs of B7d-vs-C7 d,including 42 up-expressed DEGs and 28 down-expressed DEGs;66 DEGs of B14d-vs-C14 d,including 31 up-expressed DEGs and 35down-expressed DEGs;886 DEGs of A-vs-B7 d,including 702 up-expressed and 184 down-expressed;1404 DEGs of A-vs-B14 d,including 921up-expressed and 483 down-expressed.4.GO enrichment analysis results showed that the vast majority of the groups were enriched in biological processes(BP);B7d-vs-C7 d was enriched in BP300 items,cell component(CC)34 items and molecular function(MF)86items,mainly involving inflammation,immune response,neuronal ion channel aggregation,hemostasis,pain,synapse reconstruction,axonal sensation,myelination and so on;B14d-vs-C14 d was enriched in BP 413 items,CC 13 items and MF 59 items were mainly related to immune regulation,catecholamine metabolism,cytokine-cytokine receptor interaction,neuroactive ligand-receptor interaction,GABAergic synapse,dopaminergic synapse,oxidative stress injury,etc.A-vs-B7 d was enriched in BP 1796 items,CC 102 items and MF 286 items,A-vs-B14 d was enriched in BP 2151 articles,CC 174 articles and MF 274 articles.5.KEGG enrichment analysis results showed that five pathways were significantly enriched in B7d-vs-C7 d,including Cell adhesion molecules(CAMs),complement and coagulation cascades,Hedgehog signaling pathway,etc.13 pathways were significantly enriched in B14d-vs-C14 d,including cytokine-cytokine receptor interaction,retrograde endocannabinoid signaling,neuroactive ligand-receptor interaction,PPAR signaling pathway,GABAergic synapse,Fox O Signaling pathway,dopaminergic synapses,etc.86 pathways were significantly enriched in A-vs-B7 d and 95 pathways were significantly enriched in A-vs-B14 d.Conclusion: Ligustrazine can protect the cell apoptosis and nerve injury of spinal cord injury through inflammatory response,immune response,neuron ion channel aggregation,oxidative stress injury,hemostasis,synaptic reconstruction,neuropathic pain,GABAergic synapse,hedgehog signaling pathway,PPAR signaling pathway,TGF-? signaling pathway,ERBB2-ERBB 3 signaling pathway,Wnt signaling pathway,BMP signaling pathway and other processes.