The Effects Of Lead Exposure On The Expression Of IGF1R,IGFBP3,A?40 And A?42 In PC12 Cells

Objective:To investigate the effects of lead exposure and IGF1 R inhibitor AG1024 on the expression of IGF1 and IGFBP3 in PC12 cells.It is clear that the mechanism of the related proteins inducing AD is regulated by them,thus providing theoretical guidance for the prevention and treatment of lead poisoning.Method:Use PC12 neuron cell line to cultivate and establish a corresponding lead exposure model,deal with cells with different concentrations of lead acetate respectively,divide the experiment into control group,1?mol/L Pb Ac,10?mol/L Pb Ac group,IGF1 R inhibitor(AG1024)group,IGF1 R inhibitor group(AG1024)+ 1?mol/L Pb Ac group,IGF1 Rinhibitor group(AG1024)+10?mol/L Pb Ac group,respective contamination's three periods of time 24h?48h and 72 h,use MTT method to determine the effect of lead exposure dose on cell proliferation,through western blot and immunocytochemistroy,determine the protein expression o f IGF1 R and IGFBP3 in PC 12 cells,use ELISA method to determine the expression of A?40 and A?42 in cell supernatant.Result: MTT result shows that during three time periods,using different concentrations of Pb Ac to deal with cells shows a negative correlation between OD490 values and exposure concentrations in the treatment group,the exposure concentration becomes higher,the absorbance values decrease significantly,compared with the control group,in the same treatment time groups,only the low-dose exposure 0.1 ?mol/L group has no significant difference,and the other differences have statistic significance(P<0.05);Western blot result shows that compared with the control group,there is no significant difference in IGF1 R protein expression in the low-dose lead exposure group(0.1 ?mol/L),(P>0.05),the expression of IGF1 R protein in the high-dose lead acetate group is significantly lower than that in the control group,the difference has statistic significance,P<0.05,the exposure of medium and high concentrations of lead significantly inhibits the expression of IGF1 R protein in the cell system,the greater the increase in exposure dose,the more obvious the inhibitory effect.Immunohistochemical staining result shows that the immunofluorescence positive reaction of IGF1 R cells is mainly positioned in the cytoplasm,and the average optical density and neuronal activity of IGF1 R immunofluorescent antibody at 10 ?mol/L dose in lead exposure group is significantly lower than that in the control group,the difference has statistical significance(P<0.05),and the downtrend of significant difference is more obvious;AG1024 inhibitor and lead combination(1 ?mol/L)and(10 ?mol/L) decrease significantly compared with the control group,the inter-group has statistical significance(P<0.05);the IGFBP3 protein activity in the lead exposure group is significantly lower than that of the IGFBP3 protein in the control group.the average optical density and neuronal activity of IGF1 R immunofluorescent antibody at 10?mol/L dose in lead exposure group is significantly lower than that in the control group,the difference has statistical significance(P<0.05).ELISA result shows that compared the lead exposure group with the control group,the content of A?40 and A?42 in cell supernatant has no statistical significance among the groups,(P>0.05).Conclusion:Lead can obviously down-regulate the expression of IGF1 R and IGFBP3,lead and inhibitor can inhibit the proliferation of cells,promote the tendency of apoptosis and damage the nervous system.