Effect Of Xingnaojing On Autophagy Of Hippocampus Neurons In SAH Rats

Objectives Objective To investigate the effect of Xingnaojing injection on autophagy in hippocampus of rats with subarachnoid Hemorrhage(SAH).Methods Part I of the experiment,36 healthy clean male Sprague-Dawley rats(350±30g)were selected and serially numbered.The rats were equally divided into sham operation group(Sham group)and subarachnoid hemorrhage group(SAH Group),SAH+Xingnaojing injection treatment group(Drug group)three groups.The Sham group only inserted the core into the internal carotid artery without puncturing the blood vessel.The SAH model reference was prepared by the internal carotid artery puncture method.The Drug group was given intraperitoneal injection of Xingnaojing injection after successful modeling.24 hours after the success is the time node.The shuttle box test was used to observe the time of escape response(ERT)and the numeber of escape response(ERN)in each group of rats.The changes of learning and memory behaviors of rats in each group were compared.HE staining was used to observe the morphological structure of nerve cells in hippocampus of each model group.Immunohistochemical staining and Western Blot assay were used to analyze the expression of Beclin-1 and LC3-II protein of nerve cells in rat hippocampus.Part II of the experiment,40 healthy clean male Sprague-Dawley rats(350±30g)were selected and serially numbered and the rats were aliquoted into 3-MA(3-methyl adenine)group,3-MA+Xingnaojing injection group,Rapa(rapamycin)group,Rapa+Xingnaojing injection group were divided into four groups.The preparation method of SAH model was the same as experiment 1.Each group was given intraperitoneal injection of 3-MA(3-methyl adenine),3-MA+Xingnaojing injection,Rapa,Rapa+Xingnaojing injection respectively after successful modeling.Intervention,select the time node 24 hours after successful modeling.Western Blot assay was used to analyze the expression of Beclin-1 and LC3-II protein of nerve cells in rat hippocampus.Results Part I of the experiment : 1 The shuttle box test:Compared with the sham group,the time of escape response(ERT)was prolonged(P<0.05)and the numeber of escape response(ERN)was decreased(P<0.05)in the SAH group.Compared with the SAH group,the time of escape response(ERT)was decreased(P<0.05)and the numeber of escape response(ERN)was increased(P<0.05)in the drug group.2 HE staining results: Most of the neurons in the hippocampal CA1 area of the Sham group were normal.Compared with the sham group,the numbers of normal nerve cells in hippocampus CA1 area were reduced(P<0.05)in the SAH group.Compared with the SAH group,the numbers of normal nerve cells in hippocampal CA1 area were increased(P<0.05)in the drug group.3Immunohistochemistry and western Blot results: Immunohistochemistry: Compared with the sham group,the numbers of Beclin-1 and LC3-?positive cells were increased(P<0.05)in the SAH group.Compared with the SAH group,the numbers of Beclin-1 and LC3-?positive cells were increased(P<0.05)in the drug group.Western Blot results:Compared with the sham group,the expression level of Beclin-1 protein and the ratio of LC3-II/LC3-I was increased(P<0.05)in the SAH group.Compared with the SAH group,the expression level of Beclin-1 protein and the ratio of LC3-II/LC3-I was increased(P<0.05)in the drug group.Part II of the experiment:Western Blot results:Compared with the 3-MA(3-methyl adenine)group,the expression level of Beclin-1 protein and the ratio of LC3-II/LC3-I was increased in the3-MA+Xingnaojing injection group,the difference was statistically significant(P<0.05).Compared with the Rapa(rapamycin)group,the expression level of Beclin-1 protein and the ratio of LC3-II/LC3-I was increased in the Rapa+Xingnaojing injection group,the difference was statistically significant(P<0.05).Conclusions 1 Xingnaojing injection can moderately enhance autophagy of nerve cells in hippocampus of rats with subarachnoid hemorrhage and protect neurological function in rats with subarachnoid hemorrhage.2 Xingnaojing injection can antagonize the inhibitory effect of 3-MA(3-methyl adenine)on autophagy of nerve cells in hippocampus of rats with subarachnoid hemorrhage;Xingnaojing injection can synergize with Rapa(rapamycin)to activate autophagy of nerve cells in hippocampus of rats with subarachnoid hemorrhage.Figure 23;Table 5;Reference 105...