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目的 探讨蛋白激酶C(PKC)和激活蛋白 -1(AP- 1)信号转导级联在支气管哮喘(简称哮喘)大鼠外周血T淋巴细胞白细胞介素5 (IL -5 )基因转录中的作用。方法 建立大鼠哮喘模型。从每只大鼠外周血中分离T淋巴细胞,并随机分为空白对照组、PKC激动剂12 肉豆蔻酰 13 乙酸佛波酯(PMA)组、PMA和AP -1顺式诱骗元件(CDODNs)组及PMA和AP -1突变顺式诱骗元件(突变CDODNs)组进行培养。两种CDODNs分别经脂质体转染至后两组细胞。用电泳迁移率改变分析(EMSA)检测AP -1活化,用逆转录聚合酶链反应(RT- PCR)检测IL -5mRNA表达。结果 (1)PMA组的AP -1活化(86 777.2 2±2 2 5 7.79)和IL 5mRNA表达(0 .810 0±0 .0 316 )增加,与空白对照组(分别为2 0 70 8.5 4±96 8.0 4和0 .30 5 0±0 .0 12 9)相比较,差异有统计学意义(P <0 .0 1)。(2 )PMA和AP -1顺式诱骗元件组的AP -1活化(2 3475 .90±75 7.99)被抑制,IL -5mRNA表达(0 .345 0±0 .0 12 9)亦减少,与PMA组相比较,其差异有统计学意义(P <0 .0 1)。PMA和AP 1突变顺式诱骗元件组的AP 1活化(86 95 5 .71±16 0 0 .38)和IL- 5mRNA表达(0 .8175±0 .0 2 2 2 )与PMA组相比较,差异无统计学意义(P >0 .0 5 )。(3)T淋巴细胞AP -1活化和IL 5mRNA表达呈显著正相关(P <0 .0 1)。结论 哮?
Objective To investigate the role of PKC and AP-1 signal transduction in the transcription of interleukin-5 (IL-5) gene in peripheral blood of bronchial asthmatic rats (asthma) effect. Methods The rat asthma model was established. T lymphocytes were isolated from peripheral blood of each rat and randomly divided into blank control group, PKC agonist 12 myristoyl acetate (PMA) group, PMA and AP-1 cotyledons (CDODNs) Group and PMA and AP-1 mutated cis-decoy elements (mutant CDODNs) group. Two CDODNs were transfected into the latter two groups by liposome respectively. AP-1 activation was detected by electrophoretic mobility shift assay (EMSA) and IL-5 mRNA expression was detected by reverse transcription polymerase chain reaction (RT-PCR). Results (1) AP-1 activation (86 777.2 2 ± 2 2 5 7.79) and IL 5 mRNA expression (0 8 010 ± 0 0 316) in PMA group increased compared with control group (20 70 8.5 4 ± 96 8.04 and 0.3050 ± 0.0129), the difference was statistically significant (P <0.01). (2) AP-1 activation (2 3475 .90 ± 75 7.99) in PMA and AP-1 cis-decoy components was also inhibited, and IL -5 mRNA expression (0.3450 ± 0.012 9) PMA group, the difference was statistically significant (P <0. 01). AP 1 activation (86 95 5 .71 ± 16 0 0 .38) and IL-5 mRNA expression (0 .875 ± 0 .0 2 2 2) in the PMA and AP 1 mutant cis-decoy element groups were significantly lower than those in the PMA group The difference was not statistically significant (P> 0.05). (3) The activation of AP-1 in T lymphocytes was positively correlated with the expression of IL-5 mRNA (P <0.01). Conclusion asthma?