Infrared Spectroscopy Was Used To Analyze The Reproductive Toxicity Effects Of Nonylphenol And Bisphenol A

ObjectiveIn this study,nonylphenol(NP)and bisphenol A(BPA)were used as test substances,and SD male rats and Leydig cells(LCs)were used as research objects,and Fourier transform infrared spectroscopy coupled with multivariate statistical analysis methods(PCA-LDA)were applied to explored the male reproductive toxicity effects and joint toxicity of NP and BPA.Methods1.Four groups were set: control group(corn oil),NP exposure group(5 mg/kg NP),BPA exposure group(0.5 mg/kg BPA),and combined exposure group(5 mg/kg NP & 0.5 mg)/kg BPA),21-day-old father and mother SD rats were exposed every other day.When the father rats were 90 days old,the testicular tissue was taken,and each tissue was cut in half,half of which was prepared for hematoxylin and eosin staining,and the other half was prepared for FTIR testing.This batch was called F0P90+ group.2.When the male and female Sprague-Dawley rats grow to about 55 days of age,they were mated.The mother rats were exposed every other day during pregnancy and lactation.After weaning period,the progeny male rats are divided into two batches,one batch continued to be exposed to the next day,was called for the F1P90+ group;the other batch stopped the exposure,was called for the F1P90-group.when the male rat of the first generation grew to 90 days of age,the testicular tissue was taken,and each tissue was cut in half,half of which was prepared for hematoxylin and eosin staining,and the other half was prepared for FTIR testing.3.Three groups of Leydig tissue(F0P90+ group,F1P90+ group and F1P90-group)were subjected to FTIR spectroscopy;4.Principal component analysis and linear discriminant analysis(PCA-LDA)were used to analyze the data in the MATLAB R2017 a software.Result1.The organ coefficients of the testis and epididymis of each rats were calculated statistically and compared.It was found that the testis coefficient of the NP-treated group in F1P90+ group was significantly different from that of the control group(P<0.01).2.The characteristics of tissues in hematoxylin and eosin‐stained samples showed that the morphology of the testicular Leydig tissue in the exposed group changed,and the tightness of the connection with the seminiferous tubules was also reduced.3.LD1 and LD2 space were used to analyze the degree of separation between exposed groups and the control group.In fingerprint region,all the exposed group were significantly different from that of the control group(P<0.01),except for the F0P90+ NP exposure group.In the lipid region,except for the F1P90+ NP-treated group,the BPA-treated group and the F1P90-NP-treated group,the other groups were significantly different from the control group(P<0.001).Clustering vector analysis was used to analyze the main factors causing the difference between the exposed group and the control group.It was found that the main factors affecting the difference between the exposed group and the control group were protein phosphorylation,protein secondary structure,fatty acids and lipids.4.Under NP and BPA exposure,the lipid-to-protein ratio,lipid oxidation level,amide I to amide II ratio and acetylation level of Leydig cells in the three groups of rats all occurred changes.Conclusion1.The toxic effect of NP and BPA on testicular mesenchymal cells is mainly due to the destruction of cell membrane structure and integrity,altering the permeability and permeability of the cell membrane,leading to easier entry of NP and BPA into cells,enhancing toxic effects and presenting the joint effect.2.Long-term NP and BPA interventions inhibit the proliferation and differentiation of Leydig cells;NP and BPA can disrupt the process of lipid oxidative degradation of Leydig cells;after NP and BPA exposure ceases,cells can self-repair The normal level of protein consumption is restored,and the level of acetylation affected by NP and BPA is not self-recovery by the cells.