The Influence Of Rotenone Toxicity On PrP Knockout Mouse And Its Mechanism Research

This thesis mainly focus on the toxicity of Rotenone on pure line wild-type mice and Prion protein(PrP)knockout homozygotes(PrP-/-)transgenic mice.Investigating that whether PrP can be protective to Rotenone-induced acute toxicity and effects of Parkinson’s disease.Rotenone is an odourless,crystalline ketonic chemical compound used as a broad-spectrum insecticide.It occurs naturally in the seeds and stems of several plants,such as the jicama vine plant;and the roots of several members of Fabaceae.The toxicity of rotenone can inhibit the mitochondrial electron transport chain of cell,which will reduce ATP levels in vivo,and finally cut down the energy supply of animals,made them paralysis and death.Some studies have shown that rotenone can lead the formation of neurons in the substantia nigra a-synuclein-positive aggregates,but the exact mechanism is still not clear.There are large numbers of animal experiments using rotenone to establish the animal model of Parkinson’s disease.Some existing research shows that the humoral immune response against PrP can prevent the prion infection.In vitro experiments,identifying PrP antibodies or fragment of antigen binding(Fab),could prevent prion infection.As a neurodegenerative disease--Parkinson’s disease,so we infer that,the pure line wild-type mice have stronger ability rotenone acute and chronic toxicity than the PrP knockdown homozygotes transgenic mice.To explore the above inference,we conducted the following experiments:(1)acute intraperitoneal injection of rotenone,comparing their behavior status and survival curves;(2)acute intraperitoneal injection of rotenone,killed them in different time after injection,using TH cell immunohistochemistry and physiological and biochemical detection of their brain and other tissues,explore the difference between two types of mice in the face of the acute toxicity of rotenone;(3)Using nasal drip of rotenone,killed the mice after a period of time and detect two types of mice’ dopa钟 e cell survival situation in brain tissue,particularly the olfactory bulb part.In this study,first we tested the PrP knockout mice’ genetic to ensure it is PrP knockout homozygous.Tasted by PCR tests that they were homozygous knockout(PrP-/-)transgenic mice,and PrP were not expressed in its various tissues and organs,WT mice were normal expressed PrP in its various organs.Toxicity testing were all on animal levels,which found during the acute intraperitoneal injection of rotenone,a short time after ad分钟 istration of two types of mice were choking convulsion,inbred wild-type mice died within a short time,PrP knockout homozygotes(PrP-/-)transgenic mice survival situation is more favorable.The death rate of two type’s mice’cell in different organs did not appear significantly.After testing,the intraperitoneal ad 分钟 istration group,two types of mouse brain tissue NADPH,NADP,NAD content did not differ significantly(P>0.05).Nasal groups were not significantly different types of mouse content NADP and NAD(P>0.05).The experiment also shows that PrP knockout mice and WT mice have clear difference rotenone chronic toxicity.The TH positive cells in substantia nigra of PrP-/-mice lost more heavily than WT mice,which indicate that PrP can prevent the dopa 分钟 ergic cell death induced by rotenone.