Improvement Of Cognitive Function In Alzheimer Mice By Jiedu Yizhi Formula Inhibition Of Neuroinflammation Through IL-17-p53 Pathway

Objective:To investigate the effects of Jiedu Yizhi formula（JDYZ）and its active ingredientβ-sitosterol（BS）on cognitive function in Alzheimer’s model mice and to elucidate the possible molecular mechanisms.Methods:Prediction of potential active ingredients and signal pathways for the treatment of AD with Jiedu Yizhi formula by networkpharmacology analysis.Behavioral observation experiment 1:C57BL/6J mice were randomly divided into control,SCOP,BS-L and BS-H groups,8 mice in each group.The acute dementia model in mice was established by intraperitoneal injection of scopolamine with pretreatment byβ-sitosterol.The improvement effect ofβ-sitosterol on short-term memory in demented mice was assessed by Y-maze test.Behavioral observation experiment 2:C57BL/6J mice were randomly divided into control,sham,AD,DNPZ,JDYZ-L,JDYZ-H,BS-L and BS-H groups,10 mice in each group.We established an AD model by injecting oligomeric amyloidβ(Aβ_1-42)into the lateral ventricle of mice,intervened withβ-sitosterol for treatment.We systematically assessed the protective effects ofβ-sitosterol on AD mice through behavioral observation experiments such as nesting behavior,new object discrimination and Morris water maze test.The expression levels of IL-17,p53 and apoptosis-related proteins Bax,Bcl-2,Caspase-3 and Cleaved Caspase-3 in the hippocampal tissues of mice were detected by Realtime q PCR（RT-q PCR）and Western blotting.The levels of inflammatory cytokines and oxidative stress-related SOD and GSH activities in hippocampal tissues were examined.Results:The potential active component of the Jiedu Yizhi formula for AD isβ-sitosterol,which is closely related to the IL-17-p53 signaling pathway.In behavioral observation experiment 1,the rate of spontaneous alternation was significantly higher in mice pretreated withβ-sitosterol（P<0.05）,and the rate of exploration of the novel arm was elevated（P>0.05）.In behavioral observation experiment 2,mice treated with JDYZ or BS exhibited significantly higher nesting behavior scores（P<0.05）,prolonged novel object recognition（P<0.01 or P<0.05）,fewer instances of escape latency during platform trials and a smaller number of platform crossings（P<0.01 or P<0.001）versus the AD group in behavioral observation experiments.RT-q PCR detected the IL-17A and p53 m RNA expression,which were significantly upregulated in the AD group（P<0.01）and JDYZ or BS treated group were downregulated（P<0.05 or P<0.01）.Simultaneously,compared with the AD group,the expression of IL-17A,p53,and apoptosis-related proteins Bax and Cleaved Caspase-3 was significantly decreased（P<0.01）and Bcl-2 expression was increased（P<0.01）in the JDYZ or BS treated group.TNF-αand IL-17A levels were remarkable decreased（P<0.05 or P<0.01）,SOD activity and GSH content were remarkable increased（P<0.01 or P<0.001）in the JDYZ or BS treated group.Conclusion:This study elucidates that Jiedu Yizhi formula and its active ingredientβ-sitosterol may inhibit the IL-17-p53 signaling pathway to reduce neuroinflammation and inhibit apoptosis and oxidative stress,thus improving cognitive function in AD mice.