苯甲基磺酰氟对三邻甲苯磷酸酯暴露母鸡脑组织基因表达谱的影响

Effect of PMSF on gene expression profiles in the brain tissue of TOCP-treated hens

关键词:  有机磷诱发的迟发性神经毒性  苯甲基磺酰氟  三邻甲苯磷酸酯  脑  基因表达谱

Key words:  organophosphorus-induced delayed neurotoxicity (OPIDN)  phenylmethylsulfonyl fluoride (PMSF)  tri-ortho-cresyl phosphate (TOCP)  brain  gene expression profile

摘要:

［目的］从三邻甲苯磷酸酯（TOCP）暴露母鸡的脑组织中筛选迟发性神经毒性（OPIDN）相关基因，为探讨OPIDN发病机制提供靶基因信息谱。［方法］成年母鸡12只，随机分为1000 mg/kg TOCP暴露组、皮下注射40 mg/kg 苯甲基磺酰氟（PMSF）后再给TOCP的PMSF干预组及给生理盐水的对照组。染毒第5天处死鸡，取脑称重后制成匀浆，并依次行总RNA提取、纯化、体外转录cRNA探针合成、生物素标记、基因芯片杂交、杂交信号扫描和芯片数据处理分析。［结果］TOCP暴露组与对照组比较差异有显著性意义 （P<0.05），且与PMSF前干预组比较差异无显著性意义（P>0.05）的基因有311个。其中，差异表达3倍以上的36个。［结论］通过PMSF干预，从TOCP暴露母鸡脑组织中筛出与OPIDN诱发相关基因311个。其中，有36个基因可能与OPIDN密切相关。

Abstract:

［Objective］To screen the organophosphorus-induced delayed neurotoxicity (OPIDN)-related genes from the brain tissue of tri-ortho-cresyl phosphate (TOCP)-treated hens by means of pretreatment of phenylmethylsulfonyl fluoride (PMSF) and to provide the information of target gene profiles for exploring mechanisms of OPIDN.［Methods］Twelve hens were randomly divided into three groups including TOCP group,PMSF interfering group and control group.Hens in the TOCP group were treated with 1000 mg/kg TOCP.Hens in the PMSF interfering group were given with TOCP after 40 mg / kg PMSF administration.The hensin control group were received the saline.On the 5th day,the hens were sacrificed.The brains were dissected and homogenized.Total RNA in the brain tissue was extracted, purifled,and transcribed to cRNA probe.Then cRNA probe was biotin labeled and hybridized with Chicken Genome 430 2.0 Araay.Hybridization signals were scaned and genechip data was analyzed.［Results］It was found that 311 genes were significant difference compared with control group,but no significant difference compared with PMSF interfering group.There were 36 genes over 3 fold expressions among these genes found.［Conclusion］The 311 genes may be related to OPIDN in the brain tissue of TOCP-treated hens.Among these genes,the 36 genes may be closely associated with OPIDN.