Protective Effect Of Jiawei Qifuyin And Zunyimycins On APP/PS1 Transgenic AD Mice

Objective:To study the effects of Jiawei Qifuyin and zunyimycins on learning and memory,neuroinflammation and intestinal flora in AD mice.The mechanism of Jiawei Qifuyin and zunyimycins exerting a protective effect on Alzheimer’s di sease was explored,and this paper provided a theoretical basis and experimental b asis for the subsequent guidance of the application of Jiawei Qifuyin and zunyimy cins to intervene in AD.Methods:（1）Network pharmacology was used to predict the active ingredient s,targets and potential signaling pathways of anti-AD of Jiawei Qifuyin.（2）The antioxidant activity of Jiawei Qifuyin in vitro was detected by enzy me tagging.（3）The CCK-8 method was used to determine the effect of Jiawei Qifuyin on the proliferative activity of Jurkat cells,Raji cells and macrophages and other immune cells in vitro.Lipopolysaccharide（LPS）was used to induce microglia（B V2）to establish a cell inflammation model,and ELISA experiments were used to detect the effect of Jiawei Qifuyin on the release of neuroinflammatory factors.（4）Morris water maze experiments were used to detect the effects of differ ent doses of Jiawei Qifuyin and zunyimycins on learning and memory ability of AD mice.（5）The effects of different drug interventions on the expression levels of ne uroinflammatory factors and pathway-related genes m RNA in AD mice were detec ted by q-PCR experiments.（6）16S r DNA was used to detect the changes of intestinal flora in AD mi ce after different drug interventions,and gas chromatography-mass spectrometry（G C-MS）was used to detect the changes of short-chain fatty acid content in the int estines of mice after different drug interventions.Results:（1）Network pharmacology results showed that the core targets of Ji awei Qifuyin were mainly AKT1 and TNF,and the AGEs/RAGE signaling pathway was one of the key pathways for its intervention in AD.（2）The results of in vitro activity experiments showed that Jiawei Qifuyin had certain antioxidant activity,and its hydroxyl radical scavenging rate and total antioxidant capacity had concentration-dependent effects;The Jiawei Qifuyin has a proliferative effect on immune cells,and the optimal concentration for Jurkat cell s is 5μg/m L,the optimal concentration for Raji cells is 1.25μg/m L,and the optim al concentration for macrophages is 10μg/m L.Different concentrations of Jiawei Q ifuyin can significantly reduce the release of neuroinflammatory factors IL-2,IL-6and TNF-αby LPS-induced BV2 cells.（3）The results of Morris water maze experiment showed that from the 2nd day of the positioning navigation experiment,the evasion latency period of mice in each group after drug intervention was statistically different from that of the blank control group（BC）（P<0.01）.（4）The Aβresults of mouse brain tissues in each group showed that comp ared with the BC group,the content of Aβ_1-40in the brain tissues of mice in eac h group after drug intervention was reduced（P<0.0001）;the content of Aβ_1-42in th e low,medium and high doses of Jiawei Qifuyin,sodium butyrate and nasal zunyi mycins intervention group was significantly reduced（P<0.01）.（5）The q-PCR results showed that the expression of IL-1βcould be downr egulated by low,medium and high doses of Jiawei Qifuyin compared with the B C group.Low-dose Jiawei Qifuyin,butyrate and zunyimycins-related intervention g roups could downregulate the expression level of IL-6;The expression of RAGE was downregulated in the low-dose Jiawei Qifuyin,butyrate,oral zunyimycins,an d nasal zunyimycins intervention groups.The expression levels of NF-κB were do wnregulated in the combination intervention group of donepezil,low-dose Qifuyin,and nasal zunyimycins.（6）The results of 16S r DNA showed that after different drug interventions,the intestinal microbiota abundance and intestinal microbiota function of each grou p of mice changed;The GC-MS results showed that the content of SCFAs in the intestines of each group of transgenic AD mice changed after different drug interv entions.Conclusions:（1）Jiawei Qifuyin has in vitro antioxidant,immune-enhancing a nd anti-neuroinflammatory activities.（2）Drugs such as Jiawei Qifuyin and zunyimycins can improve cognitive d ysfunction in AD mice and reduce Aβcontent in brain tissue of AD mice.（3）Drugs such as Jiawei Qifuyin and zunyimycins may reduce the neuroinf lammatory response in the brain of AD mice by regulating AGEs/RAGE/NF-κB a nd the gut-brain axis,thereby exerting a protective effect on AD.