[Protective effect of Yishen Tongluo Recipe against benzo(a)pyrene-induced sperm DNA methylation changes in male rats].

ABSTRACT

OBJECTIVE: To explore the protective effect of Yishen Tongluo Recipe (YTR) against aberrant sperm DNA methylation in male rats exposed to benzo(a)pyrene (BaP). METHODS: Thirty male SD rats of the SPF grade were randomly divided into three groups of equal number solvent control, BaP exposure and YTR intervention. The animals of the solvent control group were injected intraperitoneally with 0.5% DMSO while those of the other two groups with BaP at 0.1 mg/kg/d, all for 60 days, and at 31 days of BaP exposure, those of the YTR group were treated intragastrically with YTR for 30 days. Then, the left epididymides were harvested from all the rats and sperm suspensions collected and centrifuged for extraction of sperm DNA. The methylated DNA immunoprecipitation sequencing (MeDIP-seq) technique was used to detect the whole-genome DNA methylation in different groups. RESULTS: Exposure to BaP induced the up-regulation of 828 genes encoding mRNA in the sperm DNA, while YTR intervention produced a significant protective effect on the transforming growth factor ß3 (TGF-ß3), cystic fibrosis transmembrane conductance regulator (CFTR) and recombination activating gene 1 (RAG1), and down-regulated the expressions of 3 227 genes. BaP exposure also caused the up-regulation of 783 genes encoding lncRNA in the sperm DNA, and YTR treatment exhibited an evident protective effect on 62 of the up-regulated genes, induced the down-regulation of 3 378 genes, and showed a protective effect on 56 of the down-regulated genes. CONCLUSIONS: YTR has a protective effect against aberrant sperm DNA methylation in male rats exposed to BaP, which may be associated with lncRNA.