CpG ODN通过活化p38 MAPK上调A549细胞β防御素2表达.docVIP

CpG ODN通过活化p38 MAPK上调A549细胞β防御素2表达 　 作者：李明，陈伟强，李岩，罗少洪，简玲敏 【摘要】 目的 研究含CpG基序的寡核苷酸(unmethylated CpG motif containing　oligodeoxynucleotide，CpG ODN)对人肺腺上皮细胞A549 β防御素2(human βdefensin 2,hBD2)的表达及p38 MAPK磷酸化水平的影响。方法 培养A549细胞，加或不加p38特异性抑制剂SB203580情况下用不同浓度CpG ODN进行干预。用RTPCR法检测刺激后A549细胞hBD2 mRNA表达变化，用Western Blot法检测细胞内p38 MAPK蛋白磷酸化水平的变化。结果 CpG ODN刺激可以促进A549细胞内磷酸化p38 MAPK含量的增加，活化p38 MAPK途径，从而诱导hBD2的表达，与对照组比较,Plt;0.05;用特异性阻断剂SB203580阻断p38 MAPK磷酸化可抑制CpG ODN的诱导作用。结论 CpG ODN通过p38 MAPK途径诱导hBD2在呼吸道上皮细胞的表达，增强呼吸道对外界微生物的抵抗作用。 【关键词】 CpG ODN; β防御素2;肺腺上皮细胞;p38 MAPK 论文代写 　Abstract:Objective To investigate the effects of unmethylated CpG motif containing　oligodeoxynucleotide (CpG ODN) on the expression of human βdefensin 2 (hBD2) and the phosphorylation of p38 MAPK in lung epithelial cell line A549. Methods A549 cells were stimulated with CpG ODN at different concentrations in the presence or absence of SB203580,an inhibitor of p38 MAPK. The mRNA expression of hBD2 was determined by RTPCR and the phosphorylation level of p38 MAPK was detected by Western Blot. Results CpG OND significantly enhanced the expression of hBD2 in A549 cells. The phosphorylation level of p38 MAPK was markedly upregulated. CpG ODNinduced hBD2 expression was suppressed by inhibition of p38 MAPK by SB203580. Conclusion CpG ODN might effectively upregulate hBD2 expression in lung epithelial cells through p38 MAPK pathway. 论文代写 　　Key words:CpG ODN;human βdefensin 2;lung epithelial cells;p38 MAPK 　　 目前认为，呼吸道上皮细胞不仅是物理性的防御屏障，还可以合成和分泌包括抗菌肽在内的多种细胞因子，在气道的炎症反应、免疫调节等方面发挥重要作用。β防御素2(human βdefensin 2,hBD2)是呼吸道上皮细胞在细菌和炎症因子诱导下所分泌的主要抗菌肽分子，不仅具有抗菌、抗病毒等生物学活性，还可以激活获得性免疫系统，在呼吸系统抵抗外界致病微生物感染中起重要作用[1]。如果能增强呼吸道hBD2的表达，对于防治呼吸系统的感染性疾病和应对日益严重的细菌耐药具有重要意义。细菌DNA与真核细胞不同，其含有大量的以非甲基化胞嘧啶鸟嘌呤二核苷酸(cytosine phosphateguanosine，CpG) 为核心的特定序列(即CpG基序)，因此可被真核细胞通过TLR9识别，最终激活NFκB、p38 MAPK等信号分子启动免疫应答[2,3]。目前人工合成的含CpG基序的寡核苷酸序列(unmethylated CpG motif containing　oligodeoxynucleotide，CpG ODN)已成为一种具有非特异性免疫刺激能力的免疫佐剂，能有效诱导T