Effect of Early Intervention of Yishen Huazhuo Prescription on Learning and Memory of Accelerated Aging SAMP8 Mice and Its Mechanism / 中国实验方剂学杂志

ObjectiveTo investigate the impact of early intervention with Yishen Huazhuo prescription （YHP） on the learning and memory of accelerated aging model mice， as well as its underlying mechanism. MethodForty-eight 3-month-old male SAMP8 mice were randomly assigned into four groups， including the model group， low-dose YHP group， high-dose YHP group， and donepezil group. Additionally， 24 SAMR1 mice of the same age were divided into a control group and a YHP treatment control group， each consisting of 12 mice. The YHP groups received YHP at doses of 6.24 g·kg-1 and 12.48 g·kg-1， while the donepezil group was treated with donepezil at a dose of 0.65 mg·kg-1. The model group and control groups were given physiological saline. The mice were gavaged once daily for a duration of four weeks. Spatial learning and memory abilities of mice were assessed using the Morris water maze test. Immunofluorescence staining was employed to evaluate neuronal density as well as expression levels of M1 microglial （MG） polarization marker inducible nitric oxide synthase （iNOS） and M2 MG polarization marker arginase-1 （Arg-1） in the hippocampus region. Enzyme-linked immunosorbent assay （ELISA） was used to measure serum levels of pro-inflammatory factor interleukin 1β （IL-1β） and anti-inflammatory factor transforming growth factor-β1 （TGF-β1）. Furthermore， Western blot analysis was conducted to determine expressions of amyloid β peptide1-42 （Aβ1-42） along with triggering receptor expressed on myeloid cells 2 （TREM2）/nuclear factor kappa B （NF-κB） signaling pathway-related proteins TREM2， phospho （p）-NF-κB p65， and phospho-inhibitory kappa B kinase β （IKKβ） in the hippocampus. ResultCompared with the control group， the model group exhibited a significantly prolonged escape latency （P<0.01）， a significant reduction in neuron-specific nuclear protein （NeuN） expression in the hippocampus， a significant increase in iNOS expression in MG， and a significant decrease in Arg-1 expression. The serum IL-1β content was significantly increased， while the TGF-β1 content was significantly decreased. Additionally， there was a significant decrease in TREM2 expression in the hippocampus and significant increases in p-NF-κB p65， p-IKKβ， and Aβ1-42 expressions （P<0.05， P<0.01）. However， no significant changes were observed in escape latency， times of crossing the platform， and hippocampal NeuN expression in the YHP treatment control group. Conversely， iNOS expression in MG as well as the hippocampal p-NF-κB p65， p-IKKβ， and Aβ1-42 expressions were significantly decreased. Furthermore， TREM2 expression was significantly increased （P<0.05， P<0.01）. In comparison to the model group， the low-dose YHP group showed a significantly shortened escape latency and an increased number of crossing the platform （P<0.05， P<0.01）. In the high-dose YHP group， the escape latency was significantly shortened （P<0.05）. In the low-dose YHP group， high-dose YHP group， the expression of NeuN in the hippocampus was significantly increased， the expression of iNOS in MG was significantly decreased， and the expression of Arg-l was significantly increased. The serum IL-1β content was significantly decreased， while the TGF-β1 content was significantly increased. Furthermore， the expression of TREM2 in the hippocampus was significantly increased， and the expressions of p-NF-κB p65， p-IKKβ， and Aβ1-42 were significantly decreased （P<0.01）. ConclusionEarly YHP intervention may promote the transformation of hippocampal MG from M1 to M2 by regulating the TREM2/NF-κB signaling pathway， reduce the release of neuroinflammatory factors， protect hippocampal neurons， and reduce the deposition of Aβ1-42， and finally delay the occurrence of learning and memory decline in SAMP8 mice.

Xixin Decoction improves learning and memory ability of SAMP8 by enhancing neuroprotective effect and inhibiting neuroinflammation / 中国中药杂志

This study aimed to explore the possible effect of Xixin Decoction(XXD) on the learning and memory ability of Alzheimer's disease(AD) model senescence-accelerated mouse-prone 8(SAMP8) and the related mechanism in enhancing neuroprotective effect and reducing neuroinflammation. Forty SAMP8 were randomly divided into a model group(10 mL·kg~(-1)·d~(-1)), a probiotics group(0.39 g·kg~(-1)·d~(-1)), a high-dose group of XXD granules(H-XXD, 5.07 g·kg~(-1)·d~(-1)), a medium-dose group of XXD granules(M-XXD, 2.535 g·kg~(-1)·d~(-1)), and a low-dose group of XXD granules(L-XXD, 1.267 5 g·kg~(-1)·d~(-1)). Eight senescence-accelerated mouse-resistant 1(SAMR1) of the same age and strain were assigned to the control group(10 mL·kg~(-1)·d~(-1)). After ten weeks of intragastric administration, the Morris water maze was used to test the changes in spatial learning and memory ability of mice after treatment. Meanwhile, immunofluorescence staining was used to detect the positive expression of receptor for advanced glycation end products(AGER), Toll-like receptor 1(TLR1), and Toll-like receptor 2(TLR2) in the hippocampal CA1 region of mice. Western blot was employed to test the protein expression levels of silencing information regulator 2 related enzyme 1(SIRT1), AGER, TLR1, and TLR2 in the hippocampus of mice. Enzyme linked immunosorbent assay(ELISA) was applied to assess the levels of Aβ_(1-42) in the hippocampus of mice and the levels of nuclear factor κB p65(NF-κB p65), NOD-like receptor protein 3(NLRP3), tumor necrosis factor-α(TNF-α), and interleukin-1β(IL-1β) in the serum and hippocampus of mice. Compared with the model group, XXD significantly improved the spatial learning and memory ability of SAMP8, increased the expression of neuroprotective factors in the hippocampus, decreased the levels of neuroinflammatory factors, and inhibited the expression of Aβ_(1-42). In particular, H-XXD significantly increased the expression of SIRT1 in the hippocampus of mice, reduced the expression levels of NF-κB p65, NLRP3, TNF-α, and IL-1β in the serum and hippocampus of mice, and decreased the expression of AGER, TLR1, and TLR2 in the hippocampus of mice(P<0.05 or P<0.01). XXD may improve the spatial learning and memory ability of AD model SAMP8 by enhancing the neuroprotective effect and inhibiting neuroinflammation.

Effect of human urinary kallidinogenase on cognitive function of SAMP8 mouse model and its mechanism / 中国药理学通报

Aim To study the effect of human urinary kallidinogenase(HUK)on the cognitive function of SAMP8 mouse model and its mechanism. Methods SAMP8 mice were divided intofive groups:SAMP8 group,treatment group(giving 8.75×10-3,1.75×10-2,3.5×10-2,7.0×10-2 HUK),and the SAMR1 vehicle group was used as blank control. Each group was performed Morris water maze to detect spatial cognition. Afterwards the group with the most obvious cognitive improvement(HUK group)was selected for the follow-up experiments. Immunohistochemical detection of ChAT expression in CA3 area was further verified by RtPCR. Western blot was used to detect the expression of PSD95,SYN,BDNF,and pCREB protein. The activity of MPO and the content of IL-1β and IL-18 were determined. Results The passing times in the SAMP8 group was less than that of the SAMR1 group(P<0.05). The passing times of treatment group increased compared with the SAMP8 group(P<0.05 or P<0.01),and the spatial probe time of the target quadrant was shorter(P<0.05 or P<0.01). We conducted follow-up experiments with group d(HUK group). The expression of ChAT positive cells in CA3 area of SAMP8 group was significantly lower than that of SAMR1 group; the expression of positive cells in HUK group significantly increased; RtPCR showed that ChAT expression in SAMP8 group was lower than that in SAMR1 group,and ChAT expression was significantly higher than that in SAMP8 group after HUK treatment. Compared with the SAMR1 group,the levels of IL-1β,IL-18 and MPO activity in the CA3 area of SAMP8 group significantly increased,and the protein expressions of PSD95,SYN,BNDF and pCREB decreased. After HUK treatment,the content of IL-1β,IL-18 and MPO activity decreased,and the expression of PSD95,SYN,BNDF and pCREB increased. Conclusions HUK can improve the spatial cognition of SAMP8 mice. The mechanism may be achieved by promoting the expression of ChAT in CA3 area,reducing the oxidative stress and increasing synapse-related proteins.

Effects and mechanisms of melatonin combined with enriched environment on learning and memory ability of SAMP8 mice / 中华行为医学与脑科学杂志

Objective:To explore the effects and possible mechanisms of melatonin combined with enriched environment on the learning and memory ability of senescence-accelerated mouse prone 8(SAMP8).Methods:Forty-eight SAMP8 male mice aged 4 months were randomly divided into model group, enriched environment group, melatonin group and melatonin combined with enriched environment group (combined intervention group) by random number table method, with 12 mice in each group. Mice in the melatonin group and combined intervention group were subcutaneously injected with melatonin at a dose of 8 mg·kg -1·d -1, and the mice in the model group and the enriched environment group were given the same amount of normal saline instead.The mice in model group and melatonin group were raised in a standard environment, and the mice in enriched environment group and combined intervention group were raised in an enriched environment.The intervention lasted 28 days. The aging degree of mice was scored before and 28 days after the intervention. Morris water maze test was used to detect the learning and memory ability of mice. Nissl staining and TUNEL staining were used to observe the Nissl staining positive cells and apoptotic cells in the CA1 area of hippocampus.ELISA was used to detect the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the hippocampus of mice. Western blot was used to detect the levels of amyloid β-protein (Aβ) 1-42, microtubule-associated protein tau (tau) phosphorylated at threonine (Thr) 205 (Tau pT205), Toll-like receptor 4 (TLR4), and nuclear factor-κB (NF-κB) p65 protein in the hippocampus of mice. qRT-PCR was used to detect the levels of TLR4, NF-κB p65 mRNA in the hippocampus of mice. SPSS 22. 0 statistical software was used for repeated measure ANOVA, one-way ANOVA and LSD test. Results:(1) Aging score: after intervention, the aging scores of mice in the four groups were significantly different ( F=120.601, P<0.01). The aging scores of mice in the enriched environment group, melatonin group, and combined intervention group were lower than those in the model group (all P<0.05), while the aging score of mice in the combined intervention group was significantly lower than those in the enriched environment group and melatonin group (both P<0.05). (2) The results of the location navigation experiment showed that the time × group interaction effect of the escape latencies of mice in the four groups were significant ( F=30.524, P<0.001). From the 2nd to 4th day, the escape latencies of mice in the enriched environment group, melatonin group and combined intervention group were all lower than that in the model group (all P<0.05). The results of the space exploration experiment showed that the residence time in the target quadrant and the number of platform crossings of mice in the four groups were significantly different ( F=291.328, 113.482, both P<0.01). The residence time in the target quadrant ((29.45±1.70)s, (32.44±1.55)s, (37.48±0.84) s) and the number of platform crossings ((6.44±0.61) times, (7.16±0.70) times, (12.60±1.23) times) of mice in the enriched environment group, melatonin group and combined intervention group were higher than those in the model group ((15.07±1.28) s, (4.10±0.61) times), while the residence time in the target quadrant and the number of platform crossings of mice in the enriched environment group and the melatonin group were significantly lower than those in the combined intervention group (all P<0.05). (3) Nissl and TUNEL staining showed that the number of Nissl positive neurons in the hippocampal CA1 region of mice in the four groups were significantly different ( F=809.264, P<0.01), and the number of apoptotic cells in the hippocampal CA1 region were also significantly different ( F=1 060.583, P<0.01). The number of Nissl stained positive neurons in the hippocampal CA1 region of mice in the combined intervention group was more than those in the model group, enriched environment group, and melatonin group (all P<0.05), and the number of apoptotic cells were less than those in the model group, enriched environment group, and melatonin group (all P<0.05). (4) The results of ELISA assay showed that there were significantly different in the levels of IL-1β, IL-6 and TNF-α in the hippocampus of mice in the four groups ( F=152.887, 63.506, 432.026, all P<0.01). The contents of IL-1β, IL-6 and TNF-α in the hippocampus of mice in the enriched environment group, melatonin group, and combined intervention group were lower than those in the model group(all P<0.05). Among them, the contents of IL-1β, IL-6 and TNF-α in the hippocampus of mice in the enriched environment group and melatonin group were significantly higher than those in the combined intervention group (all P<0.05). (5) Western blot analysis showed that there were significantly different in the protein expression levels of Aβ1~42, tau pT205, TLR4, NF-κB p65 in the hippocampus of mice in the four groups ( F=122.349, 98.934, 201.635, 116.553, all P<0.01). The protein expression levels of Aβ1-42, tau pT205, TLR4, and NF-κB p65 in the hippocampus of mice in the enriched environment group, melatonin group, and combined intervention group were lower than those in the model group.Among them, the protein expression levels of Aβ1-42, tau pT205, TLR4, NF-κB p65 in the hippocampus of mice in the enriched environment group and melatonin group were significantly higher than those in the combined intervention group (all P<0.05). (6) qRT-PCR showed that the mRNA expression levels of TLR4 and NF-κB p65 in the hippocampus of mice in the four groups were significantly different ( F=42.913, 102.446, both P<0.01). The mRNA expression levels of TLR4 ((0.63±0.05), (0.55±0.04), (0.42±0.03)) and NF-κB p65 ((0.98±0.06), (0.82±0.04), (0.72±0.04)) in the hippocampus of mice in the enriched environment group, melatonin group and combined intervention group were lower than those in the model group ((0.74±0.07), (1.20±0.05)) (all P<0.05). Among them, the mRNA expression levels of TLR4 and NF-κB p65 in the hippocampus of mice in the enriched environment group and melatonin group were significantly higher than those in the combined intervention group (all P<0.05). Conclusion:Melatonin combined with enriched environment can improve the learning and memory ability and neuroinflammatory response of SAMP8 mice, and its mechanism may be related with the down-regulation of TLR4/NF-κB p65 signaling pathway.

Haikun Shenxi capsule improved cognitive dysfunction by activating autophagy in SAMP8 mice / 中国药理学通报

Aim To study the modulation effects of HKSX capsule on Alzheimer's disease (AD) and to preliminarily explore its mechanism using SAMP8 as an AD model.Methods SAMP8 mice aged 4 months were randomly divided into model group ( P8 group) , HKSX low-dose group (L-HKSX group) , HKSX high- dose group ( H-HKSX group) , and senescence-accel- erated mouse/resistance 1 (SAMR1 ) at the same age was used as normal control group ( R1 group).New object recognition and Morris water maze experiments were used to detect the learning and memory abilities of each group.Levels of A(3, _40 and A(3, 42 were detected by ELISA, and the expression levels of LC3- U , p62, Beclin-1 , PSD95 and Syn in hippocampus of each group were detected by Western blot.Results Compared with model group, both low and high doses of HKSX could enhance the D1 in new object recognition test, increase number of crossing the platform anrl the time spent in the target quadrant, and shorten the escape latency.Besides, it also enhanced the clearance °f Ap, _40 and AfJ, _42 , up-regulated the relative expression of Beclin-1 and LC3- II and down-regulated the expression of P62.In addition, it increased the expression of synaptic-related proteins PSD95 and Syn.Conclusions HKSX capsule can regulate the expression of autophagy-related proteins and improve autoph- agv dysfunction, which in turn reduce the deposition of A(3 in vivo to alleviate its cytotoxicity and improve synaptic plasticity.Thus, HKSX can improve the learning and memory deficits of AD mice.

Effect of early intervention of electroacupuncture on learning-memory ability and level of hippocampal phosphorylated Tau protein in SAMP8 mice / 中国针灸

OBJECTIVE@#To explore the effect of early intervention electroacupuncture (EA) at "Baihui" (GV 20), "Dazhui" (GV 14) and "Shenshu" (BL 23) on the learning-memory ability and the expression of phosphorylated Tau protein in the hippocampus of SAMP8 mice, so as to provide reference for the intervening period of EA for Alzheimer's disease (AD).@*METHODS@#A total of 36 3-month old SAMP8 mice were randomly divided into a model group, a 3-month-old EA group and a 9-month-old EA group, 12 mice in each group. Twelve normal SAMR1 mice with the same age were taken as the control group. The mice in the 3-month-old EA group and 9-month-old EA group were treated with EA at "Baihui" (GV 20), "Dazhui" (GV 14) and "Shenshu" (BL 23) separately 3 months old and 9 months old (continuous wave, 2 Hz, 1.5-2 mA), 20 min each time, once a day, 8 days as a course of treatment, with an interval of 2 days between courses, totally 3 courses of treatment were given. The mice sample in each group was collected at the age of 10 months after the learning-memory ability tested by Morris water maze. The expression of phosphorylated Tau protein in the hippocampus was detected by immunohistochemistry and Western blot, and the expression of Tau mRNA was detected by real-time PCR.@*RESULTS@#Compared with the control group, in the model group, the escape latency was significantly increased (<0.01), the time of stay in the original platform quadrant and the number of crossing the platform quadrant were reduced (<0.01), and the expressions of phosphorylated Tau protein and Tau mRNA in hippocampus were increased (<0.01). Compared with the model group, in the 3-month-old EA group and 9-month-old EA group, the escape latency was significantly reduced (<0.05), the time of stay in the original platform quadrant and the number of crossing the platform quadrant were increased (<0.05), and the expressions of phosphorylated Tau protein and Tau mRNA in hippocampus were reduced (<0.05). Compared with the 9-month-old EA group, in the 3-month-old EA group, the escape latency was significantly reduced (<0.05), the time of stay in the original platform quadrant and the number of crossing the platform quadrant were increased (<0.05), and the expressions of phosphorylated Tau protein and Tau mRNA were reduced (<0.01).@*CONCLUSION@#The early EA intervention could more effectively improve the learning-memory ability and inhibit phosphorylation of Tau protein in the hippocampus of SAMP8 mice.

Transcriptome analysis of the hippocampus in environmental noise-exposed SAMP8 mice reveals regulatory pathways associated with Alzheimer's disease neuropathology

BACKGROUND@#Chronic noise exposure is one environmental hazard that is associated with genetic susceptibility factors that increase Alzheimer's disease (AD) pathogenesis. However, the comprehensive understanding of the link between chronic noise stress and AD is limited. Herein, we investigated the effects of chronic noise exposure on AD-like changes in senescence-accelerated mouse prone 8 (SAMP8).@*METHODS@#A total of 30 male SAMP8 mice were randomly divided into the noise-exposed group, the control group, and aging group (positive controls), and mice in the exposure group were exposed to 98 dB SPL white noise for 30 consecutive days. Transcriptome analysis and AD-like neuropathology of hippocampus were examined by RNA sequencing and immunoblotting. Enzyme-linked immunosorbent assay and real-time PCR were used to further determine the differential gene expression and explore the underlying mechanisms of chronic noise exposure in relation to AD at the genome level.@*RESULTS@#Chronic noise exposure led to amyloid beta accumulation and increased the hyperphosphorylation of tau at the Ser202 and Ser404 sites in young SAMP8 mice; similar observations were noted in aging SAMP8 mice. We identified 21 protein-coding transcripts that were differentially expressed: 6 were downregulated and 15 were upregulated after chronic noise exposure; 8 genes were related to AD. qPCR results indicated that the expression of Arc, Egr1, Egr2, Fos, Nauk1, and Per2 were significantly high in the noise exposure group. These outcomes mirrored the results of the RNA sequencing data.@*CONCLUSIONS@#These findings further revealed that chronic noise exposure exacerbated aging-like impairment in the hippocampus of the SAMP8 mice and that the protein-coding transcripts discovered in the study may be key candidate regulators involved in environment-gene interactions.

Metabolomics-based study of metabolic changes in liver tissues of senescence accelerated mouse prone 8 (SAMP8) treated with baicalein / 中草药

Objective: To study the protective effect and explore the mechanism of baicalein on the liver of senescence-accelerated mouse prone 8 (SAMP8) based on 1H-NMR metabolomics. Methods: The protective effect of baicalein (ig) on the liver of SAMP8 mice was investigated in the present study. The mice in control group was SAMR1, the mice in model group was SAMP8, the drug treatment group was SAMP8 + baicalein. The mice in control and model group were administrated with equal amount of normal saline, and the mice in drug treatment group were administrated with 200 mg/kg baicalein. The liver tissues of mice in each group were isolated, and the damage degree of liver tissue was determined by hematoxylin-eosin (HE) staining. 1H-NMR combined with multivariate statistical analysis was used to investigate the mechanism of baicalein on liver damage in aging mice. Results: Organ index and HE staining results showed that baicalein can significantly improve liver damage in SAMP8 mice. Eight potential biomarkers were found in hepatic metabolomics analysis, mainly involving three metabolic pathways: Alanine, aspartic acid and glutamate metabolism; Glycine, serine, threonine metabolism, and inositol phosphate metabolism. Conclusion: The study of metabolites alterations in the liver tissue of SAMP8 mice would provide experimental evidence for anti-aging drug research.

Effects of acteoside on learning,memory and neurotransmitters level in brain tissue of SAMP8 mice / 中华行为医学与脑科学杂志

Objective To evaluate the effect of acteoside on learning,memory and neurotransmitter in SAMP8 mice. Methods The 6-month-old rapidly aging SAMP8 mice were randomly divided into model group,namenda group,low-dose acteoside group(30 mg·kg-1 ·d-1),medium-dose acteoside group(60 mg ·kg-1 ·d-1) and high-dose acteoside group(120 mg·kg-1 ·d-1 ) according to the digital table method, with 12 in each group. And 12 SAMR mice with the same age resistance were used as the control group. After 75 days of continuous intragastric administration,Morris water maze method and spontaneous activity experi-ment were used to investigate the effects of acteoside on learning,memory and anxiety of mice. The levels of neurotransmitters acetylcholine ( ACh), serotonin ( 5-HT ), norepinephrine ( NE ) and dopamine ( DA ) in mouse brain tissue(cortex and hippocampus) were detected by ELISA. Results (1) In the Morris water maze test,compared with the model group,the acteoside significantly reduced the escape latency of SAMP8 mice in training period. (2)In the experiment of autonomic activity,compared with the model group,the aver-age speed and total distance of the low-dose acteoside group were significantly increased(t=15. 0,20. 8,both P<0. 05);the number of vertical movements and the average speed of the medium-dose acteoside group were significantly increased(t=15. 8,13. 6,both P<0. 05). The average speed and total distance of the high-dose acteoside group were remarkarbly increased(t=30. 9,29. 7,both P<0. 05),and the number of defecations in the mice of the lav-dose acteoside group, medium-dose acteoside group, high-dose acteoside group were ((2. 83±1. 19)particles,(3. 25± 1. 29) particles,(2. 58± 1. 16) particles),they were obviously lower than that of the model group ((5. 25±1. 48) particles)(t=15. 7,20. 1,13. 5,all P<0. 01). (3) Simultaneously, the contents of ACh,NE and DA in the hippocampus and cortex of the model group were significantly lower than those in the normal group (hippocampus:t=10. 3,12. 7,13. 2,all P<0. 05;cortex:t=11. 7,10. 5,12. 4, all P<0. 05). Compared with the model group,the contents of ACh,NE,DA and 5-HT in the hippocampus and cortex of the low,medium and high doses of acteoside were significantly increased ( hippocampus: t=31. 4, 20. 3,10. 7,12. 9,all P<0. 05;cortex:t=33. 7,29. 4,14. 5,12. 7,all P<0. 05). Conclusion The ac-teoside can enhance the ability of spatial learning and memory of SAMP8 mice,and can regulate the depres-sion and anxiety of animals. The mechanism may be related to the ACh content and the monoamine neuro-transmitters increase in the brain.

Effects of acteoside on learning, memory and neurotransmitters level in brain tissue of SAMP8 mice / 中华行为医学与脑科学杂志

Objective@#To evaluate the effect of acteoside on learning, memory and neurotransmitter in SAMP8 mice.@*Methods@#The 6-month-old rapidly aging SAMP8 mice were randomly divided into model group, namenda group, low-dose acteoside group(30 mg·kg-1 ·d-1), medium-dose acteoside group(60 mg·kg-1 ·d-1) and high-dose acteoside group(120 mg·kg-1 ·d-1) according to the digital table method, with 12 in each group.And 12 SAMR mice with the same age resistance were used as the control group.After 75 days of continuous intragastric administration, Morris water maze method and spontaneous activity experiment were used to investigate the effects of acteoside on learning, memory and anxiety of mice.The levels of neurotransmitters acetylcholine(ACh), serotonin(5-HT), norepinephrine(NE) and dopamine(DA) in mouse brain tissue(cortex and hippocampus) were detected by ELISA.@*Results@#(1)In the Morris water maze test, compared with the model group, the acteoside significantly reduced the escape latency of SAMP8 mice in training period.(2)In the experiment of autonomic activity, compared with the model group, the average speed and total distance of the low-dose acteoside group were significantly increased(t=15.0, 20.8, both P<0.05); the number of vertical movements and the average speed of the medium-dose acteoside group were significantly increased(t=15.8, 13.6, both P<0.05). The average speed and total distance of the high-dose acteoside group were remarkarbly increased(t=30.9, 29.7, both P<0.05), and the number of defecations in the mice of the lav-dose acteoside group, medium-dose acteoside group, high-dose acteoside group were((2.83±1.19)particles, (3.25±1.29)particles, (2.58±1.16)particles), they were obviously lower than that of the model group ((5.25±1.48) particles)(t=15.7, 20.1, 13.5, all P<0.01). (3)Simultaneously, the contents of ACh, NE and DA in the hippocampus and cortex of the model group were significantly lower than those in the normal group (hippocampus: t=10.3, 12.7, 13.2, all P<0.05; cortex: t=11.7, 10.5, 12.4, all P<0.05). Compared with the model group, the contents of ACh, NE, DA and 5-HT in the hippocampus and cortex of the low, medium and high doses of acteoside were significantly increased(hippocampus: t=31.4, 20.3, 10.7, 12.9, all P<0.05; cortex: t=33.7, 29.4, 14.5, 12.7, all P<0.05).@*Conclusion@#The acteoside can enhance the ability of spatial learning and memory of SAMP8 mice, and can regulate the depression and anxiety of animals.The mechanism may be related to the ACh content and the monoamine neurotransmitters increase in the brain.

Effects of Bushen Huoxue Acupuncture Method on Proteomics Expression in Amygdaloid Tissue of SAMP8 / 中国中医药信息杂志

Objective To observe the effects of Bushen Huoxue acupuncture method on amygdaloid protein expression in SAMP8; To explore the potential target protein for acupuncture treatment of Alzheimer disease (AD). Methods Thirty six-month-old male SAMP8 mice were randomly divided into acupuncture group and control group, 15 mice in each group. Acupuncture group selected Baihui (GV20), Shenshu (BL23), Geshu (BL17) and Xuehai (SP10) to intervene. The control group was given the same time to catch and stimulate. After 8 weeks, the amygdala was extracted and the differential expression protein spots were identified by proteomic techniques. Results Compared with control group, acupuncture group eventually identified 9 differential expression protein spots, of which 6 up-regulated and 3 down-regulated. According to the relevant information provided in the protein database, the main function of differential expression proteins involved in the mitochondrial energy metabolism, oxidative stress, and production of Aβ. Conclusion Bushen Huoxue acupuncture method can regulate multiple protein expressions in amygdala, suggesting that it may be through improving mitochondrial energy metabolism, oxidative stress, reducing production of Aβ to realize the potential therapeutic effects on AD.

Effect of electroacupuncture on Aβ and insulin degrading enzyme in the frontal cortex of senescence accelerated mouse/prone 8 / 中华行为医学与脑科学杂志

Objective To observe the effect of electroacupuncture at " Baihui"," Shenshu" and " Taixi" points on learning and memory,the expression of Aβ and insulin degrading enzyme(IDE) in the brain of SAMP8 mice.Methods Eight-month-old SAMR1 and SAMP8 mice were treated with electroacupuncture at " Baihui"," Shenshu" and "Taixi" points for eight consecutive weeks.The distribution and expression of Aβ and IDE in the brain prefrontal cortex of mouse were observed using immunohistochemistry and Western blot.Learning and memory function was assayed by Morris water maze(MWM).Results Compared with the normal control group,the brain prefrontal cortex of SAMP8 mice showed increased Aβ levels ((179.02± 15.11) ％),decreased IDE protein expression ((51.35 ± 14.94) ％).MWM showed the escape latency in SAMP8 mice was significantly longer than that in the control group (P＜0.05).Compared with the model group,Aβ levels in SAMP8 mice were markedly decreased in electroacupuncture group((119.72±9.21)％) and memantine-treated group ((116.84 ± 12.09) ％).IDE protein levels were increased by ((92.06 ± 8.05) ％) and ((83.84± 15.28) ％) after treatment with electroacupuncture and memantine relative to model group.MWM showed the escape latency was significantly shorter in SAMP8 mice treated with electroacupuncture and memantine than that in the model group(P＜0.05).Conclusion Electroacupuncture at " Baihui"," Shenshu" and "Taixi" points can inhibit Aβ levels,increase the expression of IDE and improve the learning and memory function of SAMP8 mice.

Fecal metabolome and gut microbiome alterations in a mouse model of senescence accelerated mouse prone 8 (SAMP8) / 中草药

Objective To investigate the alterations of fecal metabolites and intestinal flora during the aging in a mouse model of senescence accelerated mouse prone 8 (SAMP8). Methods The 1H-NMR metabonomics and metagenomics were applied to investigate the aging-related metabolic markers and intestinal flora, and Pearson correlation analysis was performed between metabolites and gut flora. Results Thirty-one endogenous metabolites were identified in the faeces of SAMP8 mice, of which 13 metabolites changed significantly compared with SAMR1 mice. Differential metabolites were mainly enriched in four metabolic pathways: phenylalanine, tyrosine and tryptophan biosynthesis; valine, leucine and isoleucine biosynthesis; phenylalanine metabolism; histidine metabolism. The results showed that the diversity of intestinal flora was significantly changed and the relative abundances of 10 kinds of intestinal flora were significantly changed in 10-month-old SAMP8 mice. Correlation analysis showed that Christensenellaceae was positively correlated with phenylalanine, histidine, valine, isoleucine, and uridylic acid; Dehalobacterium was negatively correlated with tyrosine, and Planococcaceae was negatively correlated with valine. Conclusion This paper reveals the changes of fecal metabolites and gut flora in SAMP8 mice, which provides experimental evidence for the study of aging progress and anti-aging actions of drugs.

Effects of Bushen Huoxue Acupuncture Method on Learning and Memory Abilities and Endogenous Neural Stem Cells Proliferation in Hippocampus of SAMP8 Mice / 中国中医药信息杂志

Objective To investigate the effects of Bushen Huoxue acupuncture method on the behavioral changes and the proliferation of endogenous neural stem cells (NSCs) in hippocampus of Alzheimer disease (AD) model mice. Methods 18 male SAMP8 mice, seven months old, were randomly divided into acupuncture group, non-acupoint control group, and model control group. And another age-matched 6 male SAMR1 mice were prepared as normal control group. Mice in acupuncture group were intervened by acupuncture method in the acupoints of "Shenshu", "Baihui", "Xuehai", and "Geshu". Mice in non-acupoint control group were treated by stimulating the fixed non-point under the bilateral rib, while mice in model control group and normal control group were raised without special treatment but administered the stimulation of catching with the same time and the same stimulus intensity. All treatrment lasted for 8 weeks. After the intervention, the learning and memory abilitities and brain hippocampus Brd U positive cells of all groups were detected. Results Compared with the nomal control group, mice in the model control group had longer escape latency and less time spent in former platform quadrant (P＜0.05); the number of Brd U positive cells decreased significantly (P＜0.05). Compared with the model control group and the non-acupoint control group, mice in the acupuncture group had shorter escape latency and more time spent in former platform quadrant (P＜0.05); the number of Brd U positive cells increased significantly (P＜0.05). Conclusion Bushen Huoxue acupuncture method can improve the learning and memory abilities of SAMP8 mice AD model by inducing the proliferation of hippocampal endogenous NSCs.

Effect of Acupuncture on Hippocampal Flotillin-1, NEP and Aβ42 Expressions in SAMP8 Mice / 上海针灸杂志

Objective To investigate the effect of acupuncture on behaviors and its mechanism of action in a SAMP8 mouse model of Alzheimer disease.Methods Sixty 6-month-old male SAMP8 mice were randomized to acupuncture,model and non--acupoint groups.Twenty male SAMP 1 mice of the same age constituted a normal control group.The acupuncture group received intervention by acupuncture at Shenshu,Baihui,Xuehai and Geshu.After eight weeks,a behavioral test was performed using the Morris water maze in every group of mice.The expressions of Flotillin-1,NEP and Aβ42 in mouse hippocampus were determined by western blot.Results The Morris water maze test showed that as compared with the model group,escape latency shortened significantly (P＜0.05) and the time spent in the former platform quadrant and the number of former platform position crossings increased (P＜0.05) in the acupuncture group of mice;escape latency,the time spent in the former platform quadrant and the number of former platform position crossings had no significant differences in the non-meridian-acupoint group of mice (all P＞0.05).As compared with the model group,Flotillin-1 expression decreased significantly in the acupuncture group (P＜0.05) but had no significant difference in the non-meridian-acupoint group (P＞0.05);NEP expression increased significantly in the acupuncture group (P＜0.05) but had no significant difference in the non-meridian-acupoint group (P＞0.05);Aβ42 expression decreased significantly in the acupuncture group (P＜ 0.05) but had no significant difference in the non-meridian-acupoint group (P＞0.05).Conclusions Acupuncture can markedly improve learning and memory abilities in a SAMP8 mouse model of AD and reduce Flotillin-1 content and upregulate NEP expression in SAMP8 mouse hippocampus to decrease Aβ42 expression,relieve neurotoxicity and produce a neuroprotective effect.

Tongluo Xingnao Effervescent Tablets ameliorates cognitive function of SAMP8 mice via Nampt/SIRT1/FOXO3 pathway / 中成药

AIM To investigate the effects of Tongluo Xingnao Effervescent Tablets (Chuanxiong Rhizoma,Angelicae sinensis Radix and Scutellariae Radix) on the learning and memory function in SAMP8 mice,and improvements in cognitive function.METHODS Morris water maze was used to evaluate the change of cognitive function in SAMP8 mice after they were treated with Tongluo Xingnao Effervescent Tablets for 60 d,and the activity of superoxide dismutase (SOD),the concentration of protein carbonyls (PC),the ratio of NAD +/NADH in brain tissue were detected by ELISA,the expression of Nampt,SIRT1 and FOXO3 in hippocampus were measured with immunohistochemical methods.RESULTS Tongluo Xingnao Effervescent Tablets shortened escape latency and obviously increased percentage of time in target quadrant in hidden platform test and increased the times entering the target quadrant in spatial probe test in SAMP8 mice.It evidently enhanced the activity of SOD,the ratio of NAD +/NADH and distinctly decreased the protein carbonyls level.Moreover,Tongluo Xingnao Effervescent Tablets could markedly up-regulate the expression of Nampt and SIRT1,but evidently down-regulate FOXO3 protein expression.CONCLUSION The experimental data show that Tongluo Xingnao Effervescent Tablets can enhance the cognitive function of SAMP8 through regulating Nampt/SIRT1/FOXO3 signal pathway.

Study of Waternut Herb Extract on Aβ SAMP8 Damage of Mouse Hippocampal neurons and the Protective Effect of Metabolomic Study / 现代生物医学进展

Objective:To study the metabolic mechanism of protective effect of waternut herb extract on primary Aβ SAMP8 damage of mouse hippocampal neurons by metabolic footprinting.Methods:MTT assay was used to determine the proliferation of primary hippocampal neurons in SAMP8 mice with Aβ damage,the effect for the first time on the basis of metabolic footprinting evaluation waternut herb extract.Focus on key metabolic pathways and related metabolic targets,mechanism of primary Aβ SAMP8 damage of mouse hippocampal neurons and pathogenesis of watemut herb extract.Results:MTT assay was used to measure the rate of cell proliferation.The results showed that the cell viability of the primary hippocampal neurons was significantly decreased in the Aβ SAMP8 mice.The study found that metabolic footprinting,compared with littermate wild-type mice,neuronal cell metabolism Aβ SAMP8 damage of mouse anomalies mainly concentrated in the metabolism of folic acid and taufine metabolism associated with nerve cells,by high-throughput mass spectrometric analysis and literature database retrieval to determine the 3 differential metabolites,respectively is L-disodoum alanine (L-Cysteic acid),dihydrofolate (Dihydrofolate),acid (Chorismate),the branch of small molecule metabolites through extract intervention after Amakusa callback trend obviously.Conclusion:the therapeutic effect of watemut herb extract on Aβ SAMP8 damage of mouse primary hippocampal neurons to a certain extent,3 biomarkers of this discovery may be a potential target of Aβ SAMP8 damage of mouse primary hippocampal neurons in the pathogenesis of waternut herb extract,given after these markers were callback trend in different degree,suggesting that watemut herb extract could regulate metabolism related enzymes and metabolic pathways to protect the purpose,to provide the experimental basis for the treatment of Alzheimer's disease watemut herb extract.

Effects of Qibao Meiran Oral Liquid on SOD activity and carbonyl protein content in brain tissue of mice / 中成药

AIM To observe the effects of Qibao Meiran Oral Liquid (Polygoni multiflori Radix Praeparata,Angelicae sinensis Radix,Psoraleae Fructus,etc.) on learning and memory function,hippocampus tissue pathological morphology,SOD activity and carbonyl protein content in SAMP8 mice.METHODS Twenty-seven SAMP8 mice were randomly and equally divided into model control group,donepezil hydrochloride group and Qibao Meiran Oral Liquid group.Another nine SAMR1 mice were selected as normal control group.Mice were given successive intragastric administration for 60 days.On the 56th day,the passive avoidance test was adopted,and the learning and memory capacities were determined after 5 d;The pathological morphology was observed by HE staining;ELISA assay was used to detect the activity of SOD and the content of carbonyl protein in brain tissue.RESULTS Compared with the model control group,the escape latency of mice in the Qibao Meiran Oral Liquid group was significantly prolonged,and the number of errors decreased significantly (P ＜0.01);the pathological morphology of hippocampus tissue was significantly improved;SOD activity increased significantly,and carbonyl protein content decreased significantly (P ＜ 0.01).CONCLUSION Qibao Meiran Oral Liquid can not only improve the learning and memory function of SAMP8 mice,but also reduce the degree of hippocampus tissue degenerative disease.

Effects of serums containing Buzhong Yiqi decoction with Astragalus Radix or Hedysari Radix on anti-immunosenescence in spleen lymphocytes of SAMP8 mice / 中国中药杂志

This paper was aimed to compare the effect of Buzhong Yiqi decoction containing Hedysari Radix or Astragali Radix on anti-immunosenescence effects in spleen lymphocytes of senescence accelerated mouse 8 (SAMP8). The effect of the serums on the proliferation of spleen T lymphocytes in SAMP8 mice induced by ConA was tested by MTT. The effect of the serums on the T lymphocytes subsets of SAMP8 mice was measured by flow cytometry. ELISA was used to detect the level of IL-2 and IFN-γ in the culture supernatants of spleen lymphocytes. The effect of the serums on the expression of CD28 mRNA in spleen T lymphocytes was detected by fluorescent quantitative PCR. Western blot was used to detect the expression of CD28 protein in spleen T lymphocytes of SAMP8 mice. Both the serums of Buzhong Yiqi decoctions containing Hedysari Radix or Astragali Radix improved the proliferation of T lymphocytes in SAMP8 mice. Both the serums had no obvious effect on the differentiation of spleen T lymphocytes'subsets in SAMP8 mice. Both the serums increased the content of IL-2 and INF-γ in the culture supernatants of spleen lymphocytes. And for the content of IL-2, the serum of Buzhong Yiqi decoction with Hedysari Radix was better（P<0.05）. Both the serums improved the expression of CD28 mRNA in spleen T lymphocytes of SAMP8 mice. And the effect of Hedysari Radix group was better than that of Astragalus Radix group（P<0.05）. Both the serums improved the expression of CD28 protein in spleen T lymphocytes of SAMP8 mice. The role of the serums containing Buzhong Yiqi decoction with Astragalus Radix and the decoction with Hedysari Radix in anti-immunosenescence was through the effect of the CD28. And the effect of Hedysari Radix group was better than that of Astragalus Radix group on improved the expression of CD28 mRNA in T lymphocytes of SAMP8 mice. Astragalus Radix and Hedysari Radix could swap in the aspect of anti-immunosenescence.

Study on histone H3 acetylation of BDNF gene promoter in SAMP8 mice / 中华行为医学与脑科学杂志

Objective To explore the role of histone H3 acetylation modification of brain derived neurotrophic factor (BDNF) in the pathogenesis of Alzheimer's disease (AD).Methods 2 months and 8 months SAMP8 mice were used as AD model.Morris water maze was used to detect the impairment of learning and memory.Western blot was used to detect BDNF protein expression in the hippocampus,and chromatin immunoprecipitation (CHIP) was applied to study the changes of histone H3 acetylation in different BDNF promoters.Results The results of water maze test showed that the time across the target quadrant in 8 months SAMP8 mice(0.9±0.4) was significant declined compared with that of 2 months SAMP8 mice(3.7 ± ±0.9) and 8 months SAMR1 mice (3.3±0.6)(all P＜0.05).Meanwhile,compared with 2 months SAMP8 mice ((23.9±4.0) s) and 8 months SAMR1 mice ((21.5± 2.3) s),target quadrant time in the 8 months SAMP8 mice((11.7±2.8) s) was also significantly reduced(both P＜0.05).The western blot showed the expression of BDNF in the hippocampus of 8 months SAMP8 mice was significantly decreased compared with that of 2 months SAMP8 mice and 8 months SAMR1 mice(P＜0.05).Lastly,CHIP assays showed that histone H3 acetylation of BDNF exon Ⅳ and Ⅵ in the hippocampus of 8 months SAMP8 mice were remarkably decreased(P＜0.05) compared with that of 2 months SAMP8 mice and 8 months SAMR1 mice.There was no significant change of histone H3 acetylation of BDNF exon Ⅰ and Ⅲ among all groups(P＞0.05).Conclusion Histone H3 acetylation of BDNF exon Ⅳ and Ⅵ is reduced during the development of AD,which may be the mechanism underlying the impairment of learning and memory in AD.