The cDNA of human Interleukin\|18 was obtained by RT\|PCR from the mRNA of human peripheral blood cells.The DNA sequencing was performed and the DNA was cloned into expression vector pBV220.The overexpression was perf...The cDNA of human Interleukin\|18 was obtained by RT\|PCR from the mRNA of human peripheral blood cells.The DNA sequencing was performed and the DNA was cloned into expression vector pBV220.The overexpression was performed in E.coli HB101.The expressive product was purified through Sephadex G100 and identified by N\|terminal amino acid sequencing and Western blot.展开更多
目的探讨白细胞介素18(interleukin 18,IL-18)对分泌性中耳炎(otitis media with effusion,OME)模型大鼠中耳炎症细胞的增殖情况以及干扰素γ(interferonγ,IFN-γ)和白细胞介素4(interleukin 4,IL-4)表达的影响。方法 18只SD大鼠随机...目的探讨白细胞介素18(interleukin 18,IL-18)对分泌性中耳炎(otitis media with effusion,OME)模型大鼠中耳炎症细胞的增殖情况以及干扰素γ(interferonγ,IFN-γ)和白细胞介素4(interleukin 4,IL-4)表达的影响。方法 18只SD大鼠随机分为对照组(A组)、OME模型组(B组)和IL-18注射组(C组),每组各6只(12耳),B组和C组采用卵清蛋白(ovalbumin,OVA)腹腔注射致敏后耳内激发制成OME模型,C组于OVA全身致敏及耳内激发的同时,在实验第1、2、7、8、15、16d进行腹腔注射IL-18 1μg加生理盐水0.2ml,A组和B组在相同时间点腹腔注射生理盐水0.2ml。HE染色观察各组中耳黏膜病理变化及炎症细胞的改变,ELISA法测定各组大鼠中耳腔灌洗液中T辅助细胞(T helper cell,Th)分泌Th2型细胞因子-IL-4和Th1型细胞因子-IFN-γ的含量。结果 C组大鼠中耳黏膜组织炎症反应较B组无明显减轻,中耳黏膜下层和骨髓腔中中性粒细胞数量较B组显著增多(P<0.05),嗜酸粒细胞数量与B组比较稍减少(P>0.05),但仍明显多于A组(P<0.05);C组中耳灌洗液Th1型细胞因子IFN-γ含量明显高于B组与A组(P<0.05),Th2型细胞因子IL-4含量较B组稍有增高(P>0.05);但B组和C组IL-4含量均明显高于A组(P<0.05);A、C组Th1/Th2比值虽高于B组,但差异无统计学意义(P>0.05)。结论 IL-18诱导OME模型大鼠中耳微环境中IFN-γ合成显著增高,虽然一定程度上缓解了OME大鼠中耳Th1/Th2失衡状态,但是仍存在以Th细胞过度活化、Th1和Th2型细胞因子过度分泌的免疫反应紊乱状态。展开更多
文摘The cDNA of human Interleukin\|18 was obtained by RT\|PCR from the mRNA of human peripheral blood cells.The DNA sequencing was performed and the DNA was cloned into expression vector pBV220.The overexpression was performed in E.coli HB101.The expressive product was purified through Sephadex G100 and identified by N\|terminal amino acid sequencing and Western blot.
