还原型谷胱甘肽对氟中毒小鼠附睾的影响

试验旨在研究还原型谷胱甘肽(reduced glutathione,GSH)对氟中毒小鼠附睾的影响。将50只3周龄ICR雄性小鼠随机分为5组,分别为对照组和4组染氟组。对照组饮用蒸馏水,染氟组分别供给含100mg/L NaF的蒸馏水,自由饮水30d。之后在染氟组中选择3组每日分别灌胃200、400和800mg/(kg·只)GSH,同时各组小鼠再自由饮含氟水30d。试验结束后,摘取小鼠附睾进行活性氧(ROS)、丙二醛(MDA)、GSH含量及谷胱甘肽过氧化物酶(GPx)、谷胱甘肽硫转移酶(GSTs)和谷胱甘肽还原酶(GR)活性检测,同时制作石蜡切片,HE染色后进行组织形态结构分析。结果显示,与对照组相比,100mg/L NaF组精子密度、精子活力、GSH含量、GPx和GR活性均显著下降(P0.05),而GSH添加组较100 mg/L NaF组均有所升高;100 mg/L NaF组精子畸形率、ROS和MDA含量较对照组均显著升高(P0.05),而GSH添加组较100mg/L NaF组均降低,其中800mg/kg GSH组效果明显。与对照组相比,100mg/L NaF组附睾头、附睾体及附睾尾管腔厚度升高,管腔内精子数量下降;GSH添加组附睾头、附睾体及附睾尾的管腔厚度与管腔内精子数量都有所恢复,其中800mg/kg GSH组效果较好。综上所述,添加GSH后氟中毒小鼠谷胱甘肽抗氧化系统酶活性有所升高,附睾组织形态结构一定程度上有所恢复,对附睾有一定的改善作用。     

还原型谷胱甘肽对氟中毒小鼠睾丸的影响

研究不同剂量还原型谷胱甘肽(GSH)对氟中毒小鼠睾丸的影响。将50只3周龄ICR雄性小鼠随机分为5组,其中1组为对照组,其余4组为高氟组。对照组给予蒸馏水,染氟组给含100mg/L NaF的蒸馏水,自由饮用30d。在染氟组中选取3组分成100mg/L NaF+200mg/kg GSH组、100mg/L NaF+400mg/kg GSH组和100mg/L NaF+800mg/kg GSH组,这3组小鼠每只每日分别以200、400、800mg/kg GSH的剂量进行灌胃,同时这5组小鼠再自由饮含氟水30d。饲养结束后,摘取小鼠睾丸进行活性氧(ROS)、丙二醛(MDA)、GSH含量及谷胱甘肽过氧化氢酶(GPx)、谷胱甘肽硫转移酶(GSTs)、谷胱甘肽还原酶(GR)的活性检测;制作石蜡切片,HE染色进行组织形态结构分析。结果显示,与对照组相比,100mg/L NaF组ROS和MDA含量显著升高,GSTs和GR活性显著降低;与100mg/L NaF组相比,添加GSH组100mg/L NaF+800mg/kg GSH组GR活性显著升高。与对照组相比,100mg/L NaF组曲细精管结构稀疏,细胞壁细胞层厚度变小,各级生精细胞排列紊乱,还有部分生精细胞脱落,管腔半径变大且管腔中央精子数量明显减少;随着GSH浓度的增加,100mg/L NaF+800mg/kg GSH组得到最大程度的恢复。添加GSH后氟中毒小鼠谷胱甘肽抗氧化系统酶活性有所升高,睾丸组织形态结构一定程度上有所恢复,对睾丸功能有一定的改善作用。     

氟中毒对小鼠肾脏ROS、MDA和GSH含量的影响

为了研究氟中毒对小鼠肾脏活性氧(ROS)、丙二醛(MDA)和还原型谷胱甘肽(GSH)含量的影响,并研究添加GSH后是否有缓解作用,试验建立氟化钠(NaF)+GSH模型,将50只小鼠分为对照组、100 mg/L NaF组、100 mg/L NaF+200 mg/kg GSH组、100 mg/L NaF+400 mg/kg GSH组和100 mg/L Na F+800 mg/kg GSH组,饲养结束后断颈处死小鼠,采集肾脏测定ROS、MDA和GSH含量。结果表明:与对照组比较,100 mg/L Na F可极显著增加肾脏MDA含量,降低GSH含量(P 0. 01);加入GSH后,与100 mg/L NaF组比较,100 mg/L NaF+800 mg/kg GSH组的GSH含量显著增多(P0. 05);而对于ROS,氟摄入和添加GSH均未发现明显变化(P0. 05)。说明添加GSH可以缓解氟中毒引起的肾脏损伤。     

氟对小鼠睾丸谷胱甘肽抗氧化系统的影响

为探讨氟化钠对小鼠睾丸谷胱甘肽抗氧化系统的影响,将24只3周龄ICR雄性小鼠随机分为对照组和染氟组,对照组给予蒸馏水,染氟组分别供给含25、50、100mg·L~(-1) NaF的蒸馏水,自由饮用60d。染毒结束后检测小鼠睾丸总抗氧化能力(T-AOC)、活性氧(ROS)、丙二醛(MDA)、还原型谷胱甘肽(GSH)的含量及谷胱甘肽过氧化氢酶(GPx)、谷胱甘肽硫转移酶(GST)、谷胱甘肽还原酶(GR)的活性,检测GPx1、GPx4、GST-mu5和GR基因的转录。电镜观察睾丸组织细胞内线粒体、内质网和高尔基体的结构。结果显示,氟中毒可使睾丸T-AOC减弱,ROS、MDA含量升高,GSH含量显著降低,细胞内氧化水平升高,线粒体、内质网和高尔基体出现不同程度的损伤,GPx、GST、GR的活性降低,GPx1、GPx4、GST-mu5和GR基因转录下调。小鼠氟中毒后,睾丸抗氧化能力降低,处于明显的氧化应激状态,这与谷胱甘肽抗氧化系统酶活性降低及其相关基因转录下调有关。     

氟化钠对附睾形态结构及紧密连接相关基因表达的影响

为了研究氟对小鼠附睾结构形态及紧密连接相关基因的影响,将60只健康雄性小鼠随机分成4组,对照组饮用蒸馏水,3个氟处理组分别饮用含氟化钠（NaF） 25、50和100 mg/L的蒸馏水。染氟60 d,取小鼠股骨,采用氟离子选择电极法测其氟含量;取右侧附睾制作石蜡切片,HE染色后进行组织形态结构观察;左侧附睾提取RNA,采用实时荧光定量PCR技术（qRT-PCR）检测紧密连接相关基因的表达。结果显示,与对照组相比,氟处理组小鼠股骨氟含量随摄氟剂量的增加呈剂量依赖性,其中50、100 mg/L剂量组差异极显著（P<0.01）;氟处理组附睾尾管腔厚度极显著增厚（P<0.01）;紧密连接相关基因Claudin-2 mRNA表达量极显著下降（P<0.01）,100 mg/L剂量组ZO-1 mRNA表达量显著下降（P<0.05）。摄入过量NaF影响附睾形态及其紧密连接相关基因的表达,从而可能对血-附睾屏障造成损伤。     

母鼠围产期氟暴露对仔鼠肾脏病理变化的研究

本文以昆明小鼠建立动物模型,按照雌∶雄=2∶1将小鼠随机分为四组,分别为对照组、低氟组（含25mg/L NaF去离子水）、中氟组（含50mg/L NaF去离子水）、高氟组（含100mg/L NaF去离子水）,在围产期经水攻毒,待小鼠断奶后处死,摘取肾脏做石蜡组织切片,HE染色后观察肾脏的病理变化.研究结果表明：仔鼠接受氟暴露后,肾小球出血,肾小管上皮细胞水肿,排列紊乱,核固缩,并且随着染氟剂量的增加,病变程度也逐渐加重.     

还原型谷胱甘肽对西门塔尔牛精液低温保存效果的影响

本实验旨在探究还原型谷胱甘肽（GSH）对西门塔尔牛精液低温保存（0℃）的影响。在精液稀释液中分别添加0（对照组）、1、2、4、6 mmol/L的GSH,在0℃条件下保存24、72、120、168、216 h时检测精子活力、顶体完整率和质膜完整性等指标;保存72 h和168 h时检测精子的丙二醛（MDA）水平、三磷酸腺苷（ATP）含量、谷胱甘肽过氧化物酶（GSH-Px）和过氧化氢酶（CAT）活性。结果表明：在精液低温保存72 h时,4 mmol/L GSH添加组的精子活力、顶体完整率、质膜完整性、CAT活性、ATP含量和GSH-Px活性高于对照组（P<0.05）;在精液低温保存168 h时,4 mmol/L GSH添加组的精子活力、顶体完整率、质膜完整性、CAT活性、ATP含量和GSH-Px活性高于对照组与其他GSH添加组（P<0.05）,MDA水平低于其他组（P<0.05）。因此,在本实验条件下,在精液稀释液中添加4 mmol/L GSH可以有效提高精液品质和精子的抗氧化能力,延长精子的保存时间。     

褪黑素、谷胱甘肽对猪精液冷冻保存效果的影响

本试验旨在研究褪黑素、谷胱甘肽两种抗氧化剂对精子冷冻的保护效应。以成年杜洛克公猪为研究对象,在猪精液冷冻稀释液中先后单独、联合添加褪黑素、谷胱甘肽,解冻后精子质量通过检测活率、活力、顶体完整性、线粒体活性以及活性氧（ROS）含量来判定。结果表明：分别单独添加0.25mg/mL褪黑素、5mmol/L谷胱甘肽,或联合添加0.125mg/mL褪黑素和1mmol/L谷胱甘肽,均能减少精液冷冻过程中ROS的生成,显著提高冷冻精子解冻后的质量（P〈0.05）,其中联合添加效果显著优于单独添加效果。     

谷胱甘肽对犬精液冷冻保存效果的影响

为探讨在冷冻稀释液中添加谷胱甘肽（GSH）对犬精液冷冻保存效果的影响,采用按摩法采集5只杂种土犬的精液,离心去精清后,在冷冻液中分别加入0.5、1.0、1.5、2.0、2.5 mmol/L的GSH,制成0.25 mL的冻精进行冷冻保存,以不添加GSH的处理组作为对照组。解冻后在含有5% CO₂的空气、37 ℃、相对饱和湿度条件下孵育10 h,分别在孵育0、2、4、6、8、10 h时检查精子活力。结果显示：冻融后0 h,0.5、1.0 mmol/L GSH处理组的精子活力较高,分别为0.36和0.38,均显著（P<0.05）高于对照组和2.0、2.5 mmol/L处理组,且两者之间差异不显著（P>0.05）;1.0 mmol/L处理组的精子顶体完整率最高,为85.10%,显著（P<0.05）高于对照组,同时,其精子畸形率最低,为23.00%,显著（P<0.05）低于对照组。冻融后体外孵育2、4 h时,0.5、1.0 mmol/L处理组的精子活力均较高,其中,0.5 mmol/L处理组在体外孵育4 h时,其精子活力仍可达到0.30;孵育6 h时,1.0 mmol/L处理组精子活力最高,显著（P<0.05）高于对照组和2.0、2.5 mmol/L处理组;孵育8 h时,各GSH处理组的精子活力均显著（P<0.05）高于对照组;在孵育至10 h时,各GSH处理组的精子活力较其他孵育时间均有较大幅度的下降,未检测到对照组中有呈直线运动的精子。综上提示,在犬精液冷冻液中添加0.5~1.0 mmol/L的GSH能够显著提高冻融后的精子质量和体外存活时间。     

氟对小鼠附睾中成熟精子超微结构的影响

本试验探讨了氟对小鼠附睾中成熟精子超微结构的影响,为氟的生殖毒性研究与检测提供依据。选取8周龄性成熟雄性昆明小鼠20只,随机分为4组,对照组小鼠饮用蒸馏水,低、中和高氟组小鼠分别饮用含25、50和100 mg/L氟化钠的蒸馏水,于45 d后断颈处死小鼠,取小鼠附睾尾,经2.5%戊二醛固定后,采用透射电镜观察小鼠附睾中成熟精子的超微结构变化。与对照组相比,低氟组小鼠精子头部质膜断裂,部分脱落,个别线粒体肿胀、形态模糊;中氟组精子头部质膜脱落,顶体部分缺失,线粒体形状不规则,嵴间腔扩大;高氟组精子头部质膜脱落,线粒体排列不规则,出现空泡化,嵴结构模糊。结果表明,氟暴露小鼠附睾中成熟精子头、尾部中段均有不同程度的结构改变,尤以线粒体出现较明显的异常,且氟浓度越高,精子超微结构损伤越严重。     

莱菔硫烷对镉中毒雄性小鼠生殖机能保护作用的研究

本试验旨在研究莱菔硫烷(SFN)对染镉雄性小鼠生殖机能的保护作用.选择健康的清洁级雄性昆明系小鼠40只,随机分成4组,分别为对照组(H₂O)、镉组(2.3 mg/kg CdCl₂)、莱菔硫烷组(10 mg/kg SFN)、镉+莱菔硫烷组(10 mg/kg SFN+2.3 mg/kg CdCl₂),连续给药10 d,于最后一次给药2 d后脱颈处死,检测小鼠睾丸组织病理学变化、小鼠睾丸及附睾的脏器系数、精液品质及血清睾酮浓度.同时检测睾丸组织内丙二醛(MDA)、谷胱甘肽(GSH)含量及总超氧化物歧化酶(T-SOD)活性的变化.结果表明,与对照组相比,镉组小鼠的睾丸组织发生病理学损伤,睾丸及附睾的脏器系数、精液品质及血清睾酮浓度均极显著降低(P<0.01),睾丸组织T-SOD活性、GSH含量也极显著降低(P<0.01),MDA含量极显著提高(P<0.01);莱菔硫烷组T-SOD活性和GSH含量显著升高(P<0.05),MDA含量与对照组相比差异不显著(P>0.05).与镉组相比,镉+莱菔硫烷组小鼠的精子活率、精子总数极显著增加(P<0.01),睾丸与附睾的脏器系数显著增加(P<0.05),小鼠睾丸组织GSH含量、T-SOD活性极显著提高(P<0.01),MDA含量极显著降低(P<0.01).以上结果指出莱菔硫烷对镉中毒雄性小鼠的生殖机能具有保护作用.     

Protective Effect of Sulforaphane on Reproductive Function of Male Mice with Cadmium Poisoning

The study was aimed to explore the protective effect of sulforaphane (SFN) on the reproductive function of male mice with cadmium poisoning.40 healthy clean grade male Kunming mice were randomly divided into four groups:control group (H₂O),cadmium chloride group (2.3 mg/kg CdCl₂),sulforaphane group (10 mg/kg SFN),sulforaphane + cadmium chloride group (10 mg/kg SFN+2.3 mg/kg CdCl₂),and continuous administration for 10 d,all mice were executed by dislocated cervical vertebra at 2 d after the last administration,and then the pathologic changes of testicular tissues,organ coefficient of testicle and epididymis,sperm quality and concentration of testosterone were tested.Additionally,the contents of GSH and MDA,and the activities of T-SOD in testis were also detected at the same time. Compared with the control group,pathology damages were observed in cadmium chloride group,organ coefficient of testis and epididymis,sperm quality and levels of testosterone extremely significantly decreased (P<0.01),the activities of T-SOD and GSH content were extremely significantly decreased (P<0.01),and the concentration of MDA was extremely significantly enhanced (P<0.01).Compared with the control group,the activity of T-SOD and concentration of GSH in sulforaphane group were significantly increased (P<0.05),and the concentration of MDA was not significant different between the control group and sulforaphane group (P>0.05).While compared with the cadmium chloride group,the sperm motility rate and sperm total count in sulforaphane and cadmium chloride group were extremely significantly increased (P<0.01),the organ coefficient of testicle and epididymis was increased significantly (P<0.05),the concentration of GSH and activity of T-SOD in testicular tissue were extremely significantly increased (P<0.01),and the concentration of MDA was extremely significantly decreased (P<0.01).The results indicated that sulforaphane had the protection effect on reproduction function of male mice with cadmium poisoning.     

Effects of Sodium Fluoride on the Morphological Structure of the Epididymis and the Expression of Tight Junction Genes

The purpose of this study was to examine the effects of fluoride on epididymal morphological structure and mRNA expression of tight junction genes.60 healthy male mice were divided into four groups randomly, including control group (distilled water) and 25,50,100 mg NaF/L groups, respectively. After 60 days, the thighbones were collected and fluoride contents were determined by fluorine ion selective electrode method. Paraffin slides of right epididymis with HE stain were produced to evaluate epididymal morphological structure under light microscope. The left epididymis were collected to isolate total RNA and detect the mRNA expression of tight junction genes by qRT-PCR. The results showed that the fluoride contents of thighbones increased in a dose-dependent manner, and were extremely significantly higher in 50 and 100 mg/L groups than those in control group (P<0.01).NaF extremely significantly increased the thickness of cauda epididymal pipe wall in fluoride groups (P<0.01) and the Claudin-2 mRNA expressions in fluoride groups were extremely significantly decreased (P<0.01) and the ZO-1 mRNA expression in 100 mg/L group was significantly decreased (P<0.05) as compared with control group. In conclusion, excessive NaF ingestion adversely affected epididymal morphological structure and mRNA expressions of Claudin-2 and>ZO-1,which might contribute to the damage of blood-epididymal barrier.     

氟对雄性小鼠生长发育及性腺中氟含量的影响

本试验旨在研究氟对性成熟期雄性小鼠生长发育及性腺中氟含量的影响。将40只健康昆明雄性小鼠随机分为4组,即低、中、高3个染氟组和对照组,其中染氟组分别饮水摄入氟化钠30、70和150 mg/L,对照组饮用蒸馏水,染氟期间,观察动物的一般状况,每周记录小鼠体重,49 d后,收集血液、股骨、睾丸、附睾和精囊腺,测定其氟含量。与对照组相比,各染氟组体重均无显著差异(P＞0.05),高氟组小鼠体增重显著低于对照组(P＜0.05);各染氟组股骨氟含量均显著高于对照组(P＜0.05),睾丸和附睾中氟含量随摄氟剂量的增加呈剂量依赖性,且高氟组显著升高(P＜0.05),血清与精囊腺中氟含量与对照组相比无显著差异(P＞0.05)。结果表明,高氟对性成熟期雄性小鼠生长发育有抑制作用,且突破了血睾屏障和血-附睾屏障进而对生殖系统产生影响。     

不同硒浓度日粮对小鼠肝脏和睾丸组织中部分硒蛋白mRNA水平的影响

选取80只7周龄雄性BALB/c小鼠,随机分为Ⅰ、Ⅱ、Ⅲ、Ⅳ组,每组20只,使Ⅰ、Ⅱ、Ⅲ、Ⅳ组基础日粮中硒含量达到0.045、0.1、0.4和0.8 mg/kg。结果表明:①Ⅰ组能显著降低小鼠肝脏中硒的沉积量(P<0.05);②Ⅰ组谷胱甘肽过氧化物酶1(glutathione peroxidase 1,GPx1)的mRNA相对表达量明显低于Ⅱ、Ⅲ组(P<0.05),且Ⅱ、Ⅲ组在肝脏中差异显著(P<0.05),在睾丸中差异不显著(P>0.05);③Ⅰ、Ⅲ组肝脏中谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPx4)的mRNA相对表达量差异不显著(P>0.05),但均显著低于Ⅱ组(P<0.05),而各试验组间在睾丸中差异均不显著(P>0.05);④随着日粮中硒添加量的增加,肝脏和睾丸中硫氧还蛋白还原酶2(thioredoxin reductase 2,TrxR2)的mRNA相对表达量显著提高(P<0.05)。因此,在肝脏和睾丸组织中,饲喂低硒饲料能显著降低硒的沉积量和GPx1、GPx4、TrxR2的mRNA相对表达量(P<0.05);而饲喂高硒饲料能显著降低GPx1的mRNA相对表达量(P<0.05),同时显著增加TrxR2的mRNA相对表达量(P<0.05)。提示,硒对GPx1、GPx4和TrxR2 mRNA相对表达量的调节存在组织差异性。     

Effect of Luteolin on Blood Indexes,Liver and Kidney in Mice with Acute Mercury Poisoning

To study the effect of luteolin on blood indexes,liver and kidney in mice with acute mercury poisoning,28 mice were randomly divided into four groups:Control group (intraperitoneal injection 0.9% saline),luteolin group (lavage 100 mg/kg luteolin),mercuric chloride group (intraperitoneal injection 4 mg/kg mercury chloride) and mercury chloride+luteolin group (intraperitoneal injection 4 mg/kg mercuric chloride,lavage 100 mg/kg luteolin).The activities of ALT in serum,AST,CREA and BUN contents,blood WBC,RBC,HGB content and GSH and MDA contents of liver tissue were detected.Morphological changes of liver and kidney tissues were observed.The results showed that compared with the control group,the activities of ALT and AST of mercuric chloride group were extremely significantly increased (P < 0.01),serum CREA,BUN,blood WBC and liver tissue MDA contents were significantly increased (P < 0.05) while blood RBC,HGB and liver tissue GSH contents were significantly decreased (P < 0.05).Liver,kidney pathological changes were obviously.Compared with mercuric chloride group,the activities of ALT,AST in serum,CREA,BUN,blood WBC and liver tissue MDA contents of mercuric chloride+luteolin group were significantly decreased (P < 0.05),while blood RBC,HGB and liver tissue GSH contents were significantly elevated (P < 0.05).Liver,kidney pathological changes were attenuated obviously.The poisoning were characterized by inflammation and the occurrence of anemia when acute mercury poisoning occurred,liver and kidney showed different degrees of injury in mice.Luteolin could reduce the toxic effects of acute mercury poisoning on blood,liver and kidney.     

木犀草素对急性汞中毒小鼠血液生化指标、肝脏及肾脏的影响

试验旨在研究小鼠急性汞中毒后血液、肝脏及肾脏的中毒表现及木犀草素对急性汞中毒小鼠血液指标、肝脏及肾脏的影响。将28只小鼠随机分成4组,分别为对照组(腹腔注射0.9%生理盐水)、木犀草素组(灌胃100 mg/kg木犀草素)、氯化汞组(腹腔注射4 mg/kg氯化汞)和氯化汞+木犀草素组(腹腔注射4 mg/kg氯化汞,灌胃100 mg/kg木犀草素)。检测血清谷丙转氨酶(ALT)和谷草转氨酶(AST)活性,肌酐(CREA)和尿素氮(BUN)水平,血液白细胞(WBC)、红细胞(RBC)和血红蛋白(HGB)水平及肝脏组织谷胱甘肽(GSH)和丙二醛(MDA)水平,并观察肝脏、肾脏组织形态学改变。结果显示,与对照组相比,氯化汞组的血清ALT、AST活性极显著升高(P < 0.01),血清CREA、BUN,血液WBC及肝脏组织MDA含量均显著增加(P < 0.05),血液RBC、HGB及肝脏组织GSH水平均显著降低(P < 0.05),肝脏、肾脏病理变化明显。与氯化汞组相比,氯化汞+木犀草素组的血清ALT、AST活性,CREA和BUN,血液WBC及肝脏组织MDA含量均显著降低(P < 0.05);血液RBC、HGB及肝脏组织GSH含量均显著升高(P < 0.05),肝脏、肾脏病理变化减轻。小鼠急性汞中毒后,其中毒表现为炎症和贫血的发生及肝脏、肾脏不同程度的损伤,木犀草素可减弱急性汞中毒小鼠血液、肝脏和肾脏的毒性作用。