A Small Study On The Role Of Hsp60 And Rab2 Of ME49 Strain Of Toxoplasma Gondii

Toxoplasma gondii(T.gondii)is a human zoonotic parasite,which is seriously harmful to human beings.The pathogens of infection include cysts,oocysts and tachyzoites.Tachygonites depend on parasitism in cells and have higher requirements for their own living environment.Therefore,effective drugs for T.gondii are mainly acting on the stage of T.gondii currently.T.gondii infection is mainly due to the successful invasion of all cells containing nucleus by T.gondii.After invasion,it can successfully evade host immune defense by self disguise.T.gondii infected animal is mainly to show clinical symptoms when the body’s immunity is low.The main symptoms are Toxoplasma encephalitis(TE)and other brain diseases,even cause paralysis due to the neurotropic characteristics of the worms.But for brain diseases,calcium overload is considered as a common pathway of neuronal damage in many kinds of brain injury diseases.In general,the calcium permeability of neurons is increased by some reasons.The calcium channels that lead to calcium overload in neurons are expressed differently in different parts of the neuron,and there are also interactions between different calcium channels.In previous study,We selected differentially expressed proteins Drebrin(Dbn1)as a bait protein by Co-immunoprecipitation and Pull-down,the protein interact with Dbn1,exosome protein accounted for 4,the difference was verified by qPCR more significant included Ras protein(Rab2),heat shock protein(Hsp60).In view of the structure of exosome and characteristics of blood-brain barrier,it is necessary to further study the mechanism of interaction between host and parasites in Toxoplasma encephalopathy.In this study,the PCDH-CMV-EF1a-GFP-puro lentivirus vector was used to prepare the lentivirus recombinant plasmid CMV-Hsp60 and CMV-Rab2 to prepare the viral brain injection mice to observe the behavior of mice through animal behavior experiments,and introduced Hsp60 and Rab2 with the HA tag by pcDNA3.1(+)eukaryotic expression in mouse neuroblastoma cells(N2a)and Western blotting(WB)was used to detect the expression levels of Dbn1,Bcl-2 and LC3 I / II.The experimental results showed that the behavior of the mice infected virus that containing Rab2 and Hsp60 had changed compared with the control group,and the N2 a cells transfected with the eukaryotic expression vector of the target gene had obvious apoptosis after the transfection,compared with the control group.This indicates that Rab2 and Hsp60 have effect on animal behavior,and may promote the occurrence of apoptosis.The increase of the infection rate in the cells treated with rapamycin may mean that the occurrence of autophagy promotes the invasion and proliferation of the insect body at the early time,and these studies provide the basis for the prevention and treatment of Toxoplasma.