The Metabolomics Research Of Tongluoxingnao Effervescent Tablet In The Treatment Of Alzheimer’s Disease Based On Energy Metabolism Of Brain

ObjectiveThis project is to study how Tongluoxingnao effervescent tablet works in brain energy metbolism, brain energy metbolism and metbolic markers of AD’s rats by injected Aβ25-35in the rats’bilateral hippocampus. Then compare these reslut to summarize mechanism of Tongluoxingnao effervescent tablet.MethodWe use the morris water maze test to observe Tongluoxingnao effervescent tablet’s influence on AD’s rats by injected Aβ25-35in the rats’bilateral hippocampus capacity of learning and memory and use pathological test to observes changes of brain morphology. Then we observe energy metbolism by a-KGDH, PDH expression and average optical density in the hippocampal using immunohistochemistry method. We collected rat urine, applying UFLC-TOF MS method to analyze the metabolite of rat urine sample with statistical method such as PCA(Principal Component Analysis,PCA) and PLS-DA(Partial Least Squares-Discriminant Analysis,PLS-DA) to process data for Score plot and Loading plot. We acquired classified information from Score plot and Biomarkers from variables which contributed to classification to make clearly intervention effect of Tongluoxingnao effervescent tablet.Results1.AD rats’function of learning and memoryIn the training day2to5days, compared with the sham-operated group, escape latency of the model group are significantly extended(P<0.05); compared with the model group,the latency of the three doge group of Tongluoxingnao effervescent tablet are significantly shorter(P<0.05). After training, we began to do spatial probe test. The results show that the rats of model group’s time of staying in the first quadrant and platform is shorter than the sham-operated group(P<0.05) and the number of passing platform is less than the sham-operated group(P<0.05); the rats of Tongluoxingnao effervescent tablet groups’time of staying in the first quadrant and platform is longer than the model group(P<0.05) and the number of passing platform increse(P<0.05).2. brain pathology morphologyCompared with the sham-operated group, the rats of model group has the phenomenon of nerve cells decreased,neuronal apoptosis, hippocampus cell disorder and gliosis.Compared with the model group, the rats of Tongluoxingnao effervescent tablet groups are better in the phenomenon above.3. Expression of a-KGDH and PDHCompared with sham-operated group, positive immune response product of a-KGDH and PDH is much less in the model group. The expression of Tongluoxingnao effervescent tablet each dose groups are elevatory. The value of a-KGDH and PDH average optic density in the model group are lower than the sham-operated group significantly(P<0.05). High does and medium dose groups are significantly increased(P<0.05). Low dose group has a tendency to improve, but without statistical significance(P>0.05).4.Effects on metabonomicsScore plot shows each group is separated clearly. Compared with sham-operated group and control group, the model group is separated significantly. Tongluoxingnao effervescent tablet groups are closed to the control group, especially the middle dose group. The metbolic compounds of maximum contribution for classification is Indole-5,6-quinone, Cortisol,3-Mercaptolactate acid and4-Hydroxyphenylpyruvic acid. The abundance of these four metbolic compounds in Tongluoxingnao effervescent tablet groups are lower than model group(P>0.05).Conclusion1.Tongluoxingnao effervescent tablet can improve learning and memory ability of AD rats and protect the change of brain pathology morphologica induced by Aβ which suggests that it has effects on improving intelligence.2. Tongluoxingnao effervescent tablet can improve the activity of a-KGDH and PDH in hippocampus,which prompt that Tongluoxingnao effervescent tablet can improve brain energy metabolism by improving Efficiency of tricarboxylic acid cycle and glucose metabolism.3.Tongluoxingnao effervescent tablet can repair the system of dopamine, glucocorticoid and energy metabolism represented by biomarkers.