Effects Of Changes In Extra And Intracellular Gelsolin On Polymorphonuclear Neutrophil Chemotaxis

Part ⅠObjective:To investigate the effects of gelsolin (GSN) itself on rat polymorphonuclear neutrophil (PMN) chemotaxis.Methods:PMN were isolated from rat venous blood. PMN suspension was seeded to the upper chamber of transwell culture system and recombinant GSN was added to the lower chamber in order to investigate the effects of GSN itself on PMN chemotaxis. Cell counts in lower chamber were recorded in real time.Results:There was no obvious difference in neutrophils chemotaxis, when recombinant GSN was added into the lower chamber(P>0.05).Conclusions:GSN itself has no effect on PMN chemotaxis.Part ⅡObjective:To investigate the effects of extracellular GSN on N-formylmethionyl-leucyl-phenylalanine (fMLP) induced rat PMN chemotaxis.Methods:PMN were isolated from rat venous blood. To investigate the effects of extracellular GSN on fMLP induced PMN chemotaxis, PMN suspension and recombinant GSN was added to the upper chamber of transwell culture system, and fMLP was added to the lower chamber. Cell counts in lower chamber were recorded in real time.Results:There was no obvious difference in neutrophils chemotaxis, when recombinant GSN was added into the upper chamber(P>0.05).Conclusion:Extracellular geloslin exerts no effects on fMLP-induced PMN chemotaxis.Part IIIObjective:To investigate the effects of intracellular GSN on fMLP induced rat PMN chemotaxis.Methods:PMN were isolated from rat venous blood. Recombinant GSN or anti-GSN antibodies were transfected into freshly prepared PMN with electroporation method, and the chemotaxis function was observed respectively in transwell culture system, in order to investigate the effects of intracellular GSN level on PMN chemotaxis. Cell counts in lower chamber were recorded in real time.Results:There was no obvious difference in fMLP-induced neutrophils chemotaxis, when recombinant GSN electrotransfected into cytoplasm(P> 0.05); However, the rate of chemotaxis and chemotaxis index of neutriphils electrotransfected with anti-gelsolin antibodies was decreased significantly (P<0.05).Conclusion:Electrotransfecting with anti-gelsolin antibodies intracellular inhibits fMLP-induced rat neutrophils chemotaxis.