Effects of dexamethasone and latrunculin A on expression of protein in human trabecular meshwork cell

Background Researches have demonstrated that dexamethasone (Dex) can induce the changes of the function and structure of trabecular meshwork cells,and latrunculin A (Lat A) can enhance the outflow of aqueous humour and therefore low the intraocular pressure.Objective The aim of the present study is to investigate the effects of Dex and Lat A on the expression of protein in human trabecular meshwork cells.Methods Human trabecular meshwork cells were primarily cultured in DMEM using expand culture method and the fifth generation of cells were used to this experiment.Dex and/or Lat A were added in medium as 10~(-6)mol/L Dex group(Dex treating for 24 hours),Dex+Lat A group(10~(-6)mol/L Dex+2mmol/L Lat A for 24 hours),Lat A group(2mmol/L Lat A for 24 hours) and DMEM culture group.Two dimensional gel electrophoresis(2 DE) was used to compare the protein expressions among these four groups.Subsequently protein spots with different intensity were selected for mass spectrometry analysis.Results Four gel patterns of two dimensional gel electrophoresis of human trabecular meshwork cells from Dex,Dex+Lat A,Lat A and control groups were obtained.A good isolated result for majority of proteins in human trabecular meshwork cells was found in all of the four groups.An obvious expression difference of proteins in human trabecular meshwork cells was seen among the different culture conditions.Twenty four kinds proteins were identified by GDPiMALDI TOF MS,including cytoskeleton related proteins,heat shock proteins,redox related proteins,and proteins participating in carbohydrate metabolism.The expressions of aldehyde dehydrogenase(ADLH)and Rab were increased in Lat A group and decreased in Dex group,but HSP27 and hCRMP2 showed the contrary outcome.Conclusion This study construct the pattern of protein expression in human trabecular meshwork cells by using 2 DE.Dex and Lat A impact the protein expressions in human trabecular meshwork cells. Key words: trabecular meshwork cells; proteomics; dexamethasone; latrunculin A