JNK信号通路在高氧诱导A549细胞结缔组织生长因子表达中的作用

Role of JNK signaling pathway in hyperoxia-induced expression of connective tissue growth factor in A549 cells

目的探讨c-Jun氨基末端激酶（JNK）信号通路在高氧诱导A549细胞结缔组织生长因子（CTGF）表达中的作用。方法培养的A549细胞以1×10^5个／ml密度接种于6孔培养板，0．5ml／孔，共24孔，采用随机数字表法分为4组（n=6）：空气对照组（c组）、95％氧气组（95％O2组）、空气＋JNK抑制剂SP600125组（C＋SP组）和95％氧气＋SP600125组（95％O2＋SP组）。C＋SP组和95％O2＋SP组每孔加入SP6001255μmol／L。将A549细胞分别置于空气（c组和C＋SP组）和95％氧气（95％O2组和95％O2＋SP组）中进行培养48h，采用Western blot法测定磷酸化JNK（p-JNK）的表达；采用RT-PCR法测定CTGFmRNA的表达。结果与C组比较，C＋SP组细胞p-JNK和CTGFmRNA表达差异无统计学意义（P〉0．05），95％O2组P—JNK和CTGFmRNA表达上调（P〈0．05）；与95％O2组比较，95％O2 ＋SP组p-JNK和CTGFmRNA表达下调（P〈0．05）。结论JNK信号通路激活参与了高氧诱导A549细胞CTGFmRNA表达上调的过程。

Objective To investigate the role of c-Jun N-terminal kinase （JNK） signaling pathway in hyperoxia-induced expression of connective tissue growth factor （CTGF） in A549 cells. Methods The cultured A549 ceils were seeded in 6-well culture plates at a density of 1 × 10^5 cells/ml （0.5 ml/well, 24 wells in total） and were randomly divided into 4 groups （n = 6 each） using a random number table： control group （group C） ; 95% oxygen group （group 95% 02） ; room air plus SP600125 （JNK inhibitor） group （group C＋SP） ; 95% oxygen plus SP600125 group （group 95% O2＋SP）. SP600125 5 μmol/L was added to each well in C＋SP and 95% O2＋SP groups. The A549 cells were incubated for 48 h in an incubator filled with room air （C and C＋SP groups） or 95% oxygen （95% 02 and 95% O2＋SP groups）. The expression of phosphorylated JNK （p-JNK） and CTGF mRNA was determined using Western blot and real- time reverse transcriptase polymerase chain reaction, respectively. Results Compared with group C, no significant change was found in the expression of p-JNK and CTGF mRNA in group C＋SP （P〉0.05） , and the expression of p-JNK and CTGF mRNA was significantly up-regulated in group 95% O2 （P〈0.05）. Compared with group 95% 02, the expression of p-JNK and CTGF mRNA was significantly down-regulated in group 95% O2＋SP （P〈0.05）. Conclusion JNK signaling pathway activation is involved in up-regulation of hyperoxia-induced CTGF mRNA expression in A549 ceils.