| Objective: The main content of this study is the expression of tissue inhibitor of metalloproteinase-1(TIMP-1)and matrix metalloproteinase-9(MMP-9)in epileptic patients,acute epileptic rats and chronic epileptic rats,so as to explore the influence of imbalance of MMP-9/TIMP-1 on brain injury caused by epilepsy and explore the methods to improve brain injury in epileptic patients.Methods:(1)Animal experiment: Rats with appropriate weight were selected as the experimental objects and randomly divided into control group(n=12)and PILO-induced epilepsy group(n=28).Pilocarpine-induced epilepsy group rats used pilocarpine to induce tonic convulsion status(SE).Rats in the control group were injected with normal saline.Pilocarpine group was randomly divided into two subgroups(n=14): acute group(E6 h)and chronic group(E90d),with 14 epileptic rats in each subgroup.Observe the behavior of rats,according to Racine grading standard,the success of modeling is that the attack reaches grade 4 or above.In this study,Western Blot,real-time quantitative PCR and immunohistochemistry were used to observe the expression levels of TIMP-1 and MMP-9 in brain tissue of epileptic group and control group.(2)Clinical study: This study was divided into epilepsy group(n=10)and control group(n=3).The epilepsy group: brain tissue samples from patients who underwent temporal lobe resection in the Affiliated Hospital of Jining Medical College due to intractable epilepsy,and the control group: brain tissue samples from patients with craniocerebral trauma;Western Blot,fluorescence quantitative PCR and immunohistochemistry were used to detect the expression differences of TIMP-1 and MMP-9 between the two groups.Results:(1)The success of epilepsy model was 11 rats in acute epilepsy group and 9 rats in chronic epilepsy group,and there was no death in saline injection group.(2)Western Blot showed that the expressions of TIMP-1(p < 0.001,n=11)and MMP-9(p< 0.001,n = 11)were increased in the acute epileptic rat model(Figure 1);The expression of MMP-9 was increased in the chronic epilepsy model(p < 0.001,n=9),but the expression of TIMP-1 was decreased compared with the control group(p< 0.001,n = 9)(Figure 2).In the brain tissue of epileptic patients,the expression of MMP-9 increased(p< 0.001,n=10)and the expression of TIMP-1 decreased(p< 0.001,n=10)(Figure 3).(3)The results of real-time quantitative PCR indicated that the expressions of TIMP-1(p<0.001,n=11)and MMP-9(p< 0.001,n = 11)were increased in the acute epileptic rat model.In the rat model of chronic epilepsy,the expression of MMP-9((p< 0.01,n = 9)increased,while the expression of TIMP-1((p<0.01,n = 9)decreased.In the brain tissue of patients with chronic epilepsy,the expression of MMP-9(p< 0.001,n=10)increased,while the expression of TIMP-1(p<0.01,n=10)decreased.(4)Immunohistochemical results showed that compared with the control group,the protein levels of TIMP-1(Figure 7b)and MMP9(Figure 7d)were significantly increased in acute epilepsy group,while in chronic epilepsy group,the expression of MMP9 was enhanced(Figure 8b),but the expression of TIMP-1 was decreased(Figure 8d).Compared with the control group,the expression of MMP-9(fig.6d)and TIMP-1(fig.6b)in the patients with epilepsy increased in immunohistochemistry,but decreased.Conclusion:(1)In this study,the expressions of TIMP-1 and MMP-9were verified in the brain tissues of rats with acute and chronic epilepsy and patients with epilepsy for the first time.The results showed that the expression levels of TIMP-1 and MMP-9 in the brain tissues of rats with acute epilepsy increased.In the model group of chronic epilepsy rats,the expression of TIMP-1 decreased,while the expression of MMP-9continued to increase.In the brain tissue of epileptic patients,the expression of TIMP-1 decreased and the expression of MMP-9 increased.(2)Consistent with previous research results,in the early stage of epileptic seizure,TIMP-1 expression increased,and MMP-9 was inhibited,thus protecting brain tissue.(3)MMP-9 may be an objective biological index to judge brain injury.(4)TIMP-1 may become a potential antiepileptic drug,and it is believed that with the continuous progress of research,a greater breakthrough will be made. |
