摘要
目的研究紫花牡荆素对脓毒症小鼠急性肺损伤(ALI)的影响,并探讨其机制。方法将60只C57BL/6小鼠随机分为假手术组、模型组、紫花牡荆素低(2.5 mg/kg)、中(5 mg/kg)、高(10 mg/kg)剂量组,每组12只。除假手术组外,其余各组小鼠均采用盲肠结扎穿刺(CLP)法建立脓毒症模型,模型构建成功后连续给药2 d。采用血气分析仪检测小鼠动脉血氧分压(PaO_(2))和动脉血CO_(2)分压(PaCO_(2));分离小鼠右肺下叶,测定湿/干重比值;苏木精-伊红(HE)染色观察肺组织病理变化;ELISA法测定肺泡灌洗液(BALF)中肿瘤坏死因子-α(TNF-α)、白细胞介素1β(IL-1β)和IL-6水平;Fluo-4/AM荧光探针标记法检测肺组织细胞内Ca^(2+)浓度;Western blot检测肺组织中钙通道调节蛋白1(Orai1)、CCAAT/增强子结合蛋白同源蛋白(CHOP)和葡萄糖调节蛋白78(GRP78)的蛋白表达水平。结果与假手术组比较,模型组小鼠PaO_(2)显著降低(P<0.05),而PaCO_(2)和肺组织湿/干重比值显著升高(均P<0.05),肺组织结构紊乱,肺泡壁增厚,肺泡塌陷,肺间质水肿,有炎性细胞浸润,BALF中TNF-α、IL-1β和IL-6水平均显著升高(均P<0.05),肺组织细胞内Ca^(2+)浓度及肺组织中Orai1、CHOP和GRP78蛋白表达水平均显著升高(均P<0.05)。与模型组比较,紫花牡荆素各剂量组小鼠PaO_(2)升高(P<0.05),PaCO_(2)和肺组织湿/干重比值显著降低(均P<0.05),肺组织病理损伤减轻,BALF中TNF-α、IL-1β和IL-6水平显著降低(均P<0.05),肺组织细胞内Ca^(2+)浓度及肺组织中Orai1、CHOP和GRP78蛋白表达水平也显著降低(均P<0.05)。并且,高剂量紫花牡荆素对各指标的作用比低剂量更显著(均P<0.05)。结论紫花牡荆素可改善脓毒症小鼠ALI,抑制炎症反应,其作用机制可能与抑制Ca^(2+)内流,减轻内质网应激有关。
Objective To investigate the effect of casticin on acute lung injury(ALID)in sepsis mice,and explore its mecha-nism.Methods Sixty C57BL/6 mice were randomly divided into sham operation group,model group,low(2.5 mg/kg),medium(5 mg/kg),and high-dose(10 mg/kg)of casticin groups,with 12 mice in each group.Except for the sham operation group,the mice in the other groups were used to establish sepsis models using cecal ligation and puncture(CLP)method.After successful model construction,the mice were continuously administered for 2 days.Blood gas analyzer was used to detect arterial partial pressure of oxygen(PaO_(2))and arterial partial pressure of CO_(2)(PaCO_(2))in mice.The lower lobe of right lung was isolated and the ratio of wet to dry weight was measured.HE staining was used to observe the pathological changes in lung tissue.ELISA was used to measure the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1p),and IL-6 in alveolar lavage fluid(BALF).Fluo-4/AM fluorescence probe labeling method was used to detect intracellular Ca^(2+)concentration in lung tissue cells,Western blot was used to detect the protein expression levels of calcium channel regulatory protein 1(Orail),CCAAT/enhancer binding protein homologous protein(CHOP)and glucose regulatory protein 78(GRP78)in lung tissue.Results Compared with the sham surgery group,mice in model group showed a significant decrease in PaO,(P<0.05),while PaCO_(2)and lung tissue wet to dry weight ratio significantly increased(all P<0.05).The lung tissue structure was disrupted,with thickening of alveolar walls,col-lapse of alveoli,interstitial edema,and inflammatory cell infiltration.Levels of TNF-α,IL-1βand IL-6 in BALF were significantly increased in lung tissue(all P<0.05).The intracellular Ca^(2+)concentration in lung tissue cells and the protein expression levels of Orail,CHOP,and GRP78 in lung tissue were also significantly increased(all P<0.05).Compared with the model group,the PaO_(2)of mice in each dose of casticin group increased(all P<0.05),and PaCO_(2)and lung tissue wet to dry weight ratio decreased significantly(all P<0.05).The pathological damage to lung tissue was alleviated,and the levels of TNF-α,IL-1β and IL-6 in BALF were significantly reduced(all P<0.05).The intracellular Ca+concentration in lung tissue cells and the protein expres-sion levels of Orail,CHOP and GRP78 in lung tissue were also significantly reduced(all P<0.05).Effects of high-dose casticin on various indicators were more significant than that of low-dose casticin(all P<0.05).Conclusion Casticin can improve ALI and inhibit inflammatory response in sepsis mice,and its mechanism may be related to Ca+influx inhibition and endoplasmic re-ticulum stress alleviation.
作者
闫智杰
游燕
Yan Zhijie;You Yan(Department of Critical Care Medicine,Fourth Affiliated Hospital of Nanchang University,Nanchang 330003,China;Department of Pharmacy,Fourth Af filiated Hospital of Nanchang University,Nanchang 330003,China)
出处
《华中科技大学学报(医学版)》
北大核心
2025年第4期477-482,共6页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
江西省卫生健康委员会科技计划项目(No.SKJP220211237)。
