基于代谢组学探讨益肾通络方修复苯并(a)芘染毒大鼠精子DNA异常甲基化的机制  

Study on the mechanism of Yishen Tongluo Recipe in repairing abnormal methylation of sperm DNA in rats exposed to Benzo(a)pyrene based on metabolomics

作　　者：常贵峰 张宸铭[1] 王祖龙[2] 孙自学[3] 张琦[2] 陈如兵 华众 李凝华 赵士忠 申惠元 万雯熙 CHANG Gui-feng;ZHANG Chen-ming;WANG Zu-long;SUN Zi-xue;ZHANG Qi;CHEN Ru-bing;HUA Zhong;LI Ning-hua;ZHAO Shi-zhong;SHEN Hui-yuan;WAN Wen-xi(Henan University of Chinese Medicine,Zhengzhou,Henan 450046,China;First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou,Henan 450003,China;Second School of Clinical Medicine,Henan University of Chinese Medicine,Zhengzhou,Henan 450008,China)

机构地区：[1]河南中医药大学,河南郑州450046 [2]河南中医药大学第一附属医院,河南郑州450003 [3]河南中医药大学第二临床医学院,河南郑州450008

出　　处：《时珍国医国药》2023年第7期1537-1542,共6页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金面上项目(82174377,81974573,81603632);河南省科技攻关计划(192102310159);河南省自然科学基金(222300420485);国家中医临床研究基地科研专项(2021JDZX2056,2021JDZX2068)。

摘　　要：目的通过代谢组学探讨益肾通络方修复苯并(a)芘致精子DNA异常甲基化的作用机制。方法采用随机数字法将30只SPF级雄性SD大鼠,随机分为对照组、BaP染毒组及治疗组,每组大鼠各10只。除对照组外其余两组均按0.1mg/(kg·d)浓度的BaP进行染毒,持续60d;自BaP染毒第31天起,治疗组每日灌胃益肾通络方水煎液,共30d。末次给药后对3组大鼠实施乙醚吸入麻醉,取左侧睾丸制备睾丸样本,最后通过代谢组学技术对大鼠睾丸组织中代谢物的进行趋势分析及京都基因与基因组百科全书(KEGG)分析。结果经趋势分析后发现有41个代谢物呈现良好的回调趋势。与染毒组比较,益肾通络方可上调其中的8个代谢物物(P<0.05),下调其中的33个代谢物物(P<0.05)。通过富集分析发现其主要代谢通路为胆碱代谢通路;甘油磷脂代谢通路;维生素的消化和吸收;硫胺素代谢以及维生素B6代谢共5条代谢通路。结论益肾通络方对BaP染毒诱导的精子DNA异常甲基化的治疗作用可能与改善睾丸中维生素B6以及胆碱和甘油磷脂的代谢活动有关。Objective To investigate the mechanism of Yishen Tongluo Recipe in repairing the abnormal methylation of sperm DNA induced by benzo(a)pyrene based on metabolomics.Methods Thirty SPF male SD rats were randomly divided into control group,BaP exposure and treatment group by random number method,10 rats in each group.Except for the control group,the other two groups were exposed to BaP at 0.1 mg/(kg·d)for 60 d;the treatment group was given the decoction of Yishen Tongluo Recipe every day from the 31st day of BaP exposure.30d.After the last administration,three groups of rats were anesthetized with ether inhalation,and the left testicle was taken to prepare testicle samples.Finally,metabonomics technology was used to analyze the trend of metabolites in rat testis and Kyoto Encyclopedia of Genes and Genomes(KEGG).Results After trend analysis,41 metabolites were found to show a good trend of retracting.Compared with the exposure group,Yishen Tongluo Recipe could up-regulate 8 metabolites(P<0.05),and down-regulate 8 metabolites among them(P<0.05).33 metabolites(P<0.05).Through enrichment analysis,it was found that the main metabolic pathways were choline metabolism pathway;glycerophospholipid metabolism pathway;vitamin digestion and absorption;thiamine metabolism and vitamin B6 metabolism,a total of 5 metabolic pathways.Conclusion The therapeutic effect of Yishen Tongluo Recipe on the abnormal methylation of sperm DNA induced by BaP exposure may be related to the improvement of the metabolic activities of vitamin B6,choline and glycerophospholipid in the testis.

关 键 词：益肾通络方 精子DNA异常甲基化 代谢组学 代谢通路 

分 类 号：R285.5[医药卫生—中药学]