Based On LncRNA To Explore The Effect Of Yishen Tongluo Recipe On DNA Damage Of Sperm In Rats Exposed To Benzo (a) Pyrene

Objective:Explore the main biological pathways and signal pathways involved in sperm DNA damage in rats exposed to benzopyrene(Ba P);explore the effects of Yishen Tongluo recipe on sperm DNA damage in rats exposed to Ba P through lncRNA regulation.Method:1.30 SD rats were randomly divided into 3 groups: solvent control group,Ba P poisoning group and Yishen Tongluo prescription group,10 rats in each group.The Ba P stock solution was diluted with corn oil to a working solution with a concentration of 0.1 mg/ml.Except for the solvent control group at 9:00 on the day,all the other groups were given 1ml/kg intragastric dose at a concentration of 10mg/(Kg · d)for 60 days.In the solvent control group,1ml/kg corn oil was given by gavage.The rats in the Yishen Tongluo prescription group were given 10ml/(Kg · d)of Yishen Tongluo prescription from the 31 st day after Ba P exposure,each time about 1.044 g /ml,lasts 30 d.2.24 hours after the end of the poisoning,the epididymis was anesthetized with 0.9% sodium pentobarbital,the epididymis was collected,the epididymal tail sperm suspension was prepared,and the sperm DFI of each group was detected by flow cytometry.3.Select 3 rats from each group to detect the differentially expressed lncRNA and m RNA in the epididymal tissue of each group of rats by high-throughput sequencing,and perform GO analysis and pathway analysis between groups for the differential m RNA.4.Lnc RNA and m RNA correlation analysis,including antisense analysis,cis target gene prediction,trans target gene prediction,target gene function enrichment analysis.Result:1.The sperm DNA damage model of Bap-infected rats was successfully established.Compared with the solvent control group,the sperm DNA fragmentation index of rats in the Bap-exposed group increased significantly,and the difference was statistically significant.The sperm of the Yishen Tongluo prescription group The DNA fragmentation rate was lower than that of the Bap-exposed group,and the difference was statistically significant.2.The results of high-throughput sequencing showed that there were 195 differentially expressed lncRNAs obtained in the solvent control group and the Yishen Tongluo prescription group,of which 146 lncRNA expression increased,49 lncRNA expression decreased,and a total of 290 differentially expressed m RNAs,of which 162 The expression of two m RNAs increased,and the expression of 128 m RNAs decreased;there were 72 differentially expressed lncRNAs obtained in the solvent control group and the Bap-exposed group,of which 36 had increased expression and 36 had decreased expression,and a total of 199 differentially expressed m RNAs,93 of which were expressed There were 398 differentially expressed lncRNAs in the Bap-exposed group and the Yishen Tongluo prescription group,of which 304 had increased expression,94 had decreased expression,and a total of 631 differentially expressed m RNAs,of which 340 had increased expression,291 expressions decreased.3.The results of the correlation analysis between lncRNA and m RNA show that the difference genes are mainlyThere must be cell types,organelle types,cell parts types,and organelle parts types;in terms of participating in biological processes,genes are significantly enriched in cell process types,single tissue processes and metabolic processes;in terms of molecular functions,there are mainly binding molecules Function category and catalytic activity function category.The main pathways through which Cis acts on target genes include cell adhesion molecules,MAPK signaling pathways,endocytosis,and the interaction between cytokines and cytokine receptors;the main pathways through which target genes act on trans include neuroactive ligand-receptor interactions.,Interaction of cytokines and cytokine receptors and olfactory conduction,etc.Conclusion:1.Yishen Tongluo Recipe can reduce the rate of rat sperm DNA fragmentation,and has a certain protective effect on rat sperm DNA damage.2.The main biological pathways involved in DNA damage of Bap-infected rat sperm are cell process categories,cell categories,cell parts categories,and binding molecule functional categories;the main pathways are cell adhesion molecules,MAPK signaling pathways,endocytosis,The interaction of cytokines and cytokine receptors,neuroactive ligand-receptor interactions,olfactory conduction,etc.,lncRNA may play a regulatory role on the DNA damage of Bap-infected rat sperm through the above biological pathways and signaling pathways.