The Study Of TCDD Induced Transgenerational Inheritance Effect On The MRNA/lncRNA Profiling

Objective 2,3,7,8-tetrachlorodibenzo-p-dioxin?TCDD?can induce a phenotypic,intergenerational genetic effect.In this study,the mRNA and lncRNA expression profiles of livers from F1-F3 male rats exposed to intrauterine exposure to TCDD during the sex-determining stage of gonadal sex were studied.The bioinformatics analysis was performed to explore mRNA and lncRNA in the F1-F3 generation.Change between generations and the potential mechanism of action.Methods A transgeneration genetic model was established in rats.F0 female Sprague-Dawley rats were given gavage daily for 8-14 days?GD 8-14?.The doses were 0 and 200 ng/kg bw TCDD.The livers of F1-F3 ad?lt male rats were taken from 3 males in each group for a total of 18 samples.The mR NA and lncRNA were detected using Arraystar Rat LncRNA microarray to determine the differential expression profiles of m RNA and lnc RNA.The q RT-PCR method was used to detect F1 generations.Slc34a2,Zfp37 and Agxt2,F2 generation Bcl6,Aft3 and Adh4,F3 generation Ccnd1,Cyp7a1,Fcgbp,Hsd17b2,Aldh1a7,Lrtm2 and Trib3 mRNA expression levels and F1 generation XR₃47266,XR₅93760,XR₃43608 and XR₅92612,F2 generation uc.465-,XR₅89107 and XR₅95742,F3 generation XR₃61202,XR₅90171,XR₃60701 and XR₅95552 lncRNA expression levels;GO and KEGG pathway analysis of differential genes to predict the biological function of differentially expressed genes;subclass analysis of differentially expressed lncRNA,antisense antisense?antisense?lncRNA and their sense m RNAs were analysed together to deduce the function of the relevant antisense lncRNA and the differential lincRNAs were analyzed in conjunction with nearby mR NAs?< 300 kb?to deduce lincRNA function;the F1-F3 generation 200 Ng/kg bw TCDD-treated mR NA and lnc RNA were subjected to intergenerational differentially expressed gene enrichment analysis and cluster test(Se The inter-generational trend analysis of mRNA and lncRNA in the 200-ng/kg bw TCDD exposure group of the F1-F3 generation was performed to estimate the overall change in gene expression across generations.Res?lts 1.Prenatal TCDD exposure had different effects on liver mRNA expression profiles in F1,F2 and F3 male rats.The difference was that the differentially up-reg?lated genes involved in cytotoxic P450,tumorigenesis,and glucose and lipid metabolism in the F1 generation.Pathways;For the F2 generation,the differentially up-reg?lated genes were involved in pathogenesis of ad?lt onset diabetes and chemical carcinogenesis;F3 generations of differentially up-reg?lated genes were mainly involved in the immune pathway.2.The combined analysis of antisense LncRNA and corresponding mRNAs in the liver of F3 generation TCDD and control males showed that XR₅89480 and XR₅96950 may reg?late LOC367485 and Nrg2 mR NA at transcriptional or posttranscriptional levels through a variety of mechanisms.The combination of differentially expressed lincRNAs and adjacent mR NAs?< 300 kb?in the liver of F1-F3 generation TCDD-treated and control male rats can be used to infer differential expression of lincRNA function in each generation.3.Compared with the F1 generation group,the mR NA levels of the F2 generation group were mainly involved in the cytochrome P450 and IL-17 pathways;compared with the F2 generation group,the m RNA levels of the F3 generation group were down-reg?lated.It is mainly involved in PPAR and ins?lin pathways;compared with the F1 generation,the differences in the F3 generation of down-reg?lated mRNAs are mainly involved in the metabolism of glycerol lipids and the pathogenesis of ad?lt onset diabetes.Compared with the F3 generation control group,the F3 generation group showed upreg?lation of gene expression involved in the immune process,but compared with the F1 generation group,the F2 generation group was involved in down-reg?lation of the IL-17 pathway gene expression.The res?lts of liver mRNA analysis of male rats exposed to TCDD F1-F3 generation showed that the No.7 map had the largest number of differentially expressed mRNAs and 580 differentially expressed mRNAs?P<0.05?;in addition,the spectra 13 and 12 The mRNA expression levels of the two clusters increased with the TCDD exposure groups of the F1,F2,and F3 generations.The spectrum 9 increased first and then decreased.The spectrum 6 decreased first and then increased.Trend analysis of liverlncRNA in male rats exposed to TCDD from F1-F3 generation showed that the No.7 map had the largest number of differentially expressed lncRNA and 413 differentially expressed lncRNA?P<0.05?;in addition,spectra 13 and 8 The expression of lncRNA in the two clusters increased gradually from the TCDD exposure group of the F1,F2,and F3 generations.The spectrum 9 and the spectrum 7 increased first and then decreased.Conclusions1.Compared with the control group at each generation,the effect of prenatal TCDD exposure on liver mRNA expression in male F1,F2 and F3 rats was not the same.2.The effects of prenatal TCDD exposure on liver lncRNA expression in F1,F2 and F3 male rats were not the same compared to each generation control group.Among them,antisense LncRNA differentially expressed in F3 generation may be involved in the reg?lation of the corresponding mR NAs;different differentially expressed lincRNAs in F1-F3 generation may have similar biological functions as adjacent mR NAs.3.The overall gene profile of liver mR NA and lncRNA in F1-F3 generation TCDD-treated rats showed a consistent change,indicating that the effect of prenatal TCDD exposure across generations has a trend of change.Genetic effects of cytochrome P450 metabolism activation induced by TCDD may be diluted in the F2 generation;as the generation increases,the risk of developing ins?lin resistance and diabetes due to TCDD exposure may decrease;and TCDD exposure-mediated immunity Functional abnormalities may reappear in the F3 generation.These non-linear changes in gene expression along with increasing generations suggest that the genetic effects of EDCs across generations are complex.In summary,organisms may have the ability to correct the transmission of adverse genetic effects caused by exposure of EDCs in the process of evolution,but it is also possible that non-linear changes in genetic trends occur as the generations increase,and the inheritance of adverse effects may reappear in offspring.These phenomena provide new directions and ideas for further exploration of intergenerational inheritance of EDCs.