免疫组织化学法定位猪脾脏中脱氧雪腐镰刀菌烯醇研究.docVIP

免疫组织化学法定位猪脾脏中脱氧雪腐镰刀菌烯醇研究 　　摘要 利用免疫组织化学法研究了脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)在猪脾脏分布定位,为DON体内残留及DON对免疫系统影响进一步研究提供依据。选用10 头健康阉公猪,随机分为二组,一组经乳静脉注射DON标准品,注射剂量为75 μg/kg体重,另一组经乳静脉注射相当体积生理盐水。注射完成后开始计时,并观察记录猪的反应情况,待30 min后放血宰杀,并迅速取脾脏,4%多聚甲醛固定,经脱水、包埋及切片后,采用免疫组化法定位脾脏中DON的分布情况。免疫组化法定位结果显示,在75 μg/kg体重给药组中,脾组织中的血管壁周围见有DON阳性反应颗粒,而对照组未见阳性反应颗粒。 　　关键词 脱氧雪腐镰刀菌烯醇;脾脏;猪;免疫组织化学法 　　中图分类号S852文献标识码A文章编号 1007-5739(2010)18-0292-02 　　 　　TheResearchofLocatingDONinSpleenofPigwithImmunohistochemical 　　LU Pei-chengHE Li-minNIU Geng-zhiYAN Hai-kuoMA Yong-jiangFAN Xiao-long LI Yu-guDENG Xian-bo * 　　(College of Veterinary Medicine,South China Agricultural University,Guangzhou Guangdong 510642) 　　AbstractThe distribution and location of deoxynivalenol in spleen of pigs by immunohistochemicalwasstudied, which provided a bisis for the residue of DON in the body and the effect of DON on the immune system. In the research,ten healthy clean pigs were used and divided into two groups randomly. One group were given DON standard by breast vein for the dose 75μg/kg b.w., normal saline was injected at the same dosage in the other group. Counted time after the intravenous administration, observed and noted the response,pigs were killed after 30 minutes, took spleen tissue sample quickly, fixed with 4% paraformaldehyde, dehydrated, embedded and sliced, then the immunohistochemical was used to locate the distribution of DON in spleen. The result of the study showed that DON positive particles were found in the spleen tissue around the blood vessel while in the control group no positive particles were saw. 　　Key wordsDON;spleen;pig;immunohistochemical 　　 　　脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON),又称作呕吐毒素(vomitoxin,VT),属单端孢霉烯族类,其中DON对谷物的污染率和污染水平居于镰刀菌毒素之首,人畜误食后会出现呕吐、腹泻、腹痛等症[1-2],也是中国恶性肿瘤高发区居民粮食优势污染霉菌毒素之一[3-4]。 　　随着研究的深入,发现所有的单端孢霉烯族毒素类毒素都有一个共同的特点:它们的免疫激活或者免疫抑制作用都有着剂量的依赖性,高剂量的单端孢霉烯族毒素类可以促进白细胞的快速凋亡,而这种凋亡则被称之为免疫抑制[5]。为进一步探讨DON在免疫系统的组织分布情况,该研究拟采用静注后脾脏取样、石蜡切片制作、免疫组织化学法[6]、显微镜观察等一系列步骤对脾脏中的DON的定位情况进行研究。 　　1材料与方法 　　1.1试验动物和切片制作