HSF1对LPS诱导的急性肺损伤小鼠的保护作用及其相关差异表达基因的筛选

摘要

AIM:To study the protective effect of heat shock factor1(HSF1) on the mice with lipopolysaccha-ride (LPS)-induced acute lung injury(ALI),and to screen the relevant differentially-expressed genes. METHODS:ALI mouse model was established by LPS intracheal instillation. The macroscopic and pathological changes of the lung tissue were observed,and the concentrations of total protein,TNF-α, IL-β, IL-6 and VEGF in the bronchoalveolar lavage fluid (BALF) were analyzed. Differentially-expressed genes in the lung tissues of HSF1 +/ +mice and HSF1 -/- mice with ALI induced by LPS were screened by gene chips. The key gene was verified by real-time qPCR. RESULTS:The macroscopic and pathological changes of the lung injury in HSF1 -/- +LPS mice were more serious than those in HSF1 +/ ++LPS mice.The concentrations of total protein,VEGF,TNF-α,IL-1β and IL-6 in the BALF of HSF1 -/- +LPS mice were significantly higher than those of HSF1 +/ ++LPS mice(P<0.05). Compared with the HSF1 +/ +mice,a total of 918 differentially-ex-pressed genes were indentified in the HSF1 -/- mice, among which the expression levels of 65 genes had obvious diffe-rence,with 28 genes up-regulated,including Atg7,ccr1,cxcr2,Tbl1xr1,Mmp9,Pparg,Plcb2,Arrb2,Cntn1,Col4a6, etc, and 37 genes down-regulated,including Fgfr1,Fgfr2,Map4k4,Ddx58,Tfg,Stat3,Smad4,Lamc1,Sdc3,etc. The results of real-time qPCR showed that the mRNA level of CXCR2 in HSF1 -/- + LPS mice was significantly higher than that in HSF1 +/ ++ LPS mice,which was consistent with the results of gene chips. CONCLUSION:HSF1 has protective effect on the mice with LPS-induced ALI. CXCR2 may be involved in the protective effect of HSF1 on this process.%目的:探讨热休克因子1(heat shock factor 1,HSF1)减轻脂多糖(lipopolysaccharide,LPS)诱导的小鼠急性肺损伤的作用及其分子机制.方法:采用气管滴注LPS的方法制备小鼠急性肺损伤模型,观察HSF1野生型小鼠(HSF1+/+)和HSF1敲除小鼠(HSF1-/-)肺大体改变和肺组织病理改变,检测支气管肺泡灌洗液(bronchoalve-olar lavage fluid,BALF)中总蛋白、血管内皮生长因子(vascular endothelial growth factor,VEGF)、肿瘤坏死因子 α(tumor necrosis factor-α,TNF-α)、白细胞介素(interleukin,IL)-1β和IL-6的蛋白表达.采用基因芯片技术筛选经LPS处理后的HSF1+/+和HSF1-/-小鼠肺组织中的差异表达基因,并进一步采用real-time PCR对CXC趋化因子受体2(CXC chemokine receptor 2,CXCR2)的表达进行验证.结果:与经LPS刺激后的HSF1+/+小鼠相比,经LPS刺激的HSF1-/-小鼠肺大体和病理损伤加重,BALF中总蛋白、VEGF、TNF-α、IL-1β和IL-6的含量升高,差异具有统计学意义(P<0.05).基因芯片分析发现,与经LPS处理的HSF1+/+小鼠相比,HSF1-/-小鼠共筛选出918个差异基因,有65个基因表达差异明显,其中Atg7、ccr1、cxcr2、Tbl1xr1、Mmp9、Pparg、Plcb2、Arrb2、Cntn1、Col4a6等共28个基因在HSF1-/-小鼠的肺组织中表达明显上调;Fgfr1、Fgfr2、Map4k4、Ddx58、Tfg、Stat3、Smad4、Lamc1、Sdc3等共37个基因表达明显下调.Real-time PCR 结果显示,CXCR2的 mRNA 水平在 LPS 刺激的hSF1-/- 小鼠肺组织较HSF1+/+小鼠表达明显上调,表达趋势与基因芯片结果一致.结论:HSF1能减轻LPS诱导的小鼠急性肺损伤,CX-CR2可能参与了对肺组织的保护作用.