| Purpose:In this experimental study,by observing the changes of Xiaoshuanjiannao Granules on nerve injury,cerebral infarction area,brain edema,pathology,mitochondrial membrane potential and the differences of gene and protein expression of NLRP3 and caspase-1 in cerebral infarction rat models,the mechanism of Xiaoshuanjiannao Granules in treating cerebral infarction is discussed,so as to provide experimental and theoretical basis for clinical treatment of cerebral infarction in the future,to improve and promote the development of cerebral infarction treatment.Materials and methods: Forty SPF grade SD rats,aged 6-8 weeks,all male,and weighing 250±20g,were randomly divided into sham operation group,model group,experimental group and control group,with 10 rats in each group.The model of cerebral infarction in rats was established by thread embolism method.After successful modeling,rats in the sham operation group and model group were given intragastric administration of normal saline every day,rats in the experimental group were given intragastric administration of Xiaoshuanjiannao Granules,and rats in the control group were given intragastric administration of Xuesaitong tablets.The rats in each group were tested for neurological function with Zea Longa score on the 1st and 14 th day after operation to determine whether the establishment of the model was successful and the condition of nerve function injury.After 14 days of continuous gastric perfusion,HE staining was used to observe the pathological changes of brain tissue.,TTC staining was used to detect the area of cerebral infarction,brain water content was calculated to evaluate the degree of cerebral edema,mitochondrial membrane potential was detected,and the expressions of Caspase-1 and NLRP3 genes and proteins were detected by real-time fluorescence quantitative PCR and Western Blot.Results: 1 Neurological function test resultsAfter model surgery,the results of Longa score showed that the score of rats in the sham operation group were lower than that of in the model group,the experimental group and the control group on the 1st after modeling,with statistical difference(P< 0.01),indicating that the cerebral infarction model established in this experiment was replicated successfully.When 14 days after operation,compared with the model group,the Longa score of rats in the experimental group and the control group decreased significantly(P< 0.01);compared with the control group,the Longa score of rats in the experimental group had no difference(P> 0.05).2 HE staining resultsIn the sham operation group,neurons were normal in morphology,complete in structure,clear in nucleolus and without obvious pathological changes;in the model group,neuron cells swelling,cell vacuolization,cnucleus shrinkage and disappearance,and the number of neuron cells significantly decreased;in the experimental group,a small number of neuron cells were swelled,pyknosis of nuclei,and the number of neuron cells significantly increased compared with the model group;in the control group,a small number of neuronal damage were occasionally seen,the whole neurons and tissue morphology were in good shape,the number of neuron cells increased significantly compared with the model group.3 Results of TTC staining in detecting cerebral infarction volumeIn the sham operated group,there was no infarct in bilateral brain tissues,and all brain tissues were red stained;compared with the sham operated group,the rats in the experimental group,the model group and the control group showed obvious cerebral infarction(P< 0.01);compared with the model group,the area of cerebral infarction of rats in the experimental group and the control group decreased significantly(P< 0.01);compared with the control group,there was no significant difference in cerebral infarction area between the experimental group and the control group(P>0.05).4 Brain water content measurement resultsCompared with the sham operation group,the brain water content of rats in the model group increased significantly(P<0.01),while that of rats in the experimental group and the control group increased(P< 0.05);compared with the model group,the brain water content of rats in the experimental group and the control group decreased(P<0.05).Compared with the control group,there was no significant difference in the brain water content of rats in the experimental group(P>0.05).5 Test results of mitochondrial membrane potentialCompared with the sham operation group,the fluorescence intensity of rats in the model group and the experimental group decreased significantly(P<0.01),while that of rats in the control group decreased(P<0.05);compared with the model group,the fluorescence intensity of rats in the experimental group increased(P<0.05),while that of rats in the control group increased significantly(P<0.01);compared with the control group,the fluorescence intensity of rats in the experimental group had no significant difference(P>0.05).6 Test results of real time fluorescent quantitative PCR 6.1 Results of caspase-1 detected by real-time fluorescence quantitative PCRThe melting curve of caspase-1 shows a single peak curve,indicating that it can be amplified specifically.Expression results of caspase-1 in rats brain tissue in each group: compared with the sham operated group,caspase-1 expression in brain tissue of rats in the model group and the experimental group increased significantly(P<0.01),while caspase-1 expression in brain tissue of rats in the control group increased(P<0.05);compared with the model group,caspase-1 expression in brain tissue of rats in the experimental group and the control group decreased significantly(P<0.01),compared with the control group,caspase-1 expression in brain tissue of rats in the experimental group increased,with statistical difference(P<0.05).6.2 Results of NLRP3 detected by real-time fluorescence quantitative PCRThe melting curve of NLRP3 shows a single peak curve,indicating that it can be amplified specifically.Expression results of NLRP3 in rats brain tissue in each group: compared with the sham operated group,NLRP3 expression in brain tissue of rats in the model group,the experimental group and the control group increased significantly(P<0.01);compared with the model group,NLRP3 expression in brain tissue of rats in the experimental group and the control group decreased significantly(P<0.01);compared with the control group,NLRP3 expression in brain tissue of rats in the experimental group increased,with statistical difference(P<0.05).7 Western blot test results 7.1 Results of Caspase-1 detected by Western blotCompared with the sham operation group,caspase-1 expression in brain tissue of rats in the model group and the experimental group increased significantly(P< 0.01),while the caspase-1 expression in brain tissue of rats in the control group increased(P< 0.05);compared with the model group,caspase-1 expression in brain tissue of rats in the experimental group and the control group decreased significantly(P< 0.01);compared with the control group,caspase-1 expression in brain tissue of rats in the experimental group increased,with statistical difference(P< 0.05).7.2 Results of NLRP3 detected by Western blotCompared with the sham operation group,NLRP3 expression in brain tissue of rats in the model group and the experimental group increased significantly(P<0.01),while that in the control group increased(P<0.05);compared with the model group,NLRP3 expression in brain tissue of rats in the experimental group and the control group decreased significantly(P<0.01);compared with the control group,NLRP3 expression in brain tissue of rats in the experimental group increased,with statistical difference(P< 0.05).Conclusion: 1.In this experiment,the model of cerebral infarction in rats was successfully established and used in the experiment by using the method of thread embolism.2.Xiaoshuanjiannao Granules can obviously improve the nerve injury behavior of rats with cerebral infarction,inhibit the occurrence of cerebral edema,effectively increase the mitochondrial membrane potential of brain tissue,and play a role in reducing the area of cerebral infarction.3.Xiaoshuanjiannao Granules can protect the damaged neuron cells and improve the pathological damage morphology of rats with cerebral infarction.4.Xiaoshuanjiannao Granules can effectively inhibit the inflammatory response,down regulate the gene and protein expression of caspase-1 and NLRP3 inflammatory factors.5.By improving the pathomorphology,brain edema,mitochondrial membrane potential of brain tissue and the inhibition of caspase-1 and NLRP3 inflammatory factors expression in rats with cerebral infarction,so as to reduce the area of cerebral infarction,which may be one of the mechanisms of Xiaoshuanjiannao Granules in protecting brain tissue and nerves in rats model with cerebral infarction. |
PDF Full Text Request