| Objective Parkinson’s disease (PD) is common in the elderly degenerativedisease of central nervous system, it affects the quality of life above60years ofage in the elderly population by2%.PD has the character of onset occult, noobvious early symptoms, serious disability, continued progress, cannot bereversed and not curable in the present. The current research is focus on initialstage of the disease. Because it is almost impossible to make the diagnosis inpatients with early clinical stage at present and research leading to directlyhuman PD pre-clinical risk early warning mechanism can not be carried out.There is an important protein in the pathogenesis of PD named α-syn, itsabnormal aggregation is often considered as the main reason leading to neuronaldegeneration. There are reports that the progression of PD could be due to α-syndiffusion from gastrointestinal nervous plexus to the nervous system,involvement of medulla oblongata, with the increase in the number of neuronsinjury and appear clinical symptoms. So this thesis choose the most similar tohuman physiological primate animal cynomolgus monkey to research on animalmodel of PD and the construction of alpha synuclein index system. The researchis of great significance to solve the PD pre-clinical pathological development evaluation and classification evaluation.Methods1. Choose9healthy cynomolgus monkey with different age as theanimal model of natural aging,3young monkeys (4years old),3middle agedmonkey (10to11years old),3old monkey (15to16years old). Test theexpression ofα-synand n-syn in different ages of colon in cynomolgusmonkeys by immunohistochemical method. Observe the colocalization of-synand n-syn by double immunofluorescence.2. Choose9healthy cynomolgusmonkey with different age as the animal model of MPTP(1-methyl-4-phenyl-1.2.3.6-tetrahydrophridine),3young monkeys (4years old),3middle aged monkey (10to11years old),3old monkey (15to16years old).Test the expression ofα-synand n-syn in different ages of colon in cynomolgusmonkeys by immunohistochemical method. Observe the colocalization of-synand n-syn by double immunofluorescence.Results1. Double immunofluorescence staining results confirmed that there iscolocalization ofα-synand n-syn in the colon in natural aging model ofcynomolgus monkeys. Immunohistochemical analysis revealed thatα-synandn-syn expressed in the colon in natural aging model of cynomolgus monkeyswith different age. However,α-synand n-syn showed the highest expressionlevel in the model of old cynomolgus monkeys,in the opposite, the lowest levelin the model of young cynomolgus monkeys.2. Double immunofluorescencestaining results confirmed that there is colocalization ofα-synand n-syn in thecolon in MPTP model of cynomolgus monkeys. Immunohistochemical analysisrevealed thatα-synand n-syn expressed in the colon in MPTP model ofcynomolgus monkeys with different age.However,α-synand n-syn showed thehighest expression level in MPTP model of old cynomolgus monkeys,in theopposite, the lowest level in MPTP model of young cynomolgus monkeys. Conclusion1. In this experiment, immunohistochemistry results of naturalaging model of cynomolgus monkeys showed that with age ageingα-synandn-syn aggregation of the protein expression increased, fluorescent doublelabeling showed the coexistence ofα-synand n-syn. The result confirmed thatthe natural aging has positive relation with the oxidative modification of-syn,it can increase the expression of toxicity polymer with n-syn.2. In thisexperiment, immunohistochemistry results of MPTP model of cynomolgusmonkeys showed that with age ageingα-synand n-syn aggregation of theprotein expression increased, fluorescent double labeling showed thecoexistence ofα-synand n-syn. The result indicate that aging of promptedα-syncontent has increased the toxicity of MPTP, co-promote the modificationreaction ofα-synnitration, eventually lead to colon tissue of aged monkeysMPTP intestinal myenteric plexus with high expression of n-syn.3. Theseexperiments were carried out to verify the theory ofα-synin PD in thepathological process that might start from the peripheral nervous system.In-depth studies to continue with PD sensitive brain regions on the mechanismare needed. |
PDF Full Text Request