The Research Of Effect And Mechanism Of HuangjingWan On Treating The Mice With Alzheimer’s Disease

Objective:Based on the previous studies,the repeatability of Huangjing Wan in the treatment of Alzheimer’s disease（AD）,and which the possible mechanism in the treatment of AD was explored by this experiment.Methods:1.Animals and groups:140 male Kunming mice with SPF grade,body weight（26±2）g,were randomly divided into normal control group,sham operation control group,AD model group,positive drug group,low,medium and high-dose group of Huangjing Wan(equivalent to 1.25,2.5,7.5 g·kg^-1·d^-1),with 10 mice in each group.All the animals were divided into Experiment 1 and Experiment 2.2.AD modeling and experimental drug intervention:No1 experiment:The mice of AD model group and all experimental drug groups were given intraperitoneal injection of 1%D-galactose(0.14 g.kg^-1.d^-1)solution which prepared with sterile0.9%Na Cl solution,mice of sham operation control group were given with the equal volume of sterile 0.9%Na Cl solution at the sanme site.4 weeks after continuous injection,mice of AD model group and experimental drug group were given one-time right ventricular injection with 1μg Aβ_1-42to establishe model control,and mice of the sham operation control group were injected with 1μL normal saline in right ventricular to make the model control.14 days after the establishment of AD model,mice of all experimental drug groups were received corresponding drugs and dose through intragastric administration,while mice of sham operation control group and AD model group were given the same dose of sterile 0.9%Na Cl solutionthrough intragastric administration,once a day for 28 days;Normal control group mice without special disposal,only daily feeding.No2 experiment:the mice of AD model group and experimental drug group were given intraperitoneal injection with 1%D-galactose(0.14 g.kg^-1.d^-1)solution which prepared with sterile 0.9%Na Cl solution,the mice were injected continuously for 21 days to induce aging;On the 22nd day,scopolamine(2.0×10^-3g·kg^-1·d^-1)was injected intraperitoneally for 14 days to induce AD dementia in mice.The mice in the sham operation control group were given the same volume of sterile 0.9%Na Cl solution once a day.The mice in the normal control group were not injected.3.Detective methods:learning and memory abilities of each group mice were evaluated by Platform Jumping Test.HE staining and Nissl staining were used to detect the number changes of the cerebral cortex and hippocampal neurons in each group mice.The activities changes of SOD and GSH-Px in brain tissue and serum of all mice were detected by colorimetry;ELISA was used to detect differences in the levels of inflammatory factors such as IL-1βand TNF-αin brain tissue and serum of all mice.ELISA and WB were used to detect the changes of Aβ_1-42protein expression in brain tissue of all mice;Brd U immunohistochemistry and Brd U immunofluorescence methods were used to detect the proliferation of neural stem cells（NSCs）in the dentate gyrus（DG）area of the hippocampus of all mice.Results:1.Detection results of behavioral examination:Compared with the normal group,the learning and memory ability of AD model group was significantly decreased,the dementia state was more obvious in AD group（Respectively P<0.01）.Compared with the AD model group,The dementia symptoms of mice in each dose group of Huangjing Wan were significantly improved,significantly improve the achievement of learning and memory（P<0.05,P<0.01）;In the dose range of 1.25-7.5 g.kg^-1.d^-1,and with the increase of the dose of Huangjing Wan,the more obvious its effect.2.Detection results of biochemical indexes:Compared with the normal group,The activities of SOD and GSH-Px in brain tissue and serum were significantly reduced,The levels of IL-1βand TNF-αinflammatory factors increased significantly in AD group（Respectively P<0.01）.Compared with the AD model group,the activities of SOD and GSH-Px in brain tissue and serum of mice in each dose group of Huangjing Wan were increased,and the levels of inflammatory factors IL-1βand TNF-αwere decreased（P<0.05,P<0.01）,In the dose range of 1.25-7.5 g.kg^-1.d^-1,and with the increase of the dose of Huangjing Wan,the more obvious its effect.3.Detection results of Aβ_1-42protein content in brain tissue:Compared with the normal group,the content of Aβ_1-42protein in brain tissue is significantly increased in AD group（Respectively P<0.01）.Compared with the AD model group,the content of Aβ_1-42protein in brain tissue of mice in each dose group of Huangjing Wan was significantly decreased（P<0.05,P<0.01）,In the dose range of 1.25-7.5 g.kg^-1.d^-1,and with the increase of the dose of Huangjing Wan,the more obvious its effect.4.Detection results of neuron cell numbers in the cerebral cortex and hippocampal CA1 and CA3 regions:Compared with the normal group,the neuron cell numbers in the cerebral cortex and hippocampal CA1 and CA3 regions were decreased in AD group（Respectively P<0.01）.Compared with the AD group,the neuron cell numbers in the cerebral cortex and hippocampal CA1 and CA3 regions of mice in each dose group of Huangjing Wan were increased（P<0.05,P<0.01）;In the dose range of 1.25-7.5 g.kg^-1.d^-1,and with the increase of the dose of Huangjing Wan,the more obvious its effect.5.Brd U positive cell detection results of NSCs in hippocampal DG region:Compared with the normal group,the Brd U positive cell numbers of NSCs in hippocampus DG area were decreased（Respectively P<0.01）.Compared with the AD model group,the Brd U positive cell numbers of NSCs in hippocampus DG area of mice in each dose group of Huangjing Wan were increased（P<0.05,P<0.01）;In the dose range of 1.25-7.5 g.kg^-1.d^-1,and with the increase of the dose of Huangjing Wan,the more obvious its effect.Conclusions:1.Huangjing Wan has a good effect on the treatment of AD,and its pharmacodynamic function is repeatable and stable.2.The roles of Huangjing Wan in preventing and treating AD are related to its anti-oxidation and anti-inflammatory,alleviating central nervous oxidative stress,reducing Aβprotein deposition,maintaining the stability of the central nervous environment and promoting the proliferation of hippocampal NSCs.