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Objective:To investigate the effect and mechanism of the Bushen Jianpi Kaixin formula on the learning and memory ability of Alzheimer's disease (AD) model rats.Method:A total of 72 SPF grade male SD rats were divided into control group, model group, Bushen group, Jianpi group, Kaixin group and Bushen Jianpi Kaixin group according to the random number table method ( n=12 in each group). The rats were intraperitoneally injected with D-galactose once a day for 6 weeks to replicate the model of AD.And the rats in different medication groups were given corresponding administration (Bushen formula: gavage 3.60 g·kg -1·d -1, Jianpi formula: gavage 4.05 g·kg -1·d -1, Kaixin formula: gavage 2.34 g·kg -1·d -1, Bushen Jianpi Kaixin formula: gavage 9.99 g·kg -1·d -1), while rats in control group and model group were treated with equal volume of 0.9% sodium chloride solution once a day for 28 days.The learning and memory ability was tested by Morris water maze.The expressions of phosphoinositide 3-kinase(PI3K), protein kinase B(Akt) and mammalian target of rapamycin(mTOR) in cerebral cortical tissues were detected by immunohistochemistry.The relative mRNA levels of p62 and Beclin in brain cortical tissue were detected by RT-PCR.SPSS 25.0 software was used for data processing, one-way ANOVA was used for inter group comparisons, and LSD test was used for further pairwise comparisons. Results:Morris water maze results showed statistically significant differences in escape latency and the times of crossing platform among the six groups ( F=368.10, 47.43, both P<0.01). The escape latency of Bushen Jianpi Kaixin group((29.30±1.64) s) was shorter than that of model group((55.58±3.23) s) ( P<0.01), the times of crossing platform ((5.17±0.72) times) in Bushen Jianpi Kaixin group was higher than that of model group (1.50±0.52)time, P<0.01). Compared with the Bushen Jianpi Kaixin group, the escape latencies of Bushen group, Jianpi group and Kaixin group were longer (all P<0.01), the times of crossing platform in Bushen group was lower ( P<0.01). Immunohistochemical results showed statistically significant differences in the positive protein expression of PI3K, Akt, and mTOR proteins among the six groups ( F=68.52, 22.22, 31.52, all P<0.01). Compared with the model group, the levels of positive protein of PI3K ((0.47±0.15), (0.57±0.12)), Akt ((0.31±0.02), (0.38±0.02)), and mTOR ((0.22±0.18), (0.28±0.11)) in Bushen Jianpi Kaixin group were less (all P<0.01). Compared with the Bushen Jianpi Kaixin group, the levels of positive protein of PI3K and mTOR of Bushen group, Jianpi group and Kaixin group were higher (all P<0.01). RT-PCR results showed statistically significant differences in the relative mRNA levels of Beclin and p62 among all the groups ( F=8.79, 21.01, both P<0.01). The relative mRNA level of Beclin in Bushen Jianpi Kaixin group was higher than that of the model group ((0.97±0.07), (0.64±0.12)), and the relative mRNA level of p62 of Bushen Jianpi Kaixin group was less than that of model group((0.98±0.16), (1.16±0.24))(both P<0.01). The relative mRNA levels of p62 in Bushen group, Jianpi group and Kaixin group were higer than those of Bushen Jianpi Kaixin group (all P<0.05). Conclusion:Bushen Jianpi Kaixin formula can improve cognitive impairment and learning and memory ability in AD model rats.The mechanism may be related to the regulation of PI3K/Akt/mTOR autophagy pathway.The combination prescription is better than the split prescription.
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This study successfully established in v itro model of Alzheimerˊs disease ( AD ) in order to investigate effects and mechanisms on AD by Bushen-Huatan-Yizhi Recipe (BSHTYZR, replenishing kidney-essence and removing phlegm to benefit wisdom ) . The PC-12 cells affected by Aβ1-42 incubated were produced for neuron cell model of AD . Then , they were intervened by the serum containing BSHTYZR , which were compared to the serum containing indomethacin . The cell morphology was observed . The neuronal survival was assessed by counting MTT assay . The expression of APP mRNA was detected with RT-PCR . And the protective effect of BSHTYZR on neuronal cells and neurotoxic effects of anti-Aβ was observed . The results showed that 2 . 5 mmol/L Aβ1-42 can significantly decrease the cell viability . The RT-PCR was used on semi-quantitative analysis for expression of APPmRNA. It showed that expression of APP mRNA was increased in each group, and the BSHTYZR group can reduce the expression significantly ( P < 0 . 01 ) . It was concluded that the BSHTYZR method can protect nerve cells , suppress the expression of APP mRNA , and reduce the toxic effects .
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Objective To observe the effects of wild jujube seed decoction(WJSD) on ultrastructure and astrocytes expression in the brain cortex of old model rats with blood-deficiency and Yin and sleep deprivation(SD)after SD. Methods Ultrastructural changes in cortical parts of the experiment rat were observed by transmission electron microscopy , immunohistochemical staining was used to detect star-shaped glial cells marker-glial fibrillary acidic protein(GFAP) expression after SD induced by made-self multiple platform method (MMPM), senescence induced by D-galactose, and Yin and blood-deficiency induced by cyclophosphamide and hydrocortisonum. Results Compared to environmental control group((9.8 ± 2.5), (0.11 ± 0.02) ) , syndrome model group (( 20.4 ±4.4), (0.20 ±0.011) ) rats had obvious ultrastructural changes and stronger expression of GFAP in cortical parts (t = 7.63,3.18, P ＜ 0. 01), while those of W J SD of high and low dose group ((14.4 ± 3.9), (15.5 ± 6.4),(0.14 ±0.02)(0.14±0.02)) rats showed weaker(t=6.32,5.24,2.31,2.45, P＜0.05). Conclusion WJSD of large and small lose group could improve the rats neurons pathological changes,WJSD downward adjusting the expression of GFAP may be one of the mechanisms of treatment on insomnia with blood-deficiency and Yin in the old.
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Objective:To observe the effect of replenishing the kidney and eliminating phlegm therapy on calciumlcalmodulin-dependent protein kinase Ⅱ(CaMKⅡ-?) activities of AD rat model,and explore the potential mechanism.Methods:AD model was established by injection of ibotenic acid(IBO) and 25-35 ?-amyloid peptide(A?25-35) into basal nucleus of Meynert.After injecting for two weeks,Bushen Huatan Decoction was administered by oral gavage of high,middle and low dosages.Huperzine A-Zhulin Antun tablets served as control drug.Normal group,model group and blank group were administered with isovolumetric saline water.After treating for four weeks,hybridization in situ was used to observe the change of CaMK Ⅱ in hippocamp.Results:The expression of CaMKⅡ-? in model group was more than that in normal group(P
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Objective: To investigate the effect of Modified Wendan Decoction(MWD) on expression of p-JNK and p-c-Jun in cellular model of AD.Methods: NG108-15 cells were exposured to 5?mol/L A?25-35 for 24h to establish the cell model.After treating with contained-MWD serum,cells morphous were observed,survival rate was tested by MTT and cell apoptosis by FCM,the expression of p-JNK and p-c-Jun was measured by method of immuocytochemistry.Results: MWD obviously reduced the expression of p-JNK and p-c-Jun and cell's apoptotic rate.Compared with the model group,the expression of p-JNK and p-c-jun in the SP600125 group and the group treated with contained-MWD serum decreased significantly(P