The Effect And Mechanism Of BFGF Mediated By ERK1/2Signaling Pathway On The Proliferation And Differentiation Of Human Lens Epithelial Cells

Objective: According to the experiment, cultured HLECsunder bFGF stimulation and blocking agent PD98059with ERK signalpathway, respectively, to detect the expression of α-SMA(alpha smoothmuscle actin) mRNA and phosphorylation, whether through ERK pathwayand discussion on related mechanism. Method: The human lens epithelialcells primarily cultured in vitro,were resuscited, passaged and cultured.The cultured cells passaged3-5generations, good growth, coveringculture bottle70-80%, which meets the requirement of theexperiment..Accession to the bFGF and specific blocker PD98059function for a certain period of time. The number and activity of HLECsare detected by MTT; reverse transcriptase polymerase chain reactiondetects the expressionof α-SMA mRNA in HLECs; Western blot is usedto detect the level of ERK1/2protein phosphorylation. All statistics werecalculated in SPAA software. In order to verify whether the ERK signalingpathway is involved in the formation of PCO.Result:1.the number ofHLECs in bFGF group were increased, the cell activity was enhanced, nosignificant differences between the two groups (P>0.05). The role of along time group, cell number more than short-term group, the difference was statistically significant (P<0.05); The group Only plus in bFGF thenumber of cells were significantly more than just add blocking agentgroup and blank control group, the difference was statistically significant(P<0.05); Both bFGF and PD98059were added, in blocking agent longaction groups, the amount and activity of cells below the groups effectshort visiblely, the difference was statistically significant (P<0.05).2.Thedose of bFGF is unchanged, as the extension of time, the alpha-SMAmRNA expression increased significantly, the difference was statisticallysignificant (P<0.05); In the bFGF action time of6hours, the alpha-SMAmRNA value reached the highest level, there is significant differencebetween two groups (P<0.05); Plus blocking agent PD98059, α-SMAreduced expression of mRNA and with the antagonist action timeincreased, no significant differences between the two groups (P>0.05).3.Inthe same bFGF dose,with the prolongation of time,the expression ofp-ERK gradually increased, Proved the secretion of bFGFcould promotephosphorylation. in the blank group, the expression ofnon-phosphorylation is more than phosphorylation.0.5h, phosphorylatedbegan more than non-phosphorylation.1hours later, phosphorylationexpression reached a peak at6hours, the expression of phosphorylatedand non-phosphorylated visible difference is not obvious.Conclusion:1. bFGF promot value-added effect on HLECs visible cell division phase.On the premise of constant dose, We can find that thepresentation time correlation.2. Alpha-SMA in normal human lens epithelial cells did not express, inthe bFGF concentration and action time, can form the detection of α-SMA, and showed time and dose dependence.3. ERK signaling pathway may be mediated by bFGF which may promotethe value-added effect on HLECs presents the temporal correlation,ERK signal pathway specific inhibitor PD98059can reduce theproliferation and activity of HLECs cells, inhibit α-SMA.