槲皮素降低邻苯二甲酸酯类混合物暴露致大鼠睾丸组织的氧化损伤的机制  

Mechanisms mediating the inhibitory effects of quercetin against phthalates-induced testicular oxidative damage in rats

作　　者：刘丽兰 邓儒雅 周稳进 林敏 夏玲姿 高海涛 LIU Lilan;DENG Ruya;ZHOU Wenjin;LIN Min;XIA Lingzi;GAO Haitao(Department of Preventive Medicine,School of Public Health and Management,Wenzhou Medical University,Wenzhou 325035,China;Zhejiang Provincial Key Laboratory of Watershed Science and Health,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]温州医科大学公共卫生与管理学院预防医学系,浙江温州325035 [2]温州医科大学浙江省流域水环境与健康风险研究重点实验室,浙江温州325035

出　　处：《南方医科大学学报》2023年第4期577-584,共8页Journal of Southern Medical University

基　　金：国家自然科学基金(81903321);浙江省自然科学基金(LY22H260002);温州市科技局项目(Y2020098)。

摘　　要：目的研究槲皮素(Que)对3种邻苯二甲酸酯类混合物(MPEs)暴露致大鼠睾丸组织氧化损伤的抑制作用及机制。方法40只雄性SD大鼠随机分对照组,MPEs组,MPEs联合低、中、高剂量Que组(MPEs+L-Que组、MPEs+M-Que组、MPEs+H-Que组)。MPEs组大鼠灌胃900 mg/(kg·d)MPEs,MPEs联合低、中、高剂量Que组大鼠灌胃900 mg/(kg·d)MPEs及10、30、90 mg/(kg·d)Que;对照组大鼠灌胃等容量的赋形剂;连续灌胃30 d后,麻醉大鼠,进行称重、采血、处死、量肛殖距、解剖、取睾丸和附睾等组织。试剂盒检测血清睾酮、促黄体生成素(LH)、卵泡刺激素(FSH)及睾丸组织丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT);通过H&E染色法观察睾丸组织病理结构;Western blot及免疫荧光检测睾丸组织中Nrf2、Keap1和HO-1的表达水平。结果与对照组相比,MPEs组大鼠肛殖距、睾丸、睾丸系数、附睾及附睾系数均减小(P<0.05);血清睾酮、FSH和LH水平下降(P<0.01);睾丸组织曲细精管萎缩,精细胞发育停滞,睾丸间质细胞增生等;睾丸组织中Nrf2、MDA、SOD、CAT和HO-1均升高(P<0.01),而Keap1降低(P<0.01);与MPEs组相比,MPEs+H-Que组大鼠睾丸、睾丸系数、附睾、附睾系数,血清睾酮、LH和FSH均升高(P<0.05);睾丸组织病理结构明显改善;MPEs+M-Que组和MPEs+H-Que组睾丸组织中MDA、SOD、CAT、Nrf2和HO-1水平下降(P<0.05),而Keap1升高(P<0.05)。结论Que可抑制MPEs暴露所诱导的大鼠睾丸组织氧化损伤,其机制可能与Que直接清除自由基,降低了MPEs诱导的睾丸组织氧化应激,恢复Nrf2信号通路的调节有关。Objective To explore the mechanism underlying the inhibitory effect of quercetin against testicular oxidative damage induced by a mixture of 3 commonly used phthalates(MPEs)in rats.Methods Forty male Sprague-Dawley rats were randomly divided into control group,MPEs exposure group,and MPEs with low-,median-and high-dose quercetin treatment groups.For MPEs exposure,the rats were subjected to intragastric administration of MPEs at the daily dose of 900 mg/kg for 30 consecutive days;Quercetin treatments were administered in the same manner at the daily dose of 10,30,and 90 mg/kg.After the treatments,serum levels of testosterone,luteinizing hormone(LH),follicle stimulating hormone(FSH),and testicular malondialdeyhde(MDA),catalase(CAT)and superoxide dismutase(SOD)were detected,and testicular pathologies of the rats were observed with HE staining.The expressions of nuclear factor-E2-related factor 2(Nrf2),Kelch-like ECH2 associated protein 1(Keap1)and heme oxygenase 1(HO-1)in the testis were detected using immunofluorescence assay and Western blotting.Results Compared with the control group,the rats with MPEs exposure showed significant reductions of the anogenital distance,weight of the testis and epididymis,and the coefficients of the testis and epididymis with lowered serum testosterone,LH and FSH levels(P<0.05).Testicular histological examination revealed atrophy of the seminiferous tubules,spermatogenic arrest,and hyperplasia of the Leydig cells in MPEs-exposed rats.MPEs exposure also caused significant increments of testicular Nrf2,MDA,SOD,CAT and HO-1 expressions and lowered testicular Keap1 expression(P<0.05).Treatment with quercetin at the median and high doses significantly ameliorated the pathological changes induced by MPEs exposure(P<0.05).Conclusion Quercetin treatment inhibits MPEs-induced oxidative testicular damage in rats possibly by direct scavenging of free radicals to lower testicular oxidative stress and restore the regulation of the Nrf2 signaling pathway.

关 键 词：槲皮素 邻苯二甲酸酯 睾丸 氧化损伤 Nrf2信号通路 

分 类 号：R285.5[医药卫生—中药学]