应用流式细胞分析技术研究微纳尺度SiO2对雄性大鼠睾丸生精细胞的毒性作用

为了进一步研究纳米与微米尺度SiO2对雄性大鼠的生殖毒性作用,选择不同剂量的纳米SiO2(20～40nm)与微米SiO2(1～10μm),采用气管滴注方式对雄性Wistar大鼠分组染毒.于染毒5周后处死大鼠,应用流式细胞技术对睾丸生精细胞进行分析.结果表明:1)高、低剂量纳米SiO2组及高剂量微米SiO2组细胞凋亡率均显著高于对照组;2)与对照组相比,高、低剂量纳米SiO2组及高剂量微米SiO2组1C细胞显著减少,4C细胞显著增加;3)与对照组相比,高剂量纳米SiO2组和高剂量微米SiO2组G0/G1期细胞比例显著降低,高剂量纳米SiO2组G2/M期细胞比例显著增加.结果提示纳米SiO2能够阻滞细胞周期进程,诱导生精细胞凋亡;与微米SiO2相比,纳米SiO2对大鼠睾丸生精细胞的损伤有更严重的趋势.     

五氯酚钠对雄性小鼠的生殖毒性作用

为研究五氯酚钠(PCP-Na)对雄性小鼠的生殖毒性作用,将100只昆明雄性小鼠随机分为5组:3个染毒组、1个阴性对照组(蒸馏水)和1个阳性对照组(环磷酰胺40mg·kg-1).以不同剂量(13.44、26.88、53.76mg·kg-1)PCP-Na经口染毒,连续染毒5d,每天1次.染毒结束后第2d和第23d分两批处死动物,检测精子数量、精子畸形率以及睾丸、附睾重量及其脏器系数的变化.结果表明,PCP-Na染毒结束后第2d,与阴性对照组相比,高剂量组(53.76mg·kg-1)精子畸形率显著增加(p<0.05),睾丸、附睾重量及附睾系数显著下降(p<0.05),其他指标无显著变化(p>0.05).中、低剂量组(26.88、13.44mg·kg-1)各检测指标与阴性对照组均无显著性差异(p>0.05);PCP-Na染毒结束后第23d,与阴性对照组相比,高剂量组(53.76mg·kg-1)精子畸形率显著增加、精子密度显著减少、睾丸和附睾重量显著下降(p<0.05),中剂量组(26.88mg·kg-1)附睾重量显著下降(p<0.05),其他指标均无显著变化(p>0.05);PCP-Na引起的精子畸形以头部畸形为主,主要为无定形,其次为无钩形、香蕉形和尾折叠.以上结果提示,在本实验条件下,PCP-Na对雄性小鼠具有明显的生殖毒性.     

载带苯并[a]芘的纳米碳／二氧化硅颗粒引起大鼠氧化应激作用的研究

通过人工制备载带B[a]P的纳米碳(C)和纳米二氧化硅(SiO2)颗粒,采用气管滴注染毒方式,以7.5mg·kg-1(以体重计)的染毒剂量急性染毒大鼠,观察染毒24小时后载带B[a]P的纳米C/SiO2颗粒对机体产生氧化应激损伤的联合毒性效应.结果表明,在急性染毒后大鼠外周血中反映机体脂质过氧化损伤程度指标的丙二醛(MDA)含量表现为染毒组较对照组显著增加(p<0.05),表明纳米颗粒诱发机体发生了氧化应激反应.在急性染毒后各组大鼠肺泡灌洗液中谷胱苷肽过氧化物酶(GSH-PX)和超氧化物岐化酶(T-SOD)活力与对照组相比显著增加(p<0.05)(载带B[a]P的纳米SiO2组除外);载带B[a]P的纳米SiO2组肺泡灌洗液中GSH-PX活力与对照组相比无显著差异,而与单纯纳米SiO2组和B[a]P组比较显著降低,推测与抗氧化酶的一过性增高有关;载带B[a]P的纳米C组肺泡灌洗液T-SOD活力与其单纯纳米C组和单纯B[a]P组比较显著增加(p<0.05),由此表明载带B[a]P的复合纳米C/SiO2颗粒在致机体氧化损伤效应方面二者存在一定的协同作用.     

非暴露式气管滴注3种典型纳米材料对大鼠肝、肾的毒性效应

为探讨纳米二氧化硅(Nano-SiO2)、纳米四氧化三铁(Nano-Fe3O4)和单壁碳纳米管(SWCNTs)对大鼠肝、肾的毒性效应,将49只雄性Wistar大鼠随机分为7组,包括生理盐水对照组,以及3种纳米材料的低剂量组(2mg·mL-1)和高剂量组(10mg·mL-1),采用非暴露式气管滴注法染毒,每2d染毒1次,每次每只0.2mL,共染毒5周,眼眶取血后处死大鼠,称肝、肾重量计算脏器系数,测定大鼠血清中反映肝、肾功能的生化指标,并对肝、肾进行病理学观察.结果表明,1)3种纳米材料均可导致大鼠体重明显降低,但对肝、肾脏器系数无明显影响;2)病理学观察发现,3种纳米材料均可导致大鼠肝细胞轻度脂肪变性,肾脏则无明显改变;3)3种纳米材料均可导致大鼠肝功能异常,部分肝功能指标如谷草转氨酶(AST)、碱性磷酸酶(ALP)、谷丙转氨酶(ALT)等出现显著降低;4)3种纳米材料均可导致大鼠肾功能异常,部分肾功能指标如尿酸(UA)、肌酐(CREA)、尿素氮(BUN)等出现显著升高或降低.以上结果提示,经呼吸道染毒的Nano-SiO2、Nano-Fe3O4和SWCNTs均可对大鼠肝、肾产生一定的毒性效应.     

PFOS青春期暴露对成年期雄性大鼠的生殖毒性

为探究青春期的PFOS暴露对成年后的SD大鼠的生殖毒性,对出生后第21天(PND21)的SD大鼠经口灌胃不同剂量的PFOS(5、10和20mg·kg~(-1)),连续染毒7d,在出生后第56天(PND56)时,对各染毒组SD大鼠的体质量、精子数量、血清中的睾酮浓度,以及睾丸间质细胞睾酮合成的相关基因mRNA水平进行了检测。结果显示,10mg·kg~(-1)剂量组大鼠体质量较对照组明显下降(P<0.01);精子数量在10mg·kg~(-1)和20mg·kg~(-1)剂量组明显降低(P<0.05);血清中睾酮浓度随着PFOS剂量的加大有明显下降的趋势,20mg·kg~(-1)剂量组显著低于对照组(P<0.05);类/胆固醇相关基因star和cyp11α1的mRNA表达水平明显下调。研究表明,青春期的PFOS暴露会导致睾酮合成途径中相关因子的功能缺失,破坏成熟睾丸间质细胞的功能,致使睾酮水平降低,并抑制精子生成,从而破坏生殖系统的功能。     

邻苯二甲酸丁基苄酯对小鼠睾丸能量代谢相关酶的影响

为了从生殖细胞能量代谢角度探讨邻苯二甲酸丁基苄酯(BBP)对雄性小鼠生殖损伤的机制,以昆明系雄性小白鼠为实验对象,研究了BBP对小鼠睾丸组织能量代谢相关酶的影响.实验设0、125、250、500、1000mg·kg-1(质量分数)5组浓度梯度,连续灌胃染毒30d后,采用分光光度法检测小鼠睾丸组织匀浆中乳酸脱氢酶(LDH)、琥珀酸脱氢酶(SDH)、Ca-Mg-ATPase酶的活性以及睾丸脏器系数.结果发现,1)与对照组相比,高浓度染毒组(500、1000mg·kg-1)LDH、SDH和Ca-Mg-ATPase活性均极显著降低(p<0.01);而低浓度染毒组(125mg·kg-1)3种酶活性均无显著变化(p>0.05);中浓度染毒组(250mg·kg-1)LDH没有显著变化(p>0.05),而SDH、Ca-Mg-ATPase均显著降低(p<0.05,p<0.01).2)染毒30d后,各染毒组睾丸脏器系数略有下降,但与对照组相比,差异均未达到显著水平(p>0.05).以上结果表明,较高水平的BBP不仅可以干扰睾丸组织有氧代谢及无氧代谢的产能过程,还可以干扰生殖细胞对能量的利用,提示能量代谢的障碍可能是BBP对雄性生殖细胞产生损伤的原因之一.     

二氧化硫体内衍生物对雄性小鼠精子的毒性效应

为了探讨二氧化硫(SO2)的雄性生殖毒理效应,研究了SO2体内代谢衍生物———亚硫酸钠(Na2SO3)和亚硫酸氢钠(NaHSO3)混合液(物质的量比为3:1)对雄性昆明小鼠精子的毒性作用.采用腹腔注射(i.p.)染毒方法,每天注射一次,连续5 d,于染毒后d 35处死动物,观察和分析小鼠的精子活动度、数量和畸形率.结果表明:(1)SO2衍生物可引起小鼠精子数量尤其是正常活动度精子数量减少,引起不活动精子数量增加,且呈剂量-效应关系;(2)SO2衍生物可引起多种类型的精子畸形增加,尤其可引起不定形畸变精子的频率显著增加,总精子畸变率随染毒剂量的增加而呈剂量依赖性增高.由此推论,SO2污染有一定的雄性生殖毒性.图1表2参14     

大气细颗粒物PM2.5对大鼠睾丸组织细胞周期的影响

为了研究大气细颗粒物(PM2.5)对雄性大鼠的生殖毒性作用,将不同剂量的PM2.5颗粒物悬液采用气管滴注方式对雄性Wistar大鼠分组染毒,染毒剂量分别为0、1.6、8.0、40.0mg·kg-(1BW),24h后应用流式细胞术对睾丸组织细胞周期及DNA倍体进行检测分析.结果显示:PM2.5对大鼠睾丸组织各倍体细胞比例和细胞周期有明显影响,可导致二倍体细胞数显著降低(p<0.05);可引起以二倍体细胞为主的G0/G1期细胞比例显著下降(p<0.05),G2/M期细胞比例和细胞增殖指数(PI)显著上升(p<0.05).结果提示,PM2.5可透过血睾屏障,干扰细胞周期进程,对生殖系统具有一定的毒性作用.     

铅与纳米SiO2联合染毒对A549细胞氧化损伤的影响

为了研究铅与纳米SiO2联合染毒所致的细胞损伤特征,并从氧化应激方面探讨其可能的作用机制。用铅和SiO2处理A549细胞,采用四唑盐(MTT)比色法检测细胞存活率,评价铅和SiO2联合染毒所致的细胞损伤特征;采用硫代巴比妥酸(TBA)比色法检测细胞内丙二醛(MDA)含量,评价铅与SiO2联合染毒所致细胞的氧化应激状态;检测了细胞内抗氧化物还原型谷胱甘肽(GSH)含量以及细胞内抗氧化酶的活性,以评价铅与SiO2联合染毒对细胞抗氧化系统的影响。将实验数据进行ANOVA分析。结果表明,铅、SiO2单独染毒组各指标没有明显改变;而联合染毒能造成细胞氧化损伤,表现为细胞存活率、GSH水平、超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GSH-Px)活性显著低于对照组及2个单独染毒组(P<0.05),细胞内MDA含量显著高于对照组及各单独染毒组(P<0.05)。可见,联合染毒可引起明显的细胞毒性,氧化损伤可能是铅与SiO联合染毒致肺细胞毒性损伤的作用机制之一。     

氰戊菊酯与辛硫磷混合杀虫剂（农满忆）对雄性小鼠精子畸形率和生殖细胞微核率的影响

为初步探讨氰戊菊酯与辛硫磷混合杀虫剂(农满忆)对雄性小鼠生殖细胞的损伤效应,将该杀虫剂对小鼠进行灌胃染毒5d,于第6d取小鼠睾丸细胞进行微核试验并采用精子畸形实验检测小鼠不同时期(第6、14、36d)精子畸形率的改变.结果表明,与对照组相比,该杀虫剂中、高剂量组(60、120mg·kg-1)小鼠睾丸细胞微核率均显著升高(p<0.05),而低剂量组(30mg·kg-1)微核率没有显著变化(p>0.05);中、高剂量组均能诱导小鼠不同时期精子畸形率显著升高(p<0.05),而低剂量组精子畸形率变化不显著(p>0.05).因此,氰戊菊酯与辛硫磷混合杀虫剂能够使雄性小鼠精子畸形率增加并造成雄性生殖细胞微核率升高,有一定的致突变作用.     

Protein expression profile in the testis of rats chronically exposed to low-dose 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin

The chronic effects low-dose 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) were examined on protein expression profiles in rat testis, sperm, and serum gonadal hormones. A total of 32 male rats were randomly divided into three TCDD-exposed groups, administered either 140, 350, or 875 ng TCDD/kg/week for 29 weeks, respectively, and one control group receiving only corn oil. The proteins from rat testis were separated and analyzed by two-dimensional gel electrophoresis and mass spectrometry. TCDD induced significant decreases in sperm counts and serum gonadal hormone levels compared with controls. TCDD altered testicular protein expression levels. Several interesting volume-altered proteins that were related to the reproductive toxicities or other toxicities of TCDD were identified. Among these proteins, PERF15 was the only down-regulated protein; sperm protein SSP411, ubiquitin carboxyl-terminal hydrolase L-3, and eukaryotic translation elongation factor 1 gamma were up-regulated by TCDD. The differentially expressed proteins and other data provide further insight into the mechanisms of reproductive toxicity mediated by low-dose TCDD exposure.     

Effect of malathion on reproductive system of male rats

The pesticides are one of the most potentially harmful chemicals liberated in the environment in an unplanned manner Malathion is widely used as a potent pesticide in many countries and has been shown to produce some adverse health effects. A study was conducted to asses the effects of malathion on the male reproductive system of wistar rats. The pesticide was administered to rats orally at dose levels of 50, 150 and 250 mg/kg/body wt/day for 60 days. In comparison to the control rats, there was a significant reduction in the weight of testes, epididymis, seminal vesicle and ventral prostate. Testicular and epididymal sperm density were decreased in the animals treated with malathion. Pre and post fertility test showed 80% negative results after treatment Biochemical profile of the testis revealed a significant decline in the contents of sialic acid and glycogen. Whereas a significant increase in the protein content of testis and testicular cholesterol was observed. The activity of testicular enzyme acid phosphatase increased significantly while decreased alkaline phosphatase activity was found. Malathion also suppressed the level of testosterone significantly Results of the present study clearly suggest that malathion induce toxic effects on the male reproductive system of rats.     

毒死蜱对雄性小鼠生殖毒性的影响

为研究毒死蜱对雄性小鼠生殖毒性的影响,利用不同浓度毒死蜱染毒小鼠.以昆明小鼠为受试动物,毒死蜱按3、6和12mg·kg-13个剂量水平,灌胃染毒小鼠7d.以睾丸组织匀浆测定活性氧(reactive oxygen species,ROS)和还原型谷胱甘肽(glutathione,GSH)的含量;以睾丸细胞测定DNA-蛋白...     

多壁碳纳米管引起白鼠肺部毒性的评价(Comparative Pulmonary Toxic Assessment of Mutil-Wall Carbon Nanotubes in Rats)

以雄性Wistar大白鼠为研究对象,观察了多壁碳纳米管(MWNTs)体内暴露后肺部的病理学变化.采用气管注入方式将直径为40～60nm多壁碳纳米管(40～60MWNTs)和直径小于10nm的多壁碳纳米管(10MWNTs)暴露于大白鼠的体内,同时分别采用纳米二氧化硅(SiO2)和乙炔碳黑(Cb)作为阳性和阴性对照.实验结果表明:实验中采用的纳米颗粒均不同程度地引起了肺部的损伤,同剂量下损伤严重程度顺序为:Cb>10MWNTs>40～60MWNTs>SiO2.多壁碳纳米管对肺部损伤的程度与剂量呈依赖关系,当在低剂量(1mg·kg-1)时,40～60MWNTs仅仅对肺部引起了轻微的炎症损伤,随着剂量的增大,多壁碳纳米管对肺部的损伤越来越严重,出现明显的病理损伤.     

纳米SiO2与常规SiO2颗粒对Hela细胞的细胞毒性作用

为了探讨纳米SiO2和常规SiO2颗粒对Hela细胞的细胞毒性作用,采用不同浓度的纳米SiO2和常规SiO2颗粒(0.05、0.1、0.2、0.4、0.8、1.6μg·μL-1)对Hela细胞进行12h染毒,应用MTT法检测细胞毒性效应.研究发现,较低浓度(≤0.2μg·μL-1)的纳米SiO2和常规SiO2对Hela细胞无明显细胞毒性(p>0.05);较高浓度时,纳米SiO2(≥0.4μg·μL-1)和常规SiO2(≥0.8μg·μL-1)对Hela细胞具有明显细胞毒性作用(p<0.01),并且随浓度增大细胞毒性增强;当浓度≥0.4μg·μL-1时,纳米SiO2的细胞毒性明显高于相同浓度的常规SiO2(p<0.05).以上结果表明,纳米SiO2和常规SiO2颗粒均能对Hela细胞产生细胞毒性,且纳米SiO2的细胞毒性强于常规SiO2;低浓度(≤0.2μg·μL-1)的纳米SiO2和常规SiO2具有很好的生物相容性.     

Protective effects of curcumin on antioxidant status,body weight gain,and reproductive parameters in male rats exposed to subchronic 2,3,7,8-tetrachlorodibenzo-p-dioxin

The aim of this study was to investigate the effects of curcumin (CUR) on antioxidant status, body weight (BW) gains, and some reproductive parameters in male rats exposed to subchronic doses of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Thirty-two rats were divided into four groups. The first group was kept as control. The second group (TCDD group) was given TCDD at a dose of 50 ng·kg^?1 BW per day; the third group (CUR group) was treated with CUR at a dose of 80 mg·kg^?1 BW per day. The fourth group (TCDD + CUR group) was given TCDD and CUR at the same doses simultaneously. Malondialdehyde (MDA) levels were significantly increased in the TCDD group. In addition, TCDD exposure decreased liver superoxide dismutase (SOD) activity, catalase (CAT) activities of kidney and brain, glutathione peroxidase (GSH-Px) activities of liver, kidney, and brain, and glutathione levels of liver, kidney, and heart. However, CUR treatment with TCDD exposure decreased MDA levels in all tissues and increased SOD activities of liver, kidney, and brain, CAT activity of heart, and GSH-Px activities of heart and brain. TCDD caused a decrease in BW gain, and CUR partially eliminated this effect of TCDD. In addition, while reproductive organ weights, sperm concentration, and sperm motility tended to decrease with TCDD exposure, these effects tended to be close to normal levels by CUR treatment. In conclusion, CUR was seen to be effective in the treatment and prevention of toxicity induced by subchronic TCDD exposure.     

Influence of hypothyroidism on oxidative stress,c-Fos expression,cell cycle,and apoptosis in rats testes

The molecular basis of male reproduction for cross-regulation between androgen and thyroid hormone axes is still rudimentary. This study aims to define a possible mechanism of hypothyroidism-induced reproductive influence with respect to sex hormone, mineral, sperm motility, oxidative stress, c-Fos expression, cell cycle, and apoptosis in rat testes. The Wistar rats were randomly divided into control group (NS) and hypothyroidism group [1 ml/100g BW/day, 0.1% propylthiouracil (PTU)] by intragastric gavage for 60 days. Blood samples were collected to measure the serum levels. The epididymis was excised to measure sperm motility and testes were excised to measure mineral, oxidative stress, c-Fos expression, cell cycle, and apoptosis. After 60 days, body weight, relative testes weight, triiodothyronine, and total thyroxine were all significantly decreased, whereas thyroid stimulating hormone was increased in the hypothyroidism group. A significant increase in sex hormone level of estradiol (E2) and significant decreases in testosterone (T) and T/E2 ratio were observed following PTU treatment. And sperm quality was also significantly changed. There were significant decreases in the contents of calcium (Ca²⁺) and zinc (Zn²⁺). On the other hand, malondialdehyde and hydrogen peroxide contents significantly increased, whereas the activity of superoxide dismutase, catalase and nitric oxide synthase and nitric oxide content significantly decreased in hypothyroid rats. The mRNA and protein expressions of c-Fos decreased significantly. The cell percentage in G0/G1 phase increased significantly, whereas decreased significantly in S and G2/M phases. Also, a significant increase in testicular cell apoptosis was observed in hypothyroid-treated rats. These results suggested that hypothyroidism could affect reproductive function in the form of changed sex hormone levels, sperm motility and testicular Ca²⁺ and Zn²⁺, and enhanced oxidative stress leading to c-Fos abnormal expression and increased apoptosis.