富马酸氯马斯汀对肠缺血再灌注小鼠急性肺损伤的保护作用及机制研究

Protective effect and mechanism of clemastine fumarate on acute lung injury in mice with intestinal ischemia⁃reperfusion

目的:评价富马酸氯马斯汀(CLE)对肠缺血再灌注(I/R)小鼠急性肺损伤(ALI)的保护作用及其机制。方法:将24只SPF级Balb/c小鼠随机分为假手术组(Sham组)、缺血再灌注组(I/R组)、富马酸氯马斯汀预处理组(I/R+C组),I/R组建立肠缺血再灌注模型(缺血40 min,再灌注2 h),I/R+C组手术开始前腹腔注射CLE 5 mg/kg。采用HE染色观察肺组织形态并评分,测定肺湿重/干重(W/D)比值,ELISA法测定各组肺组织丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-px)、核因子-κB(NF-κB)及肿瘤坏死因子(TNF-α)水平,Western blot法测定肺内TLR4的表达量。结果:与Sham组相比,I/R组肺损伤评分及W/D比值增加,肺内MDA水平增加,SOD、GSH-px水平降低,NF-κB、TNF-α水平增加,TLR4表达上调,差异均具有统计学意义(P<0.05);与I/R组相比,I/R+C组肺损伤评分及W/D比值降低,肺内MDA水平降低,SOD、GSH-px水平增加,NF-κB、TNF-α水平降低,TLR4的表达下调,差异均具有统计学意义(P<0.05)。结论:富马酸氯马斯汀可减轻小鼠肠缺血再灌注后急性肺损伤,其机制可能与抑制肺组织氧化应激和炎症反应有关。

Objective:To evaluate the protective effect and mechanism of clemastine fumarate(CLE)on acute lung injury(ALI)in intestinal ischemia‐reperfusion(I/R)mice.Methods:Twenty‐four SPF Balb/c mice were randomly divided into sham operation group(sham group),ischemia‐reperfusion group(I/R group),and clemastine fumarate pretreatment group(I/R+C group).In the I/R group,an intestinal ischemia‐reperfusion model was established(ischemia for 40 minutes,reperfusion for 2 hours).In the I/R+C group,CLE 5 mg/kg was intraperitoneally injected before the operation.Lung tissue morphology was ob‐served and scored by HE staining;and the ratios of wet weight to dry weight(W/D)were recorded.the levels of MDA,SOD,GSH‐px,NF‐κB and TNF‐αin lung tissue of each group were determined by ELISA;Western blot method was used to deter‐mine the expression of TLR4 protein in lung tissue.Results:Compared with the Sham group,the I/R group had significantly high‐er lung tissue injury score and wet/dry ratio(P<0.05),increased lung tissue MDA level(P<0.05),decreased SOD and GSH px levels(P<0.05),and increased NF‐κB and TNF‐αlevels,the expression of TLR4 protein in lung tissue increased(P<0.05);compared with the I/R group,the lung tissue injury score and wet/dry ratio of the I/R+C group decreased(P<0.05),the level of MDA in lung tissue decreased(P<0.05),the levels of SOD and GSH‐px increased(P<0.05),and the levels of NFκB and TNF‐αdecreased(P<0.05),the expression of TLR4 protein in lung tissue decreased(P<0.05).Conclusion:Clemastine fumarate can alleviate acute lung injury after intestinal ischemia‐reperfusion in mice,and the mechanism may be related to the inhi‐bition of oxidative stress and inflammatory response in lung tissue.