中华实用儿科临床杂志
中華實用兒科臨床雜誌
중화실용인과림상잡지
Journal of Applied Clinical Pediatrics
2015年
14期
1083-1086
,共4页
俞敏%唐云%赵赛%田兆方
俞敏%唐雲%趙賽%田兆方
유민%당운%조새%전조방
脂多糖%晚期糖基化终末产物受体%核因子-κB%急性肺损伤%大鼠,新生
脂多糖%晚期糖基化終末產物受體%覈因子-κB%急性肺損傷%大鼠,新生
지다당%만기당기화종말산물수체%핵인자-κB%급성폐손상%대서,신생
Lipopolysaccharide%Receptor for advanced glycation end - products%Nuclear factor - κB%Acute lung injury%Rat,newborn
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。
目的：探討晚期糖基化終末產物受體-覈因子κB（RAGE-NF-κB）信號通路在脂多糖（LPS）緻新生大鼠急性肺損傷（ALI）中的作用。方法32隻 SD 新生大鼠完全隨機化分為4組，每組8隻。（1）LPS 組：腹腔註射9 g/ L 鹽水0.2 mL，1 h 後腹腔註射3 mg/ kg LPS；（2）硼替佐米組：腹腔註射0.2 mg/ kg 硼替佐米，1 h 後腹腔註射3 mg/ kg LPS；（3）抗 RAGE 單抗組：腹腔註射15 mg/ kg 抗 RAGE 單抗，1 h 後腹腔註射3 mg/ kg LPS；（4）對照組：在相應的時間點腹腔註射相同容積的9 g/ L 鹽水。觀察大鼠一般情況，24 h 後痳醉後處死全部動物，酶聯免疫吸附法檢測血清及支氣管肺泡灌洗液（BALF）中腫瘤壞死因子α（TNF-α）水平；Western blot 檢測肺組織 RAGE 及 NF-κB 蛋白的錶達；反轉錄-聚閤酶鏈反應法檢測肺組織 RAGE mRNA 及 NF-κB mRNA 的錶達；HE 染色觀察肺組織病理變化。結果1.血清及 BALF 中 TNF-α水平：4組 TNF-α水平總體差異存在統計學意義（F =150.70，P ﹤0.001；F =165.83，P ﹤0.001）；LPS 組血清及 BALF 中 TNF-α水平最高；與 LPS 組相比，2箇預處理組 TNF-α水平均明顯下降（P 均﹤0.05）。2.與 LPS 組相比，2箇預處理組 RAGE、NF-κB 蛋白均明顯下降。3. RAGE mRNA、NF-κB mRNA 錶達：4組 RAGE mRNA、NF-κB mRNA 水平總體差異存在統計學意義（F =175.14， P ﹤0.05；F =188.65，P ﹤0.05）。與 LPS 組相比，2箇預處理組 RAGE mRNA、NF-κB mRNA 均明顯下降（P 均﹤0.05）。4.肺組織病理評分：4組病理評分總體差異存在統計學意義（F =106.01，P ﹤0.001）；LPS 組評分最高，損傷最嚴重；與 LPS 組相比，2箇預處理組評分較低，差異均有統計學意義（P 均﹤0.05）。結論 RAGE-NF-κB 信號途徑參與 LPS 誘導的 ALI，抗 RAGE 單抗、硼替佐米對 LPS 誘導的 ALI 有保護作用。
목적：탐토만기당기화종말산물수체-핵인자κB（RAGE-NF-κB）신호통로재지다당（LPS）치신생대서급성폐손상（ALI）중적작용。방법32지 SD 신생대서완전수궤화분위4조，매조8지。（1）LPS 조：복강주사9 g/ L 염수0.2 mL，1 h 후복강주사3 mg/ kg LPS；（2）붕체좌미조：복강주사0.2 mg/ kg 붕체좌미，1 h 후복강주사3 mg/ kg LPS；（3）항 RAGE 단항조：복강주사15 mg/ kg 항 RAGE 단항，1 h 후복강주사3 mg/ kg LPS；（4）대조조：재상응적시간점복강주사상동용적적9 g/ L 염수。관찰대서일반정황，24 h 후마취후처사전부동물，매련면역흡부법검측혈청급지기관폐포관세액（BALF）중종류배사인자α（TNF-α）수평；Western blot 검측폐조직 RAGE 급 NF-κB 단백적표체；반전록-취합매련반응법검측폐조직 RAGE mRNA 급 NF-κB mRNA 적표체；HE 염색관찰폐조직병리변화。결과1.혈청급 BALF 중 TNF-α수평：4조 TNF-α수평총체차이존재통계학의의（F =150.70，P ﹤0.001；F =165.83，P ﹤0.001）；LPS 조혈청급 BALF 중 TNF-α수평최고；여 LPS 조상비，2개예처리조 TNF-α수평균명현하강（P 균﹤0.05）。2.여 LPS 조상비，2개예처리조 RAGE、NF-κB 단백균명현하강。3. RAGE mRNA、NF-κB mRNA 표체：4조 RAGE mRNA、NF-κB mRNA 수평총체차이존재통계학의의（F =175.14， P ﹤0.05；F =188.65，P ﹤0.05）。여 LPS 조상비，2개예처리조 RAGE mRNA、NF-κB mRNA 균명현하강（P 균﹤0.05）。4.폐조직병리평분：4조병리평분총체차이존재통계학의의（F =106.01，P ﹤0.001）；LPS 조평분최고，손상최엄중；여 LPS 조상비，2개예처리조평분교저，차이균유통계학의의（P 균﹤0.05）。결론 RAGE-NF-κB 신호도경삼여 LPS 유도적 ALI，항 RAGE 단항、붕체좌미대 LPS 유도적 ALI 유보호작용。
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.
