PRMT1对哮喘小鼠的影响以及作用机制分析

Explore PRMT1' influence in asthma mice and related mechanism

目的探讨蛋白质精氨酸甲基转移酶1(PRMT1)对哮喘的影响及作用机制。方法构建小鼠哮喘模型,然后分别利用氨茶碱(AMI)和磷酸盐缓冲液(PBS)处理两组小鼠,收集并用ELISA试剂盒检测小鼠支气管肺泡灌洗液中炎症因子白细胞介素(IL)-1β、IL-4和肿瘤坏死因子α(TNF-α)的水平。运用Bradford方法检测小鼠支气管肺泡灌洗液中蛋白含量。Western blotting检测肺脏组织中PRMT1的含量以及NF-κB信号通路下游分子P65的变化。结果哮喘模型组IL-4,IL-1β和TNF-α的水平较对照组明显升高(25.48±4.96 pg/ml vs.4.01±0.56 pg/ml,23.48±6.11 pg/ml vs.3.77±0.27 pg/ml,29.45±6.98 pg/ml vs.5±1.84 pg/ml);经AMI处理的哮喘模型组IL-4,IL-1β,TNF-α的水平较PBS处理的哮喘模型组明显降低(12.14±3.87 pg/ml vs.25.48±4.96 pg/ml,11.95±2.55 pg/ml vs.23.48±6.11 pg/ml,16.81±7.49 pg/ml vs.29.45±6.98 pg/ml),差异有统计学意义(P<0.05)。哮喘模型组小鼠支气管肺泡灌洗液中蛋白渗出量较对照组明显增加,差异有统计学意义(P<0.05),同时对比PBS处理组和AMI处理组发现,AMI处理后小鼠支气管肺泡灌洗液中蛋白渗出含量明显减少(P<0.05)。Western blotting检测了四组小鼠肺脏组织中PRMT1和P65的蛋白含量,结果显示在患有哮喘的小鼠中PRMT1和P65的表达水平较对照组显著升高,无论是对照组还是哮喘模型组,AMI处理后的PRMT1和P65的表达水平较PBS处理组降低。结论 PRMT1通过上调NF-κB信号通路促进肺脏炎性因子IL-1β、IL-1和TNF-α的产生以及蛋白渗出,诱导哮喘的发生。

Objective To investigate the protein arginine methyl transferase 1（ PRMT1） effect on Asthma and the its action mechanism. Methods Mice asthma disease model was constructed,and then using the AMI and PBS in two groups of mice respectively,collect bronchoalveolar lavage fluid to detect the level of IL- 4,IL- 1 β,TNF- α with ELISA kits. Bradford was used to detect protein content in bronchoalveolar lavage fluid in mice. Western blotting was used to determined PRMT1 and P65 content in lung tissue. Results The level of IL- 4,IL- 1 β and TNF- α in mice asthma group were higher than those of the control group（ 25. 48 ± 4. 96 pg / ml vs. 4. 01 ± 0. 56 pg / ml,23. 48 ± 6. 11 pg / ml vs. 3. 77 ± 0. 27 pg / ml,29. 45 ± 6. 98 pg / ml vs. 5 ± 1. 84 pg / ml）. The level of IL- 4,IL- 1 β and TNF- α in asthma model group with AMI treatment were lower than PBS treatment group（ 12. 14 ± 3. 87 pg / ml vs. 25. 48 ± 4. 96 pg / ml,11. 95 ± 2. 55 pg / ml vs. 23. 48 ± 6. 11 pg / ml,16. 81 ± 7. 49 pg / ml vs. 7. 49 ± 6. 98 pg / ml）,the difference was statistically significant（P〈0. 05）. The protein in bronchoalveolar lavage fluid seepage in asthma model group mice increased significantly than that in the control group,the difference was statistically significant（P〈0. 05）,and compared with the PBS treatment group,AMI treatment effusion protein content in bronchoalveolar lavage fluid in mice decreased significantly（P〈0. 05）. Western blotting examined the four groups of PRMT1 and P65 protein content in the lung tissue of mice,the results showed that the expression level of PRMT1 and P65 protein in mice suffering asthma significantly increased than that in the control group,PRMT1 and P65 expression level was lower after AMI treatment than those of PBS treatment group. Conclusion PRMT1 can promote lung inflammatory factor and the production of IL- 4,IL- 1 β,TNF- α and effusion protein level by actinvating the NF- ΚB signaling pathway,which is an important target of lung epithelial inflammation.