| Purpose:Research learning and memory ability reduce MCI in heart kidney is not handed over to the performance level and mechanism,and based on the role of synaptic plasticity in learning and memory,On behalf of the formula Sagacious Confucius'Pillow Elixir(SCPE)is discussed in vivo and in vitro can improve glutamate function and mechanism of synaptic plasticity damage,in the hope of find MCI potential therapeutic targets,provide a reference basis for clinic.Methods:Part I:1.MTT method was used to detect the change of relative survival rate of cells after 24h and 48h of SH-SY5Y induced by A?25-35at 0,10,20,30,40,50?M concentration,and select the optimal concentration and time to establish cells in vitro model;2.CCK8 method was used to compare the effects of 10%,20%,30%SCPE-containing serum and 10%,20%,30%blank serum on the survival rate of A?25-35-induced SH-SY5Y cells at 24h and 48h,under light microscope Observe the effect of different concentrations of SCPE drug-containing serum on the morphology and axon length of SH-SY5Y cells induced by A?25-35,establish the optimal concentration and action time of SCPE drug-containing serum,and take the same concentration of blank serum as the subsequent experimental control group;3.According to the above experimental conditions,cells were divided into blank group?A?25-35model group?blank serum control+A?25-35group?blank serum+H-89+A?25-35group?blank serum+Sp-c AMP+A?25-35group?SCPE Serum+A?25-35group?SCPE serum+H-89+A?25-35group?SCPE serum+Sp-c AMP+A?25-35group.After 48 hours of incubation,the glutamate content of the cells upernatant was measured by the Elisa method;the protein expressions of PKA c-??p-CREB?BDNF?Glu A2?Glu N1and Glu N2A were detected by Western blot;the Synaptophysin and PSD-95 were detected by immunofluorescence protein.Part?:7-month-old SAMP8 mice were used as the research object,and SAMR1 mice of the same month-old were used as the blank control group.After 7 days of adaptive feeding,all mice were trained in Morris water maze,and 40 SAMP8 mice with cognitive impairment were selected and randomly divided into model group,SCPE high,medium and low dose groups,10 in each group;The other 10 SAMR1 mice were the blank control group.SCPE high,medium and low dose groups were given intragastric administration of SCPE suspension at the doses of 9.2g/kg·d?4.6g/kg·d?2.3g/kg·d,respectively,and mice in the model and blank groups were given the same volume Distilled water,the volume of administration is calculated by multiplying the body weight of the mouse by 0.1ml/10g,and it is administered intragastrically for 28 days,once a day.Before administration(day 0)and after administration(day 28),Morris water maze test was performed on mice in each group.Golgi-cox staining was used to observe the basic pathological changes in the hippocampal CA1 and CA3 areas of each group of mice;toluidine blue staining was used to observe the Nissl body status in the neurons of the hippocampal CA1 and CA3 areas of each group of mice;Western blot was used to detect the c AMP in the hippocampus of each group of mice,PKA c-??CREB?p-CREB?Glu A2?Glu N1 and Glu N2A protein expression.Result:Part I:1.Establish the optimal modeling concentration of A?25-35is30?M,the optimal concentration of SCPE drug-containing serum is 20?M,and the optimal action time is 48h;2.SCPE drug-containing serum may be improved by increasing expression of PKA-CREB-BDNF signaling pathway,thereby increasing the expression levels of glutamate synaptic receptor proteins Glu A2?Glu N1?Glu N2A,and synaptic plasticity-related proteins Synaptophysin?PSD-95,increasing intercellular synaptic glutamate release,improving cell morphology and Axon length.Part II:1.Zi Shen Ning Xin Hua Tan method can significantly improve the learning and memory ability of SAMP8 mice;2.Zi Shen Ning Xin Hua Tan method can significantly inhibit the loss of pyramidal neurons in the hippocampal CA1 region,increase the density of dendritic spines,improve the synaptic structural plasticity,and play a protective role in cognitive functions;3.Zi Shen Ning Xin Hua Tan method can improve the post-synaptic membrane thickness in glutamate synaptic structure,has an up-regulation effect on the expression of synaptic plasticity related proteins SYP and PSD-95,and has a protective effect on the normal performance of synaptic function;4.Zi Shen Ning Xin Hua Tan method showed up-regulation effect on the expression of c AMP-PKA,p-CREB/CREB and glutamate synaptic receptor proteins Glu A2?Glu N1 and Glu N2A in SAMP8 mice,suggesting that SCPE could promote the functional recovery and improve the transmission efficiency of MCI glutamate synaptic receptors.This effect may be regulated by the c AMP-PKA-CREB pathway.The above effects are dose-dependent,and the highest dose is the best in this study.In conclusion:... |
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