Effects Of Gubenjian Brain Method On NLRP3 Inflammasome And Related Inflammatory Factors In AD Model Rats

Objective To investigate the improvement effect and possible mechanism of Gubenjian brain method on NLRP3 inflammasome and related inflammatory factors in AD model rats.Methods: Sixty SD male rats were subjected to adaptive feeding for one week,and they were divided into 6 groups each of blank group,model group,low-dose group,medium-dose group,high-dose group and positive drug group(donepezil)by random group,with 10 rats in each group.The blank group was not treated,and the model group,the low-dose group of Gubenjian cerebral fluid,the medium-dose group of Gubenjian cerebral fluid,the high-dose group of Gubenjian cerebral fluid,and the positive drug group(donepezil)were injected with Aβ1-42 in the CA1 region of the hippocampus to establish an AD rat model.Starting from the 4th day after the end of modeling,the low-dose group of Gubenjian cerebral fluid,the medium-dose group of Gubenjian cerebral fluid,and the high-dose group of Gubenjian cerebral fluid were given Gubenjian brain formula for gastric gavage,and the positive drug group(donepezil)was given donepezil powder for gastric gavage,and after 4 weeks of continuous gavage,the learning and memory ability of rats was detected by water maze experiment.24 h after the last administration,tissues were taken and HE staining was used to detect the morphological function of neurons in the CA1 region of the hippocampus in each group.Nicholferi staining analyzed the change of Nicholian body number in the CA1 region of the hippocampus in each group.Immunohistochemical detection of Aβ1-42 in the CA1 region of the rat hippocampus;The Western blot method detected the expression levels of NLRP3,ASC,Caspase-1 and IL-18 proteins in rat hippocampal tissues.ELISA detected the levels of inflammatory factors IL-1 β,TNF-α and IL-6 in rat brain tissue.Results 1.The water maze experiment confirmed that in the directional navigation test,the rats in the normal group had significantly shorter escape latency than the model group(P<0.05),and the rats in the model group had significantly longer evasion latency than the low-dose group,the medium-dose group,the high-dose group,and the positive drug group(donepezil)(P<0.05),and the movement trajectory was disorderly.In the space exploration experiment,compared with the normal group,the number of rats crossing the target quadrant platform and the target quadrant residence time in the model group were significantly reduced(P<0.05),while the number of rats crossing the platform(except the low-dose group of solid benjian cerebral fluid)and the target quadrant residence time of rats in each drug group was significantly increased(P<0.05)compared with the model group.2.Nicholich staining experiments confirmed that the nuclei of the normal group were complete,neatly arranged,and the number of neurons was abundant.The nuclei of the model group were incomplete,arranged and scattered,and the number of nylonite in the cell was scarce.Compared with the model group,the number of neuronal cells in rats in each administration group was less damaged,the morphology of nuclei was significantly improved,and the arrangement of nyinite was relatively orderly,uniform and tight.3.HE staining showed that the normal group had the largest number of hippocampal neurons and complete morphology,while the number of model groups was significantly reduced,the arrangement was sparse and scattered,and the number of neuronal cells in each treatment group increased compared with the model group,and the cell arrangement was relatively neat and uniform.4.Western blot showed that the expression of NLRP3,ASC,Caspase-1 and IL-18 proteins in the model group was higher than that in the blank group(P<0.05),and compared with the model group,the expression of NLRP3,ASC,Caspase-1 and IL-18 proteins in the remaining groups decreased(P<0.05).5.ELISA showed that the levels of inflammatory factors IL-1β,TNF-α and IL-6 in the model group were significantly higher than those in the blank group(P<0.05),and the levels of inflammatory factors IL-1β,TNF-α and IL-6 in the high,medium and low groups of Gubenjian cerebral fluid were significantly reduced(P<0.05)compared with the model group.Conclusion The mechanism of Aβ1-42-induced learning and memory in AD rats may be related to NLRP3 inflammasome and related inflammatory factors.