【作者】 李升和；

【导师】 彭克美；

【作者基本信息】 华中农业大学 ， 预防兽医学， 2008， 博士

【摘要】 硼(B)是地壳中含量很低的一种非金属微量元素,是植物的必需微量元素。硼及其化合物广泛用于工业、农业、畜牧业、化妆品和医疗卫生等部门。近年来研究表明,适量硼对人和动物具有营养作用,高剂量硼则对人和动物具有不利影响,甚至毒性作用。本课题以196只清洁级SD大鼠为试验动物,研究饮水添加硼40、80、160、320、640mg/L对大鼠的生长与器官发育、血液生理生化、血清和器官抗氧化能力、以及舌、胃、肝脏、胰腺、睾丸、附睾、脾脏和腺垂体等器官显微和超微结构的影响,探讨硼对大鼠的营养和毒性作用机理。具体内容如下:1.硼对大鼠生长及器官发育的影响通过测定大鼠体重、体长和器官质量或长度,计算Lee’s指数和相应器官指数,研究硼对雌雄大鼠生长和器官发育的影响。结果显示:饮水添加硼40～320mg/L对大鼠BM、ADG、Lee’s指数、器官质量及肠管长度与指数多无显著影响(p＞0.05),对65d雄性大鼠肝、肾上腺、胸腺和睾丸质量有多有显著降低作用(p＜0.05或p＜0.01);添加硼40～80mg/L对95d大鼠器官质量多有升高作用,但差异不显著(p＞0.05);添加硼640mg/L对大鼠BM、ADG、Lee’s指数和器官质量(或长度)及其指数多有显著影响(p＜0.05或p＜0.01)。结果表明,饮水添加硼40～80 mg/L对65d大鼠生长及器官发育有较明显促进作用;添加硼160～320mg/L多有较明显的不良影响;添加硼640mg/L则有明显的抑制甚至毒性作用。2.硼对大鼠血液生理生化的影响通过血细胞计数、血红蛋白含量测定、血清部分生化指标测定、血涂片染色及血细胞形态结构观察,研究硼对大鼠血液生理生化的影响。结果显示:与同日龄对照组相比较,各试验组网织红细胞数不同程度升高;试验Ⅰ、Ⅱ组红细胞数和血红蛋白含量多显著升高(P＜0.05),白细胞数和白细胞分类计数多无显著差异(P＞0.05);试验Ⅲ～Ⅴ组的红、白细胞数和血红蛋白含量显著降低(P＜0.05或P＜0.01)低于同日龄对照组,中性粒细胞、嗜酸性粒细胞、嗜碱性粒细胞和单核细胞比例多显著降低(P＜0.05或P＜0.01),95d淋巴细胞比例显著升高(P＜0.05)。试验Ⅰ组红细胞和无粒白细胞形态结构无明显变化,有粒白细胞胞核体积增大;试验Ⅱ～Ⅴ组,随硼添加量增大,变形、破损、崩解的红细胞增多,淋巴细胞胞核先固缩后肿胀,有粒白细胞胞质内颗粒减少或消失,胞核难以着色,试验Ⅴ组多叶型中性粒细胞增多。试验Ⅰ～Ⅳ组血清胆固醇含量不同程度降低,试验Ⅴ组显著升高(P＜0.05或P＜0.01)。65d试验Ⅲ、Ⅴ组和95d试验Ⅴ组血糖水平显著降低(P＜0.05或P＜0.01)。各试验组血清TRP含量差异不显著(P＞0.05)。65d试验Ⅲ组和95d试验Ⅴ组血清ALT活性显著降低(P＜0.05)。65d试验Ⅴ组血清AST活性显著升高(P＜0.05),95d试验Ⅰ、Ⅴ组显著降低(P＜0.05)。65d试验Ⅴ组血清ALP活性显著降低(P＜0.05)。65d试验Ⅰ组血钙水平显著升高(P＜0.05),65试验Ⅳ、Ⅴ组和95d试验Ⅱ～Ⅳ组显著降低(P＜0.05或P＜0.01)。95d试验Ⅳ、Ⅴ组血磷水平显著降低(P＜0.05)。65d试验Ⅲ组和95d试验Ⅴ组血镁水平显著下降(P＜0.05)。结果表明,饮水添加40mg/L硼对大鼠血细胞发生、增殖分化及形态结构与功能有明显促进作用;添加80～640mg/L硼对血细胞形态结构与功能有不同程度损伤,甚至毒性作用;饮水添加40～640mg/L硼对血清各项生化指标有不同程度的影响。3.硼对大鼠血清和器官抗氧化能力的影响使用SOD、CAT、MDA、GSH-PX和考马斯亮兰试剂盒测定血清和器官的SOD、CAT和GSH-PX活性,以及MDA和组织蛋白含量。结果显示:饮水添加40mg/L硼对大鼠血清各抗氧化指标无显著影响(P＞0.05);使95d多个器官SOD、CAT和GSH-PX活性有显著升高,部分器官MDA含量显著降低(P＜0.05或P＜0.01)。添加80mg/L硼可显著升高95d大鼠血清SOD活性及部分器官SOD、CAT和GSH-PX活性(P＜0.05或P＜0.01);显著降低延髓和肝MDA含量(P＜0.05)。添加160mg/L硼对95d大鼠血清SOD和部分器官SOD、CAT、GSH-PX活性,以及少数器官MDA含量有显著升高作用;对95d血清MDA含量及部分器官MDA含量、SOD和GSH-PX活性则有显著降低作用(P＜0.05或P＜0.01)。添加320mg/L硼对65d大鼠血清MDA含量,95d大鼠血清和部分器官SOD活性,多个器官MDA含量,以及部分器官CAT和GSH-PX活性有显著升高作用(P＜0.05或P＜0.01);对95d血清和多个器官GSH-PX活性,部分SOD和CAT活性有显著降低作用(P＜0.05或P＜0.01)。添加640mg/L硼对大鼠血清MDA含量及SOD、CAT和GSH-PX活性多有显著影响,多数器官SOD、CAT和GSH-PX活性显著降低(P＜0.05或P＜0.01);对95d大鼠各器官MDA含量有明显升高作用。结果表明,饮水添加40～80mg/L硼可不同程度提高机体的抗氧化能力;添加320～640mg/L硼对机体的抗氧化系统则有不良影响甚至毒性作用。4.硼对大鼠器官显微和超微结构的影响利用光镜和电镜制片技术观察95d大鼠舌、胃、肝、睾丸、脾和腺垂体等6个器官的显微和超微结构。结果显示:试验Ⅰ、Ⅱ组各器官组织结构发育良好,器官结构清晰,各种细胞排列规则、整齐;细胞内胞核场染色质增多,胞质内线粒体、RER和高尔基复合体等细胞器发达,腺垂体分泌细胞体积增大,分泌颗粒数量增多;各器官和细胞的结构呈现功能旺盛状态。表明饮水添加40～80mg/L硼对大鼠各器官的组织结构与功能有较明显的促进与增强作用,其中以添加40mg/L硼组的作用效果最好。试验Ⅲ组大鼠各器官组织结构可见不同程度轻微病变,表明饮水添加160mg/L硼对大鼠舌、肝脏、胰腺、睾丸等器官发育及组织结构与功能有较轻微不良影响或损伤作用。试验Ⅳ、Ⅴ组大鼠各器官显微和超微病变明显,舌丝状乳头顶部溶解,味蕾萎缩;胃黏膜上皮细胞坏死、脱落,胃底腺萎缩、结构混乱;肝细胞萎缩或肿胀,各种细胞器数量减少、结构不同程度破坏;睾丸萎缩,各种生精细胞显著减少,细胞内线粒体、高尔基复合体等细胞器破坏严重。各器官病理组织学变化与硼添加量存在明显相关性,表明饮水添加320和640mg/L硼对大鼠各器官发育及组织结构与功能有明显不良影响和损伤作用,甚至表现明显毒性作用。更多还原

【Abstract】 Boron,a kind of nonmetal element,with low content in crust.Boron has been used in industry,agriculture,animal husbandry,cosmetics,medical treatment and some other fields.It was showed that the quantum vis boron has trophic action on human beings and animals during the recent years.It was indicated by some animal experiment,high dosage of boron was bad for growth,development,metabolism and some other physiological function of men and animals,and even toxic function.In this experiment,192 rats,with 0, 40,80,160,320,640 mg/L boron in the drinking water,and the effects of boron on the growth and organ development of some organs,the blood physiology and biochemistry, the ability of anti-oxidation of serum and some organs,and the microstructure and ultrastrcture of tongue,stomach,liver,pancreas,sperm,parorchis,testicle spleen adenohypophysis were studied.At same time,it was told mechanism of action of boron nutrition and toxic function on microstructure and ultrastrcture level.And in this article, there was a certain dose range of boron nutrition and toxic function for rats initially.The concrete result was showed:1.Influence of growth and allelotaxy in rat by added to boronThe growth and allelotaxy were studied by detection of body weight,stem length and organic quality or length and calculation of Lee’s and corresponding organ index. The results showed by added 40～320mg/L boron to drinking water,the BM、ADG、Lee’s index、organ quality and intestinal length and index was not evidently influenced （p＞0.05） in rats,but there was a evident low in the quality of liver、adrenal gland、thymus and testicle of 65-days male rats（p＜0.05或p＜0.01）.By added 40～80mg/L boron to drinking water,the most of organic quality in 95-days rats were not evidently rose（p＞0.05）.By added 640mg/L boron to drinking water,the BM、ADG、Lee’s index、organ quality and intestinal length and index was evidently influenced in rats（p＜0.05或p＜0.01）.The results indicated that the growth and allelotaxy of rat were evidently promoted by added 40～80mg/L boron to drinking water,and evidently effected by added 160～320mg/L boron,and evidently repressed and even poisoned by added 640mg/L boron.2.Influence of physiological and biochemical indicator in rat by added to boronInfluence of physiological and biochemical were studied by blood cell count, hemoglobin level dectection,partial serum biochemical indicator dectection,blood smear and haemocyte morphological observeation.The results showed:compared with control group of same age,the reticulocytes percentage of each experiment group exhibited different degree increasing,the RBC numbers and Hb content of experiment groupⅠandⅡmore raised significantly（P＜0.05）,but the changes of the leucocytes percentage were more no significant（P＞0.05）.The numbers of RBC and WBC,the content of Hb and the percentage of neutrophil,eosinophil and monocytic in experimental group fromⅢtoⅤdecreased significantly or most significantly（p＜0.05 or p＜0.01）.Under the microscope, there were no obvious changers of all kinds of blood cells in experiment groupⅠ.In experiment group fromⅡtoⅤ,with the boron supplementation increasing,the deformed and disintegrated grew in number.The pyknotic nuclei of lymphocyte could be observed,The granulations in granular leukocyte decreased obviously or even vanished, and the nuclei in them were stained hardly,especially in experiment groupⅤ,some more segmented neutrophil with could be observed.The content of serum glucose in the experiment group was evidently lower than the control group on 65^th day,in which existed evident different in theⅢandⅤexperiment group（P＜0.05或P＜0.01）,the content of serum glucose in theⅤexperiment group was evidently lower than the control group on 95^th day（P＜0.05）.The activity of serum ALT in theⅢexperiment group with 65-days and 95-days was evidently lower than the control group（P＜0.05）.The activity of serum AST was evidently raise in theⅤexperiment group with 65-days（P＜0.05）, but lowed in theⅠandⅤexperiment group with 95-days（P＜0.05）.The activity of serum ALP was evidently lowed in theⅤexperiment group with 65-days（P＜0.05）.The content of serum Ca²⁺ was evidently rose in theⅤexperiment group with 65-days,but evidently or extremely lowed in theⅣandⅤexperiment group with 65-days andⅡ～Ⅳexperiment group with 95-days（P＜0.05或P＜0.01）.The content of serum phosphorus was evidently lowed in theⅣandⅤexperiment group with 95-days（P＜0.05）.The content of serum Mg²⁺ in theⅢexperiment group with 65-days andⅤexperiment group with 95-days was evidently lowed than the control group（P＜0.05）.The results indicated that biochemical indicators serum were influenced to different degree by added 40～640mg/L boron to drinking water.3.Influence of Boron on the ability of rat’s serum and organs’ anti-oxygenThe activity of SOD、CAT and GSH-PX was dectected by Coomassie light blue kit.The results showed it has no significant influence on each index of serum’s anti-oxygen of rat by added 40mg/L boron into drinking water（P＞0.05）.But activity of SOD,CAT and GSH-PX increased significantly of many organs of 95ds rat.Added 80 mg/L Boron,the activity of serum and SOD,CAT and GSH-PX of organs advanced notably on 95d,and the content of MDA of myelencephalon and liver depress significantly.Adding 160 mg/L Boron,activity of SOD of serum,hypothalamus,liver and pancreas,activity of CAT,GSH-PX of kidney and the content of MDA of pancreas and epididymal increase significantly.Content of MDA loss of 95d’s rat’s serum, hypothalamus and liver,SOD activity depress of myelencephalon and epididymis,and GSH-PX activity depress notably of myelencephalon,heart,lung and lymph node.Added 320 mg/L Boron,MDA content of serum of 65d’s rat increases significantly,SOD activity of serum,liver,kidney,pancreas and spleen of 95d increase notably,many organs’ MDA content and some organs’ CAT and GSH-PX activity increase significantly.95d rat’ serum and many organs’ GSH-PX activity,SOD activity of myelencephalon and epididymal, CAT activity of liver and epididymal,depress notably.Added 640 mg/L Boron,it had significant or extreme significant influence of MDA content of serum and activities of SOD,CAT and GSH-PX of 65d and 95d rats.It had notably effect on 95d rat’s organs content of MDA.Many organs’ activities of SOD,CAT and GSH-PX lower than control group notably.The results indicated the anti-oxygen ability of body was enchanced to different degree by added 40～80mg/L boron to drinking water,but effected and even prisoned by added 320～640mg/L boron to drinking water.4.Effect of Boron on the microstructure and ultrastructure of rat’s organsthe microstructure and ultrastructure of tongue,stomach,liver,testicle,spleen and adenohypophysis in 95d rat’s organs show that:inⅠandⅡexperimental groups, observation from light microscope,evey organ and tissue grow well,structures of each organ was clarity,cells arrange in rule.In cells,some organel such as Mitochondria,RER and golgi complex,were developed well,volume of extre cell in denohypophysis increased,number of secretory granules mainfolded,with high density.It showed us that appropriate content of Boron in drinking water can improved the structure and funciton of some organs obviously,when added 40mg/l,the effect was best.InⅢexperimental groups,it had some harmful or injury influence on the microstructure and ultrastructure of tongue,liver,pancreas and testicle in rat’s organs,There was obvious damage effect on microstructure and ultrastructure of rat organs inⅣandⅤexperimental groups,It showed filiform papillae top of tongue dissolved,taste goblet shrunk back;stomach mucous membrane epithelium was necrosis and desquamated,fundic glands turned shrink back,structure in a mess;hepatocytes turned analosis or tumentia,the cell organs in cytoplasm decreased,and the structure was damaged in different degree,it showed orchiatrophy,the number of spermatogenic cells in contorted seminiferous tubules decreased significantly,epithelium on tubal wall was damaged terribly,and cell organs, such as mitochondria and golgi complex,etc.All those changes showed that,there would be bad and even toxic effect on development and structure of every organ in rat by supplement 320-640 mg/L boron in drinking water,and influenced function of organism and all organs terribly.更多还原