基于ERK/Nrf2-ARE通路探究枸杞多糖对七氟烷麻醉诱导雄性大鼠生殖毒性的拮抗作用机制  

Study on the mechanism of Lycium barbarum polysaccharide on reproductive toxicity induced by sevoflurane anesthesia in male rats based on ERK/Nrf2-ARE pathway

作　　者：孙颖 刘庄[2] 王雪楠[1] Sun Ying;Liu Zhuang;Wang Xuenan(Department of Anesthesiology,Affiliated Hospital of Jining Medical University,Jining 272002,China)

机构地区：[1]济宁医学院附属医院麻醉科,济宁272002 [2]济宁医学院附属医院生殖医学科,济宁272002

出　　处：《中华生殖与避孕杂志》2023年第8期806-812,共7页Chinese Journal of Reproduction and Contraception

基　　金：山东省中医药科技发展计划项目(2019-0463)。

摘　　要：目的探究枸杞多糖(Lycium barbarum polysaccharides,LBP)对七氟烷麻醉诱导的雄性大鼠生殖毒性的作用,分析其对细胞外信号调节激酶(extracellular-signal-regulated kinase,ERK)/核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)-抗氧化反应元件(antioxidant response element,ARE)信号通路的影响。方法取SD雄性大鼠按计算机随机生成数字法分为空白对照组、模型组、LBP低剂量组、LBP高剂量组和阳性对照组,除空白对照组外其余均以七氟烷麻醉诱导,同时腹腔注射给药,空白对照组和模型组予以等体积生理盐水,LBP低、高剂量组分别予以100 mg/kg、200 mg/kg LBP,每天1次,阳性对照组予以10 mg/kg维生素E,隔天1次,14 d后处死。计算各组大鼠睾丸与附睾的脏器指数;检测睾丸组织标志酶酸性磷酸酶(acid phosphatase,ACP)、碱性磷酸酶(alkaline phosphatase,AKP)、乳酸脱氢酶(lactate dehydrogenase,LDH)含量;HE染色观察大鼠睾丸组织病理学;检测睾丸组织中超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)含量;TUNEL染色观察大鼠睾丸组织细胞凋亡率;Western blotting检测大鼠睾丸组织内Bax、Bcl-2及ERK/Nrf2-ARE通路蛋白表达。结果LBP低、高剂量组和阳性对照组睾丸脏器指数[(0.39±0.04)%、(0.47±0.05)%、(0.62±0.05)%]和附睾脏器指数[(0.08±0.01)%、(0.11±0.01)%、(0.14±0.02)%]均比模型组[(0.33±0.03)%,(0.05±0.01)%]高(均P<0.001),ACP[(122.13±5.39)U/g、(115.37±4.45)U/g、(109.74±4.73)U/g]、LDH含量[(260.43±15.18)U/g、(245.17±8.13)U/g、(236.19±10.23)U/g]均比模型组[(129.98±6.17)U/g、(279.89±18.41)U/g]低(均P<0.001),AKP含量[(224.41±11.69)U/g、(247.59±12.17)U/g、(266.74±13.78)U/g]、SOD活性[(3.11±0.12)U·mg/蛋白、(4.05±0.14)U·mg/蛋白、(4.78±0.15)U·mg/蛋白]均比模型组[(200.48±10.48)U/g、(2.02±0.09)U·mg/蛋白]高(均P<0.001),MDA含量[(14.86±1.41)nmol·mg/蛋白、(10.39±1.22)nmol·mg/蛋白、(5.25±0.47)nmolObjective To explore the effect of Lycium barbarum polysaccharide(LBP)on sevoflurane anesthesia-induced reproductive toxicity in male rats,and to analyze its effect on extracellular signal-regulated kinase(ERK)/nuclear factor E2-related factor 2(Nrf2)-antioxidant response element(ARE)signaling pathway.Methods SD male rats were randomly divided into blank control group,model group,low-dose LBP group,high-dose LBP group and positive control group.Except for the blank control group,the rest were induced by sevoflurane anesthesia and administered by intraperitoneal injection at the same time.The model group was given equal volume of normal saline;the LBP low-and high-dose groups were given 100 mg/kg and 200 mg/kg LBP,once a day,respectively.The positive control group was given 10 mg/kg vitamin E,once every other day.After 14 d of the experiment,all rats were sacrificed.The organ indexes of testis and epididymis of rats in each group were calculated.The contents of testis tissue marker enzymes acid phosphatase(ACP),alkaline phosphatase(AKP)and lactate dehydrogenase(LDH)were detected.HE staining was used to observe the testicular tissue disease of rats.The contents of superoxide dismutase(SOD)and malondialdehyde(MDA)in testis tissue were detected.TUNEL staining was used to observe the apoptosis rate of rat testis tissue.Western blotting was used to detect Bax,Bcl-2 and ERK/Nrf2-ARE pathway protein expression level.Results In the low-dose and high-dose LBP groups and positive control group,the testicles[(0.39±0.04)%,(0.47±0.05)%,(0.62±0.05)%]and epididymal organ index[(0.08±0.01)%,(0.11±0.01)%,(0.14±0.02)%]were higher than those of the model group[(0.33±0.03)%,(0.05±0.01)%,all P<0.001],ACP[(122.13±5.39)U/g,(115.37±4.45)U/g,(109.74±4.73)U/g]and LDH content[(260.43±15.18)U/g,(245.17±8.13)U/g,(236.19±10.23)U/g]were lower than those in the model group[(129.98±6.17)U/g,(279.89±18.41)U/g,all P<0.001],the AKP content[(224.41±11.69)U/g,(247.59±12.17)U/g,(266.74±13.78)U/g]and SOD activity[(3.11±0.12)U·mg/pro

关 键 词：枸杞多糖 七氟烷 麻醉诱导 生殖毒性 睾丸 凋亡 

分 类 号：R285.5[医药卫生—中药学]