Preliminary study on the expression of long non-coding RNA in testis of oligoasthenospermia treated with Wuzi Yanzong Pill
Objective To study the intervention of Wuzi Yanzong Pill on long non coding RNA(lncRNA)in the testis related to oligoasthenozoospermia,and to preliminarily explore the potential mechanism of drug regulation of lncRNA in its intervention of oligoasthenozoospermia.Methods Establishment of oligoasthenospermia mouse model by intraperitoneal injection of busulfan,after 14 days of intervention with Wuzi Yanzong Pill,the number and vitality of epididymal sperm in small animals were measured using an automatic sperm detector,and the fertility was evaluated by mating with female mice,RNA was extracted from testicular tissues of 6 cases in the normal,model and drug groups,and transcriptomic sequencing technology was used to analyze and screen differentially expressed lncRNA regulated by Wuzi Yanzong Pill.Real-time fluorescent quantitative polymerase chain reaction(Real time PCR)was used to verify the differentially expression of lncRNA data.Bioinformatics was used to correlate the obtained drug-regulated lncRNA with the previously reported drug-regulated mRNAs,establish a coexpression network for drug regulated lncRNA targeting mRNAs,and screen the core mRNAs targeted by lncRNA in the network,which was performed by KEGG analysis,construct a drug regulated lncRNA targeted signaling pathway network and analyze the signaling pathway of Wuzi Yanzong Pill in regulating lncRNA intervention in oligoasthenozoospermia.Results Wuzi Yanzong Pill significantly improved the quality of sperm in oligoasthenozoospermia,the fertility rate of mating with female mice with oligoasthenozo-ospermia,obviously regulated 297 testicular lncRNA,of which 215 were up-regulated and 81 were down-regulated.The method of Real time PCR was used to confirmed the reliability of lncRNA sequencing results.The association analysis of drug regulated lncRNA mRNAs identified the core lncRNA(102)and mRNAs(81)regulated by drugs,lncRNA targeting signaling pathway association analysis showed that among 102 lncRNA,80 lncRNA targeted two or more signaling pathways,and 40 of the signaling pathways corresponded to two or more lncRNA,these lncRNA target signaling pathways that involved the entire cell cycle process of testicular cells,including growth,differentiation,metabolism,apoptosis,and signal transduction,and are mainly enriched in pathways such as the Fanconi anemia pathway,cell cycle,spliceosomes,FoxO pathway,p53 pathway,and homologous recombination etc.Conclusion This study provides the expression profile of lncRNA in the testes of Wuzi Yanzong Pill for intervention in oligoasthenozoospermia,these lncRNA were involved in drug repair of testicular spermatogenic damage,suggesting that they may play an important role in drug efficacy and are potential intervention targets for Wuzi Yanzong Pill.
万方数据
维普
