晚期糖基化终末产物受体-核因子κB 信号通路在脂多糖致新生大鼠急性肺损伤的作用

Role of receptor for advanced glycation end - products nuclear factor - κB signaling regulating pathway in li-popolysaccharide-induced acute lung injury in neonatal rats

目的：探讨晚期糖基化终末产物受体-核因子κB（RAGE-NF-κB）信号通路在脂多糖（LPS）致新生大鼠急性肺损伤（ALI）中的作用。方法32只 SD 新生大鼠完全随机化分为4组，每组8只。（1）LPS 组：腹腔注射9 g/ L 盐水0.2 mL，1 h 后腹腔注射3 mg/ kg LPS；（2）硼替佐米组：腹腔注射0.2 mg/ kg 硼替佐米，1 h 后腹腔注射3 mg/ kg LPS；（3）抗 RAGE 单抗组：腹腔注射15 mg/ kg 抗 RAGE 单抗，1 h 后腹腔注射3 mg/ kg LPS；（4）对照组：在相应的时间点腹腔注射相同容积的9 g/ L 盐水。观察大鼠一般情况，24 h 后麻醉后处死全部动物，酶联免疫吸附法检测血清及支气管肺泡灌洗液（BALF）中肿瘤坏死因子α（TNF-α）水平；Western blot 检测肺组织 RAGE 及 NF-κB 蛋白的表达；反转录-聚合酶链反应法检测肺组织 RAGE mRNA 及 NF-κB mRNA 的表达；HE 染色观察肺组织病理变化。结果1.血清及 BALF 中 TNF-α水平：4组 TNF-α水平总体差异存在统计学意义（F =150.70，P ﹤0.001；F =165.83，P ﹤0.001）；LPS 组血清及 BALF 中 TNF-α水平最高；与 LPS 组相比，2个预处理组 TNF-α水平均明显下降（P 均﹤0.05）。2.与 LPS 组相比，2个预处理组 RAGE、NF-κB 蛋白均明显下降。3. RAGE mRNA、NF-κB mRNA 表达：4组 RAGE mRNA、NF-κB mRNA 水平总体差异存在统计学意义（F =175.14， P ﹤0.05；F =188.65，P ﹤0.05）。与 LPS 组相比，2个预处理组 RAGE mRNA、NF-κB mRNA 均明显下降（P 均﹤0.05）。4.肺组织病理评分：4组病理评分总体差异存在统计学意义（F =106.01，P ﹤0.001）；LPS 组评分最高，损伤最严重；与 LPS 组相比，2个预处理组评分较低，差异均有统计学意义（P 均﹤0.05）。结论 RAGE-NF-κB 信号途径参与 LPS 诱导的 ALI，抗 RAGE 单抗、硼替佐米对 LPS 诱导的 ALI 有保护作用。

Objective To investigate the role of receptor for advanced glycation end - products nuclear factor - κB（RAGE - NF - κB）signaling pathway in the lipopolysaccharide - induced acute lung injury（ALI）in neo-natal rats. Methods Thirty - two SD rats were divided into 4 groups by complete randomization method（8 cases in each group）.（1）Lipopolysaccharide（LPS）group was given intraperitoneal injection of 9 g/ L saline and 3 mg/ kg LPS 1 h later.（2）Bortezomib group was given intraperitoneal injection of Bortezomib（0. 2 mg/ kg）and 3 mg/ kg LPS 1 h later.（3）Anti - RAGE mAb group was given intraperitoneal injection of anti - RAGE mAb（15 mg/ kg）and 3 mg/ kg LPS 1 h later.（4）Control group was given 9 g/ L saline was given at each time point. All the rats were sacrificed and observed 24 h later. Levels of tumor necrosis factor（TNF） - α in the plasma and bronchoalveolar lavage fluid（BALF） were detected by enzyme linked immunosorbent assay. RAGE and NF - κB levels in tissue homogenates were detected by Western blot and mRNA levels were detected by reverse transcription - polymerase chain reaction. The pathological assessment of the lung tissues was performed by HE staining. Results （1）Among 4 groups,there were significantly differences in TNF - α in serum and BALF（F = 150. 70,P ﹤ 0. 001;F = 165. 83,P ﹤ 0. 001）. Levels of TNF - α in LPS group were significantly higher than those of two pretreatment groups（all P ﹤ 0. 05）.（2）Western blot figures il-lustrated that the concentrations of RAGE mRNA and NF - κB in anti - RAGE mAb group and bortezomib group were lower than those of the LPS group.（3）Reverse transcription - polymerase chain reaction analysis showed that there were significant differences in the expression of RAGE mRNA and NF - κB mRNA among 4 groups（F = 175. 14,P ﹤0. 05;F = 188. 65,P ﹤ 0. 05）. Levels of RAGE mRNA and NF - κB mRNA in the LPS group were significantly higher than those of two pretreatment groups（all P ﹤ 0. 05）.（4）Lung injury score differences among 4 groups were statistical-ly significant（F = 106. 01,P ﹤ 0. 001）. Pathological changes in two pretreatment groups reduced compared to those of the LPS group（all P ﹤ 0. 05）. Conclusions RAGE - NF - κB signaling pathway regulates the LPS - induced ALI in neonatal rats. Anti - RAGE mAb and Bortezomib both have a protective effect on LPS - induced ALI.