目的 探讨姜黄素对胸部爆震致小鼠肺组织氧化应激损伤的影响。方法 将30只C57BL/6小鼠随机分为对照组、模型组、模型+姜黄素组,伤后24 h处死小鼠,左肺进行10%甲醛固定,石蜡包埋切片,右肺-80 ℃保存。HE染色观察肺组织爆震后病理改变;活性氧生成(reactive oxygen species,ROS)检测肺组织氧化应激损伤情况;Western blot检测氧化应激相关蛋白MDA5、SOD-1、IRE-α和信号通路相关蛋白p-PI3K、Nrf2、Keap1的表达。结果 胸部爆震后,模型组肺组织细胞排列明显紊乱、可见细胞变性和炎性细胞浸润;ROS检测显示,在紫外光激发下,模型组与对照组相比肺组织内蓝色荧光减少,红色荧光明显增加;Western blot检测结果显示模型组MDA5、IRE-α、SOD-1、Nrf2和Keap1的灰度值的相对表达分别为0.68±0.08、0.63±0.04、0.38±0.02、0.32±0.03和0.98±0.11,相比于对照组的0.33±0.03、0.29±0.02、0.81±0.09、0.43±0.05和0.44±0.04变化明显,而模型组p-PI3K/PI3K的灰度值的相对表达为0.74±0.05,相比于对照组的0.30±0.02显著升高,上述差异均有统计学意义(P<0.05)。姜黄素干预后,HE染色显示小鼠肺组织变性坏死明显缓解,炎性细胞浸润减轻;ROS检测显示模型+姜黄素组与模型组相比,肺组织内蓝色荧光增加,红色荧光明显减少;Western blot检测结果显示模型+姜黄素组MDA5、IRE-α、SOD-1、Nrf2和Keap1的灰度值的相对表达分别为0.48±0.07、0.36±0.06、0.51±0.07、0.71±0.08和0.53±0.06,p-PI3K/PI3K的灰度值的相对表达为0.56±0.07,上述差异与模型组两两比较均具有统计学意义(P<0.05)。结论 姜黄素对胸部爆震所致的肺组织氧化应激损伤有保护作用,其机制可能是通过PI3K/Nrf2/Keap1信号通路来实现的。
Abstract
Objective To investigate the effect of curcumin on oxidative stress in lung induced by thoracic blast exposure and its possible molecular mechanism.Methods Thirty C57BL/6 mice were randomly divided into control group,model group and model with curcumin group.Collect lung tissue at 24 h after thoracic blast exposure.The left lung was fixed in 10% formalin and embedded in paraffin.The right lung was preserved at -80 ℃.Hematoxylin-eosin staining was used to detect the pathological changes of lung tissue,and ROS was used to detect the oxidative stress injury.Western blot was used to detect the expression of MDA5,SOD-1,IRE-α,p-PI3K,Nrf2 and Keap1.Results After chest blast,the cell arrangement of lung tissue in model group was obviously disordered,cell degeneration and inflammatory cell infiltration were observed.Compared with the control group,the blue fluorescence decreased and the red fluorescence increased significantly in the model group under UV excitation.Western blot showed that the relative density of MDA5,IRE-α,SOD-1,Nrf2 and Keap1 in the model group were 0.68±0.08,0.63±0.04,0.38±0.02,0.32±0.03 and 0.98±0.11,respectively.These proteins in the control group were 0.33±0.03,0.29±0.02,0.81±0.09,0.43±0.05 and 0.44±0.04,respectively.The relative density of p- PI3K/PI3K in the model group was 0.74±0.05,which was significantly higher than that of the control group (0.30±0.02).The differences above were statistically significant (P<0.05).After curcumin intervention,comparing with the model group,HE staining showed that the degeneration and necrosis of lung tissue and the infiltration of inflammatory cells were alleviated.ROS detection showed that the blue fluorescence increased and the red fluorescence decreased significantly in the model with curcumin group.Western blot showed that the relative density of MDA5,IRE-α,SOD-1,Nrf2 and Keap1 in model with curcumin group were 0.48±0.07,0.36±0.06,0.51±0.07,0.71±0.08 and 0.53±0.06,respectively,and the relative density of p-PI3K/PI3K was 0.56±0.07.The above differences were statistically significant compared with the model group (P<0.05).Conclusions Curcumin can protect lung tissue from oxidative stress injury induced by thoracic blast exposure,and the mechanism may be associated with PI3K/Nrf2/Keap1 signaling pathway.
关键词
胸部爆震 /
姜黄素 /
肺损伤 /
氧化应激 /
PI3K/Nrf2/Keap1信号通路
Key words
thoracic blast exposure /
curcumin /
acute lung injury /
oxidative stress /
PI3K/Nrf2/Keap1 signaling pathway
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基金
2014总后卫生部重大新上(AWS14L008)
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