Compound Danshen Alzheimer Animals And Mechanism Study

Alzheimer's disease (AD) is a neurodegenerative disorder. The clinical characteristic of AD is recessive invasion. The memory impairment, cognition disturbance, behavior abnormality, and communication disorders will gradually appear. The patients will gradually lost their ability of independent living, and the quality of their life will decrease dramatically, which furthers the burden for their family and the whole society. Even worse, with the elderly population increase in recent years, the incidence of AD goes up. Therefore, the governments and the research institutions need to make great efforts to explore this disease. In China, besides the western medicine, we have an unique medical system called Chinese medicine that was clinically tested for thousands of years. The Chinese medicine provides a plenty of effective therapeutics that deserve further investigation. It is really our mission and responsibility to explore and take advantage of the unique system to get cures for such a neurodegeneration disease.In Chinese medicine, the therapeutic principle of dementia related disease is to improve blood circulation and remove blood stasis. Based on this theory, we developed the research idea of this study by choosing the medicine of Fufangdanshen (FFDS), which is a classic formula used for promoting blood circulation and removing blood stasis. FFDS is simply compose of danshen root, notoginseng radix, and borneo camphor. In the research, we investigated the effects of FFDS on several animal models of dementia. The objective of this study is to test whether FFDS is effective for Alzherimer's disease.1 Effects of FFDS on the brain function of D-gal and NaNO₂ trigered aging miceIn this study, D-gal and NaNO₂ were injected i.p. once a day consecutively for 53 days to trigger the aging process of mice. FFDS was administrated once a day by p.o. during the whole period of experiment. We then observed the effects of FFDS on animals'ability of learning and memory. The biochemical changes and the histo-pathology of the brain were also investigated.1.1 Effects of FFDS on learning and memory ability of D-gal and NaNO₂ treated aging miceThe results showed that the learning and memory ability of D-gal and NaNO₂ injected mice decreased after 53 d, with markedly increased numbers of entering to blind-ending and lengthened latency. Treatment with FFDS improved the learning and memory ability of triggered mice. Both the training and test time are significantly shorten by FFDS treatment. The mistake numbers also decreased after FFDS treatment. The results indicate that FFDS had a therapeutic potential for AD. Based on our results, it seemed that product B was the most effective ones.1.2 Effects of FFDS on the neuronal cells death in hippocampus and cerebral cortex of D-gal and NaNO₂ treated miceAfter 63 d injection of D-gal and NaNO₂, lots of neurons in hippocampus and cerebral cortex were damaged, which was confirmed in nissl stain by methylene blue. Treatment with FFDS dramatically improved the situation of damage neurons in hippocampus and cerebral cortex.1.3 Effects of FFDS on brain biochemistry metabolism of D-gal and NaNO₂ treated miceAfter 60 d injection of D-gal and NaNO₂ , the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the brain decreased, the same time malondialdehyde (MDA) increased significantly. All FFDS groups could significantly relieve free radicals damage by enhancing the activity of SOD and GSH-Px and reducing the content of MDA.1.4 Effects of FFDS on the activity of choline acetyl transferase and acetyl cholinesterase in hippocampus and cerebral cortex of D-gal and NaNO₂ treated miceThe results showed that 60 days injection of D-gal and NaNO₂ resulted in an increase of the activity of AChE and a decrease of ChAT activity in the brain. Administration with FFDS did not change activity of AChE, however, the activity of ChAT both in cerebral cortex and in hippocampus was significantly enhanced by FFDS.2 Effects of FFDS on amyloid beta peptide injected ratsThe main pathology of AD closely correlated with toxicity of amyloid beta peptide (Aβ). In the study we deployed a classic animal model of AD, a model of injecting aggregative Aβ_1-40 into hippocampus of rats to induce a dementia symptom. 2.1 Effects of FFDS on learning and memory ability of amyloid beta peptide injected ratsWe injected Aβ_1-40 into hippocampus of adult Sprague-Dawley rats, and applied Morris maze to test learning and memory ability in all groups of rats. In spatial discrimination experiment, the rats showed a remarkable time prolonging in platform searching, with a significant increase in swimming distance after Aβ_1-40 injection. Administration of FFDS could significantly alleviated the impairment in spatial discrimination caused by Aβ_1-40, which was proven by shorten the swimming time and distances before the rats found the platform.2.2 Effects of FFDS on neuronal cells death in hippocampus and cerebral cortex of amyloid beta peptide injected ratsWe injected Aβ_1-40 into hippocampus of adult Sprague-Dawley rats, and then regularly processed the brain sections for histochemistry. Methylene blue was used as the stain to assess the the condition of neurons in hippocampus and cerebral cortex. The results showed the injection of Aβ_1-40 caused the disappearance of methylene blue-stained neurons in CA1 of the hippocampus. Treatment of animals with FFDS could significantly prevented CA1 neural cells loss. Cerebral cortex neurons had no obvious change before and after Aβ_1-40 injection.2.3 Effects of FFDS on the activity of choline acetyl transferase and acetyl cholinesterase in hippocampus and cerebral cortex of amyloid beta peptide injected ratsWe injected Aβ_1-40 into hippocampus of adult Sprague-Dawley rats, and employed biochemistry in order to investigate the effects of FFDS on activity of AChE and ChAT in rats brain. The results showed the injection of Aβ_1-40 caused a significant decline of ChAT activity, however, AChE activity had no obvious change. Administration of FFDS could remarkably improved the activity of ChAT in hippocampus; FFDS had no effect on AChE activity.2.4 Effects of FFDS on the formation of neural plaque in hippocampus of amyloid beta peptide injected ratsWe injected Aβ_1-40 into hippocampus of adult Sprague-Dawley rats, and observed the deposition of senile plaques using thioflavin-S stain. The results showed senile plaques increased in hippocampus after Aβ_1-40 injection, and FFDS at high dose could obviously conversed the situation. 2.5 Effects of FFDS onβ-secretase activity in hippocampus of amyloid beta peptide injected ratsWe injected Aβ_1-40 into hippocampus of adult Sprague-Dawley rats, and applied immunohistochemical method to observedβ-secretase (BACE) activity in hippocampus. The results showed BACE activity in model group animals had an increasing tendency, FFDS could slightly decreased its activity, however, there is no significant difference between groups.In this study we showed that FFDS could significantly improve experimental AD animals'learning and memory ability; improved the situation of neurons impairment; enhanced the activity of SOD and GSH-Px; reduced the content of MDA; increased ChAT activity in hippocampus and cerebral cortex. All the results above indicated that FFDS could be a promising candidate for AD therapy.