丙硫氧嘧啶通过内质网应激损害大鼠生精功能和精子质量的机制及疏肝补肾毓麟汤的干预作用  

Mechanisms of Spermatogenic Dysfunction and Sperm Quality Degradation Induced by Propylthiouracil via Endoplasmic Reticulum Stress in Rats and Interventional Effect of Shugan Bushen Yulin Decoction

作　　者：孟宇豪 江晓翠 萧闵 王朝阳[1,2] MENG Yuhao;JIANG Xiaocui;XIAO Min;WANG Chaoyang(School of Basic Medical Sciences,Hubei University of Chinese Medicine,Wuhan 430065,China;Hubei Shizhen Laboratory,Wuhan 430065,China;Laboratory Animal Center,Hubei University of Chinese Medicine,Wuhan 430065,China)

机构地区：[1]湖北中医药大学基础医学院,武汉430065 [2]湖北时珍实验室,武汉430065 [3]湖北中医药大学实验动物中心,武汉430065

出　　处：《中国实验方剂学杂志》2025年第9期79-89,共11页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：湖北省自然科学基金中医药创新发展联合基金项目(2024AFD223);湖北省自然科学基金面上项目(2023AFB990);湖北省中医药管理局2023—2024年度中医药指导性项目(ZY2023Z204)。

摘　　要：目的:基于内质网应激(ERS)通路探讨疏肝补肾毓麟汤改善丙硫氧嘧啶(PTU)导致大鼠睾丸生精功能障碍及精子质量下降的作用机制。方法:使用随机分组法将60只雄性大鼠分为空白组,模型组,疏肝补肾毓麟汤低、中、高剂量组及左卡尼汀组,每组10只。除空白组外,其余5组连续12 d灌胃PTU(10 mg·kg^(-1))进行造模。造模完成后,疏肝补肾毓麟汤低、中、高剂量组(6.75、13.5、27 g·kg^(-1))分别给予疏肝补肾毓麟汤灌胃给药,左卡尼汀组给予左卡尼汀(0.27 g·kg^(-1))灌胃给药,空白组灌胃生理盐水,连续28 d。末次给药治疗24 h后取材,称体质量及睾丸、附睾、精囊腺质量,计算三者脏器指数;苏木素-伊红(HE)染色观察睾丸和附睾的病理变化,并对大鼠的睾丸生精功能进行评分;酶联免疫吸附测定法(ELISA)检测大鼠血清促甲状腺激素(TSH)、三碘甲状腺原氨酸(T3)、四碘甲状腺原氨酸(T4)、促卵泡素(FSH)、促黄体生成素(LH)、睾酮(T)、雌二醇(E_(2))、丙二醛(MDA)水平及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性;原位末端标记法(TUNEL)检测睾丸细胞凋亡率;蛋白免疫印迹法(Western blot)检测大鼠睾丸组织葡萄糖调节蛋白78(GRP78)、C/EBP同源蛋白(CHOP)、磷酸化蛋白激酶R样内质网激酶(p-PERK)、蛋白激酶R样内质网激酶(PERK)、磷酸化真核翻译起始因子2α激酶(p-EIF2α)、真核翻译起始因子2α激酶(EIF2α)、激活转录因子4(ATF4)蛋白表达水平。结果:与空白组比较,模型组大鼠睾丸、附睾、精囊腺体积减小,且3个器官的脏器指数显著降低(P<0.01);大鼠睾丸与附睾病理形态损害,生精功能显著下降(P<0.01);精子密度及精子活动率显著降低(P<0.01);大鼠血清TSH、MDA水平升高,T3、T4、FSH、LH、T、E_(2)水平及SOD、GSH-Px活性明显降低(P<0.05,P<0.01);大鼠睾丸细胞凋亡水平显著增加(P<0.01);大鼠睾丸组织GRP78、CHOP、p-PERK/PERK、p-EIF2�Objective:To investigate the mechanisms through which Shugan Bushen Yulin decoction ameliorates spermatogenic dysfunction and sperm quality degradation caused by propylthiouracil(PTU)via the endoplasmic reticulum stress pathway in rats.Methods:Sixty male rats were randomly assigned into six groups:a control group,a model group,low-(6.75 g·kg^(-1)),medium-(13.5 g·kg^(-1)),and high-dose(27 g·kg^(-1))Shugan Bushen Yulin decoction groups,and an L-carnitine(0.27 g·kg^(-1))group,with ten rats in each group.Other groups except the control group were administrated with PTU(10 mg·kg^(-1))by gavage for 12 consecutive days.After the completion of modeling,rats were administrated with corresponding agents or normal saline(control group)via gavage for 28 consecutive days.Twenty-four hours after the last administration,rats were sacrificed,and the body and organ(testis,epididymis,and seminal vesicle)weights were measured to calculate the organ indices.Hematoxylineosin staining was employed to observe the pathological changes in the testes and epididymis,and the testicular spermatogenic function of rats was scored.Enzyme-linked immunosorbent assay was employed to measure the levels of thyroid-stimulating hormone(TSH),triiodothyronine(T3),thyroxine(T4),follicle-stimulating hormone(FSH),luteinizing hormone(LH),testosterone(T),estradiol(E_(2)),and malondialdehyde(MDA)and the activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in the rat serum.Terminal deoxynucleotidyl transferase dUTP nick end labeling was adopted to assess the rate of testicular cell apoptosis.Western blot was conducted to determine the expression levels of glucose-regulated protein 78(GRP78),CCAAT/enhancer-binding protein homologous protein(CHOP),phosphorylated protein kinase RNA-like endoplasmic reticulum kinase(p-PERK),protein kinase RNA-like endoplasmic reticulum kinase(PERK),phosphorylated eukaryotic translation initiation factor 2αkinase(p-EIF2α),eukaryotic translation initiation factor 2αkinase(EIF2α),and activating transcript

关 键 词：疏肝补肾毓麟汤 丙硫氧嘧啶 内质网应激 蛋白激酶R样内质网激酶(PERK)/真核翻译起始因子2α激酶(EIF2α)/激活转录因子4(ATF4)信号通路 精子质量 

分 类 号：R242[医药卫生—中医临床基础] R289[医药卫生—中医学] R278