厄洛替尼对脂多糖诱导的巨噬细胞炎症反应和小鼠急性肺损伤的影响

目的: 探讨厄洛替尼对脂多糖(LPS)诱导巨噬细胞产生的炎症反应和小鼠急性肺损伤(ALI)的影响,阐明其作用机制。方法: 原代培养的小鼠骨髓来源的巨噬细胞随机分为对照组、厄洛替尼组、LPS组和LPS+厄洛替尼组。采用酶联免疫吸附(ELISA)法检测各组巨噬细胞中肿瘤坏死因子α(TNF-α)水平;Western blotting法检测各组巨噬细胞中ERK1/2和p38磷酸化水平。16只C57BL/6小鼠随机分为对照组(生理盐水灌胃3d,腹腔注射生理盐水1次)、厄洛替尼组(45mg·kg^-1厄洛替尼预处理3d,腹腔注射生理盐水1次)、LPS组(生理盐水灌胃3d,腹腔注射5mg·kg^-1LPS)和LPS+厄洛替尼组(45mg·kg^-1厄洛替尼连续3d灌胃,腹腔注射5mg·kg^-1LPS)。ELISA法检测各组小鼠血清中TNF-α的表达水平;Western blotting法检测各组小鼠肺组织中ERK1/2和p38磷酸化水平;HE染色观察各组小鼠肺组织病理形态表现。结果: 与对照组比较,厄洛替尼组巨噬细胞和小鼠血清中TNF-α水平、ERK1/2和p38蛋白磷酸化水平差异无统计学义(P>0.05),小鼠肺组织形态无明显改变;与厄洛替尼组比较,LPS组巨噬细胞和小鼠血清中TNF-α水平明显升高(P<0.05),巨噬细胞和肺组织中ERK1/2和p38蛋白磷酸化水平明显升高(P<0.05),小鼠肺组织出现ALI改变;与LPS组比较,LPS+厄洛替尼组巨噬细胞和小鼠血清中TNF-α表达水平明显降低(P<0.05),巨噬细胞和肺组织中ERK1/2和p38蛋白磷酸化水平降低(P<0.05),小鼠肺组织ALI病理状态改善。结论: 厄洛替尼可以抑制巨噬细胞炎症通路蛋白ERK1/2和p38的磷酸化水平及炎症因子TNF-α的生成,降低ALI小鼠的全身炎症反应,在一定程度上对ALI有保护作用。

Effects of erlotinib on inflammatory response of marcrophages and ALI induced by lipopolysaccharides in mice

Objective: To investigate the effects of erlotinib on the inflammatory response of macrophages and acute lung injury(ALI ) induced by lipopolysaccharides(LPS)in the mice,and to reveal their mechanisms. Methods: The murine bone marrow derived macrophages were randomly divided into control group,erlotinib group,LPS group, and LPS+erlotinib group. The expression levels of tumor necrosis factor α(TNF-α) in the macrophages in various groups were detected by ELISA method and the phosphorylation levels of p38 and ERK1/2 were tested by Western blotting method. A total of 16 male mice were randomly divided into control group(saline),erlotinib group(45 mg·kg^-1 erlotinib for 3 d+saline),LPS group(saline+5 mg·kg^-1 LPS),LPS+erlotinib group(45 mg·kg^-1erlotinib+5 mg·kg^-1 LPS). The expression levels of TNF-α in the macrophages in serum was detected by ELISA method and the phosphorylation levels of p38 and ERK1/2 were tested by Western blotting method;the pathomorphology of lung tissue of the mice in various groups was observed by HE staining. Results: Compared with control group, the expression levels of TNF-α in the macrophages and serum of the mice,and the phosphorylation levels of ERK1/2 and p38 in the macrophages and lung tissue of the mice in erlotinib group had no significant difference(P>0.05),and the pathomorphology of lung tissue of the mice had no changes.Compared with erlotinib group,the expression levels of TNF-α in the macrophoages and serum of the mice and the phosphorylation levels of ERK1/2 and p38 in the macrophages and lung tissue of mice in LPS group were significantly increased(P<0.05),and the pathomorphology of LPS-induced ALI could be found.Compared with LPS group,the expression levels of TNF-α in the macrophages and serum of the mice and the phosphorylation levels of ERK1/2 and p38 in the macrophages and lung tissue of the mice in LPS+erlotinib group were decreased(P<0.05),and the pathomorphology of ALI was improved. Conclusion: Erlotinib can reduce the systemic inflammatory response of the ALI mice by inhibiting the phosphorylation levels of p38 and ERK1/2 of macrophages and the expression level of TNF-α to protect the ALI to a certain degree.