摘要
【目的】研究小花棘豆毒性成分对家兔睾丸、附睾组织中超氧化物歧化酶(superoxide dismutase,SOD)活性及基因表达的影响。【方法】将48只雄性家兔随机分为对照组和试验Ⅰ、Ⅱ、Ⅲ组,试验组分别按照Ⅰ组15%(苦马豆素含量30 mg/kg)、Ⅱ组30%(苦马豆素含量60 mg/kg)、Ⅲ组45%(苦马豆素含量90 mg/kg)的比例添加小花棘豆,对照组仅饲喂苜蓿干草,试验期70 d。分别于攻毒后第14、35、70 d每次每组随机采集4只家兔的睾丸和附睾组织,通过羟胺法检测SOD活性的变化,real-time PCR检测SOD mRNA的表达,免疫组化和Western Blot检测SOD蛋白的表达。【结果】从第35 d开始,与对照组相比,各试验组家兔附睾、睾丸中T-SOD、Cu/Zn-SOD和Mn-SOD活性均极显著下降(P﹤0.01),mRNA表达量和蛋白表达量均呈现出明显的下降趋势,而且其差异性随中毒时间的延长而变化。【结论】小花棘豆中毒可导致家兔睾丸、附睾中SOD活性与表达异常,且与小花棘豆中毒呈现一定的时间-剂量效应。
[Objective] The aim of this study was to investigate the effects of DC poisoning on the activity and expression of the superoxide dismutase （SOD） gene in rabbit testis and epi-didymis. [Method ] 48 male rabbits were randomly divided into Wank control group and experimental group I , II and III. The dried plant of 0. glabra DC was comminuted,then different amounts of the grass powder （15% ,30% and 45% and the corresponding swainsonine content was 30, 60, 90 mg/kg） were mixed with the forage in 3 experimental groups. The rabbits consumed the forages freely until typical symptoms were ob-served; the test period was 70 d. 4 rabbits were selected randomly from each group on the 14 th, 35 th and 70 th day respectively; the testis and epididymis were collected. The activity of SOD was measured by hydroxyla- mine method,the mRNA expression was measured by real - time quantitative polymerase chain reaction （ real -time PCR） , the level of SOD protein expression was measured by immunohistochemical staining and West-ern Blot. [Result ]The activity of SOD was measured by hydroxylamine method, the mRNA expression was measured by real - time quantitative polymerase chain reaction （ real - time PCR）,the level of SOD protein expression was measured by immunohistochemical staining and Western Blot. The result showed that the activ-ity of SOD, the expression of SOD mRNA and protein decreased in poisoning rabbits testis and epididymis were very significant lower than that of the control group （P 〈0.01） form 35 th day.[Conclusion] The results showed that 0. glabra DC has remarkable effects on the expression of SOD mRNA and protein in rabbit testis and epididymis, presenting a certain time dose effect. The activity and expression change in SOD is highly as-sociated with 0. glabra DC poisoning and it may play an important role in this process.
出处
《新疆农业科学》
CAS
CSCD
北大核心
2017年第8期1540-1549,共10页
Xinjiang Agricultural Sciences
基金
国家自然科学基金项目"小花棘豆中毒对和田羊抗氧化机能的影响机制"(31460678)~~
关键词
小花棘豆
家兔
睾丸
附睾
超氧化物歧化酶
short ultivars
growth stages
major gene plus polygene inheritance
genetic analysis
