Function And Mechanism Of RhoA And Cytoskeleton Actin In Porcine Pseudorabies Study On The Role And Virus Infection

Pseudorabies(PR),also known as porcine herpes,Aujeszky’s disease(AD),or pruritus,is caused by the Aujeszky’s disease virus(ADV),pseudorabies virus(PRV),or porcine herpesvirus type I(Su HV-1),and is one of the most harmful infectious diseases worldwide,causing huge economic losses to the pig industry.Cytoskeleton mainly includes microfilament(MF),microtubule(MT)and intermediate filaments(IF).They form protein fiber network system in host cells,which not only plays an important role in maintaining cell morphology and maintaining the order of internal structure of cells,but also participates in many important life activities,such as cell movement,material transport and information transmission.In addition,it can also help some viruses to complete a series of life activities such as adsorption,invasion,intracellular transport,replication,assembly and release,help viruses escape from the immune system,and enable viruses to survive and proliferate.As a member of the small molecule Rho GTPase family,RhoA can affect the morphological changes of microfilaments by regulating downstream related proteins,thus participating in various cellular physiological and biochemical functions.Studies have shown that Rho GTPase is involved in viral infection,but whether RhoA is involved in PRV infection and its molecular mechanism are still unclear.This paper mainly studies the role and molecular mechanism of RhoA and microfilament in PRV infection.Through chemical drug treatment and gene knockdown / overexpression strategies,through morphological observation,flow cytometry,immunofluorescence,western blotting,RT-q PCR and viral titer determination were used to analyze the function of RhoA in PRV infection at the cellular,protein and molecular levels,and to further analyze the molecular mechanism of RhoA regulating PRV infection.The results showed that RhoA inhibitor Rhosin promoted the proliferation of PRV,while RhoA agonist Narciclasine inhibited the proliferation of PRV.Knockdown of RhoA promoted PRV proliferation,while overexpression of RhoA inhibited PRV infection.Secondly,RhoA inhibits PRV replication by inducing cytoskeletal microfilament rearrangement.Finally,we demonstrated that RhoA regulates PRV infection by limiting virus adsorption and entry through time addition experiments.In summary,the results show that RhoA is involved in PRV infection by regulating cytoskeleton microfilament rearrangement,and inhibits PRV infection by limiting virus adsorption and entry.These studies on the role and mechanism of host cytokines in PRV infection can not only deepen our understanding of the pathogenesis of PRV,but also provide a theoretical basis for vaccine development and PR prevention and control.