非瑟酮调节LKB1-AMPK-mTOR-p70S6K通路改善大鼠少弱精子症  

Mechanism analysis of fisetin regulating LKB1-AMPK-mTOR-p70S6K pathway to improve oligonasthenospermia in rats

作　　者：陈利帮 沈炳香 何春远 赵为陈 常伟 王桐生[4] CHEN Li-bang;SHEN Bing-xiang;HE Chun-yuan;ZHAO Wei-chen;CHANG Wei;WANG Tong-sheng(Dept of Clinical Sciences,Wanxi College of Health Professions,Lu’an Anhui 237000,China;Dept of Pharmacy,Lu'an Hospital Affiliated to Anhui Medical University,Lu’an Anhui 237005,China;School of Public Health,Anhui Medical University,Hefei 230032,China;Dept of Physiology and Pharmacology,School of Integrative Medicine,Anhui University of Traditional Chinese Medicine,Hefei 230013,China)

机构地区：[1]皖西卫生职业学院临床系,安徽六安237000 [2]安徽医科大学附属六安医院药学部,安徽六安237005 [3]安徽医科大学公共卫生学院,安徽合肥230032 [4]安徽中医药大学中西医结合学院生理与药理学系,安徽合肥230013

出　　处：《中国药理学通报》2024年第7期1296-1301,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81473674);安徽省高等学校科学研究项目(No 2022AH053045)。

摘　　要：目的研究非瑟酮对少弱精子大鼠的睾丸及精子保护作用,并探究其机制。方法构建大鼠少弱精子模型,将大鼠随机分为空白组、模型组、非瑟酮低、中、高剂量治疗组、LKB1激动剂组,每组各10只。ELISA法检测各组卵泡刺激素(follicle-stimulating hormone,FSH)、黄体生成素(luteinising hormone,LH)、睾酮(testosterone,T)、雌二醇(estradiol,E2)、催乳素(prolactin,PRL)水平;流式细胞术检测精子细胞凋亡;HE染色检测大鼠睾丸组织损伤;透射电镜检测精子细胞超微结构;qRT-PCR和Western blot检测LKB1、AMPK、mTOR、p70S6K mRNA和蛋白表达。结果与空白组相比,模型组和LKB1激动剂组FSH、LH、PRL、T等激素水平明显降低,精子细胞出现凋亡,睾丸损伤严重,LKB1、p-AMPK/AMPK表达明显上调,mTOR、p-p70S6K/p70S6K表达下调(P<0.05);与模型组相比,不同剂量的非瑟酮治疗后,精子细胞凋亡数明显减少,FSH、LH、PRL、T等激素水平明显上升,LKB1、p-AMPK/AMPK明显下调,mTOR、p-p70S6K/p70S6K明显上调(P<0.05)。结论非瑟酮可有效治疗少弱精症大鼠,其作用机制可能与LKB1-AMPK-mTOR-p70S6K信号通路有关。Aim To investigate the protective effect of fisetin on testis and sperm of rats with oligoasthenospermia and to explore its mechanism.Methods The rat model of oligoasthenospermia was established.The rats were randomly divided into the blank group,model group,low-,medium-,and high-dose fisetin treatment groups,and LKB1 agonist group,with 10 rats in each group.ELISA was used to detect the levels of FSH,LH,T,E2 and PRL.Flow cytometry was used to detect sperm cell apoptosis.HE staining was used to detect testicular tissue damage.Transmission electron microscopy was used to detect the ultrastructure of sperm cells.qRT-PCR and Western blot were used to detect the mRNA and protein expression of LKB1,AMPK,mTOR,and p70S6K.Results Compared with the blank group,the levels of FSH,LH,PRL,T and other hormones in the model group and LKB1 agonist group were significantly reduced,and sperm cell apoptosis and testicular injury were severe.The expressions of LKB1 and p-AMPK/AMPK were significantly up-regulated,while the expressions of mTOR and p-p70S6K/p70S6K were significantly down-regulated(P<0.05).Compared with the model group,after different doses of fisetin treatment,the number of apoptotic sperm cells was significantly reduced,the levels of FSH,LH,PRL,T and other hormones markedly increased,the expression of LKB1 and p-AMPK/AMPK was significantly down-regulated,and the expression of mTOR and p-p70S6K/p70S6K was evidently up-regulated(P<0.05).Conclusion Fisetin is effective in the treatment of oligoasthenospermia rats,which may be related to LKB1-AMPK-mTOR-p70S6K signaling pathway.

关 键 词：非瑟酮 少弱精症 AMPK MTOR P70S6K LKB1 

分 类 号：R-332[医药卫生] R321.1R322.64R329.25R698.2