| Alzheimer's disease(AD)is a neurodegenerative disease that affects people over the age of 65.AD is characterized by senile plaques,neurofibrillary tangles,and extensive hippocampal neurons.Modern medicine divides senile dementia into AD,vascular dementia,mixed dementia and other dementias.AD and vascular dementia are the two most common.AD,an a chronic disease,is difficult to reverse once it occurs.AD process is relatively complex,more focused on several hypothesis:A?neurotoxicity hypothesis,hypothesis of excessive Tau protein phosphorylation,gene mutation hypothesis,cholinergic neurons damaged hypothesis,hypothesis of excitatory amino acids toxicity,disorder of nerve cell calcium balance,high cholesterol hypotheses and so on.The above possible pathogenesis shows the complexity of AD.In the process of looking for the treatment of AD drugs,we should try to analyze the treatment effect of drugs on the treatment objects from multiple perspectives,and then find and look for the possible intervention ways and means of drugs.Huanglian jiedu decoction(HLJDD)is the representative prescription for clearing away heat and detoxifying.It is mainly made up of scutellaria baicalensis,Coptis chinensis,golden cypress and gardenia in a ratio of 3:2:2:3.Under the guidance of the theory of traditional Chinese medicine,the above four drugs are bitter and cold,easy to hurt the spleen and stomach,not easy to overdose or take for a long time.On the basis of the previous experiments of our study,AD rats induced by A?25-35 and Ibotenic acid combined with HLJDD at a clinical dose for one week showed a significant decline in their living conditions(including yellowing).Therefore,to explore the mechanism of HLJDD in Tg-APP/PS1 mice,the dose was reduced to 1/20 and 1/10 times of the equivalent dose of HLJDD in mice.1?Effects of HLJDD on the behavior of Morris water maze in Tg-APP/PS1 miceIn the navigation test:In the first day of training,the model group showed significant difference(P<0.05)compared to the control group,but the others are not.With the increase of training times,the escape latency of each group of mice gradually decreased,the gap between the control group and the model group was more and more significant,and the advantage of the treatment group was obvious.After the end of the third day of training,the escape latency of the model group was much greater than that of the control group(P<0.001),but the others was significantly lower than that of the model group.The spatial probe test:According to the motion trajectory diagram,the model group has no purpose as the control group,the spatial memory capacity of Tg-APP/PS1 mice was improved dramatically under the action of H-L,H-H,and Don but not Ber.From the navigation test and the spatial probe test,the order in which the drugs restored the cognition of the Tg-APP/PS1 mice was H-L>H-H>Don>Ber.2?Effects of HLJDD on the A?,Tau,and the oxidative stress state in Tg-APP/PS1 miceSignificant accumulations of A? were observed in both cortex and hippocampus of Tg-APP/PS1 mice.HLJDD could alleviated this deposition of A? in brain,and it was worth noting that the effect of H-L evaluated by the sediment and size was superior to that of H-H,Ber,and Don.Meanwhile,TNFs were observed in the hippocampus but not cortex of mice.Overall,the TNFs were not obvious,but it is still found that the degree of lesion in Mod was higher than that in other groups.Compared to the con group,the contents of SOD in brain dramatically decreased in the Mod group(P<0.001).After administration of drugs,the brain concentration of SOD was increased significantly under the action of H-L and Ber,and the levels of MDA was declined by H-L and increased due to the intervention of Don,relative to the Mod group.3?Effects of HLJDD on the central neurotransmitters in Tg-APP/PSl transgenic miceCitrulline,methionine and asparaginate as the difference(P<0.01,P<0.01,P<0.05 respectively)were screened out from Tg-APP/PS1 mice relative to normal mice.However,the rest contributes less to pathology of Tg-APP/PS1 mice in present study.In this study,we performed a detailed analysis to determine the neuroprotective effect of HLJDD on Tg-APP/PS1 mice with double dose of 5g/kg/d and lOg/kg/d.We observed both H-L and H-H significantly increased the contents of brain PHE,arginine,proline,tyrosine,Trp,5-HT,L-leucine,L-isoleucine,threonine,and asparaginate and markedly decreased the levels of L-cysteine compared to Mod.Additionally,Glu,choline,and GABA raised significantly in H-H and urea declined dominantly in H-L relative to Mod.4?Effects of HLJDD on the central and peripheral inflammation in Tg-APP/PSl miceCompared with Con,3 and 4 differential cytokines were found in the central and peripheral systems of Tg-APP/PS1 mice respectively.Among them,IFN-?(declined in Mod;P<0.05)and IL-6(raised in Mod;P<0.05)were the shared differences in both the central and peripherical system;IL-12p70(declined in Mod;P<0.01)was unique to the central system;IL-13(declined in Mod;P<0.01)and MCP-1(raised in Mod;P<0.01)were the special difference of the peripherical system.After drugs,the cytokines in the central and peripheral areas have changed to varying degrees.H-L and H-H could ameliorate the contents of IFN-?,IL-1?,IL-1?,IL-4,IL-6,IL-10 and MIP-2 of the central system and reversed the levels of IFN-y,IL-la,IL-1?,IL-6,IL-12p70,IL-13,MCP-1 and MCP-2 of the peripherical system to some degree.5?Effects of HLJDD on the central and peripheral lipids metabolism in Tg-APP/PS1 miceTarget lipids:Subsequently,40 pathological lipids biomarkers in brain tissue were identified.Comparing with the normal mice,the levels of 17/21 PCs,5/5 PEs,3/3 GlcCers,4/4 Cers,and 5/7 SMs went decreased in Tg-APP/PS1 mice brain and the remains increased.Comparing with the model group,18/21 PCs,3/5 PEs,1/3 GlcCers,3/4 Cers,and 2/7 SMs were reversed to different degree in both low and high dose of HLJDD.Simultaneously,13 pathological lipids biomarkers in serum,including 11 PCs,1 PE,and 1 GlcCerds,were screened.Almost all pathological lipids biomarkers,except for PC(20:4/18:0),dropped in the model group and were reversed in both H-L and H-H.Consequently,the lipids metabolites in the central and peripheral regions were altered in Tg-APP/PS1 mice,particularly in central system,and H-L and H-H could alleviate this lesion.Polyunsaturated fatty acids:The overall pattern of results showed no significant differences of brain PUFA in Tg-APP/PS1 mice relative to wild mice,but the serum contents of omega-6 acids(LA,AA,and GLLA)and omega-9 acid(OLA)were significantly reduced.After administration of drugs for 4 months,H-L and H-H significantly increased the brain levels of AA,DHA,EPA,LA,and OLA and declined the contents of GLA,but the measured fatty acids of the serum differ very little in H-L and H-H compared to MOD.Simultaneously,the regulatory effect of Ber and Don on the central and peripherical PUFA was similar to that of H-L and H-H,but it was clear that Ber's regulation of central PUFA was far less than that of H-L and H-H.6?Effects of HLJDD on the serum bile acid metabolism in Tg-APP/PS1 miceBased on LC-QQQ-MS technology,the content of bile acid in serum of Tg-APP/PS1 mice was determined.Through the analysis of the results,it was found that Tg-APP/PS Imice had abnormal peripheral blood clear bile acid metabolism.Compared with the normal group,DCA(P<0.001),T-?-MCA(P<0.05)and THDCA(P<0.05)in serum of Tg-APP/PS1 mice were significantly increased.With the intervention of drugs,bile acid levels in serum of both Tg-APP/PS1mice were adjusted to different degrees.H-1 significantly down-regulated the levels of DCA(P<0.05),T-a-MCA(P<0.05)and THDCA(P<0.05)in the serum of AD mice.H-H can significantly reduce the level of THDCA in serum(P<0.05).Berberine significantly down-regulated T-a-MCA(P<0.05)and THDCA(P<0.05)in AD mice.However,allianz had a significant downregulation effect on THDCA(P<0.05).7?Effects of HLJDD on Gut Microbiota in Tg-APP/PSl miceIn this chapter,16 sRNA high-throughput sequencing technology was used to explore the regulation of HLJDD on intestinal flora of the Tg-APP/PS 1 mice.According to the PCA score,it can be intuitively found that all tested samples are mainly divided into three camps.Among them,the samples of Con group and Mod group are close to each other,but can be obviously separated,indicating that the intestinal microbial structure of Tg-APP/PS1 mice is out of order.While the regulating effects of H-L,H-H and Don on the intestinal flora of Tg-APP/PS1 mice were similar,the regulating mechanism of berberine on the intestinal flora of Tg-APP/PS1 mice was different from that of H-L,H-H and Don.Compared with the normal group,the relative abundance of bacteroidetes and actinomycetes in the model group decreased,while that of proteobacteria and firmicutes increased.After administration,in comparision with the model group,the relative abundance of bacteroidetes decreased in the H-L group,the H-H group and the Don group,and increased in the Ber group.The relative abundance of firmicutes decreased in the h-h group and the Ber group,and increased in theH-L and Don groups.The relative abundance of proteobacteria and actinomycetes was increased in the H-L group,H-H group,Ber group and Don group.Histogram analysis of sample OTU abundance was conducted at the level of family and genus respectively.The existing laws can be intuitively found from the different color distributions of different bacteria in different groups,that is,the expression forms of H-L,H-H and Don groups are similar,while the structural changes of the flora in Ber group at the level of family are obviously different from those in other groups. |
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