摘要
背景烟雾吸入致肺纤维化是火灾后常见并发症,病死率高,目前缺乏有效的治疗手段,探索该重症的治疗药物及其机制迫在眉睫。目的通过构建烟雾吸入性急性肺损伤(smoke inhalation induced acute lung injury,SI-ALI)后肺纤维化大鼠模型,探讨羟氯喹(hydroxychloroquine,HCQ)是否通过抑制Wnt/β-catenin信号通路发挥抗肺纤维化作用。方法将140只雄性SD大鼠随机分为正常对照组(C组)、模型组(S组)、磷酸缓冲盐溶液组(S+PBS组)、糖皮质激素组(S+MP组)和羟氯喹组(S+HCQ组),其中S+HCQ组根据HCQ剂量不同(5 mg/kg、10 mg/kg、20 mg/kg)分为3个亚组,每组20只大鼠。各组大鼠每日实验前1 h,腹腔注射对应剂量的PBS、MP和HCQ,后置于实验箱中开始吸入烟雾,持续30 min,共吸入7 d,继续正常饲养至28 d,比较各组大鼠的存活率、肺组织病理变化;分别用qRT-PCR和Western blot检测大鼠肺组织中α-肌动蛋白(smooth muscle actin,α-SMA)、Ⅰ/Ⅲ型胶原纤维(collagenⅠ/Ⅲ,ColⅠ/Ⅲ)、β-catenin、糖原合酶激酶3β(p-GSK-3β/GSK-3β)的mRNA和蛋白水平。结果与C组(100%)相比,S组和S+PBS组大鼠吸入烟雾后存活率显著下降,72 h后存活率维持在70%(P<0.05),且病理提示大鼠肺纤维化程度升高。与S+PBS组相比,S+MP组和S+HCQ组大鼠存活率有所提高同时伴有肺纤维化程度减轻,其中20 mg/kg HCQ及4 mg/kg MP对大鼠存活率影响较大(分别为90%和95%),但只有后者差异有统计学意义(P<0.05)。Western blot结果提示,HCQ和MP干预后,大鼠肺组织中纤维化指标CollagenⅠ、CollagenⅢ、α-SMA和Wnt/β-catenin信号通路中核心蛋白指标β-catenin、p-GSK-3β/GSK-3β表达降低(P<0.05),其中20 mg/kg HCQ对蛋白的抑制作用最显著(P<0.05)。qRT-PCR实验结果与Western blot基本一致。结论HCQ对烟雾吸入致肺纤维化大鼠具有保护作用,且20 mg/kg HCQ的保护效果最好,其中部分保护机制可能是通过抑制Wnt/β-catenin信号通路实现。
Background Pulmonary fibrosis due to smoke inhalation is a common complication after fire,with high mortality rate and a lack of effective treatments,making it urgent to explore therapeutic agents and their mechanisms for this serious condition.Objective To investigate whether HCQ exerts anti-pulmonary fibrosis effect through inhibiting Wnt/β-catenin signaling pathway by constructing a rat model of pulmonary fibrosis following smoke inhalation induced acute lung injury(SI-ALI).Methods Totally 140 male SD rats were randomly divided into control group(C),model group(S),phosphate buffered saline group(S+PBS),methylprednisolone group(S+MP)and hydroxychloroquine group(S+HCQ),among which the S+HCQ group was divided into 3 subgroups according to different HCQ doses(5 mg/kg,10 mg/kg,20 mg/kg),with 20 rats in each group.Rats in each group were given intraperitoneal injections of corresponding doses of PBS,MP,and HCQ at 1 h before smoke inhalation in the chamber daily for 30 min and a total of 7 d,and continued to be housed normally until 28 d for comparison of survival rates and lung histopathological changes among rats in each group;The mRNA and protein levels ofα-SMA,ColⅠ/Ⅲ,β-catenin and p-GSK-3β/GSK-3βwere detected in rat lung tissues by qRT-PCR and western blotting.Results Compared with the C group,the survival rate of rats in the S group and S+PBS group decreased significantly after smoke inhalation,and the survival rate remained at 70%after 72 h(P<0.05),and the pathology suggested an increase in pulmonary fibrosis.Compared with the S+PBS group,the survival rate of rats in the S+MP group and S+HCQ group increased and the degree of pulmonary fibrosis decreased,with 20 mg/kg HCQ and 4 mg/kg MP having obvious effects on the survival rate(90%and 95%),but only the 4 mg/kg MP group showed significant difference.Western blot results indicated that the expression of CollagenⅠ,CollagenⅢ,α-SMA and core proteinβ-catenin and p-GSK-3β/GSK-3βin Wnt/β-catenin signaling pathway in rat lung tissues decreased after H
作者
郑鑫
纵加强
王佳新
陈旭昕
韩志海
ZHENG Xin;ZONG Jiaqiang;WANG Jiaxin;CHEN Xuxin;HAN Zhihai(Navy Clinical College,the Fifth School of Clinical Medicine,Anhui Medical University,Hefei 230032,Anhui Province,China;Department of Pulmonary and Critical Care Medicine,the Sixth Medical Center,Chinese PLA General Hospital,Beijing 100048,China)
出处
《解放军医学院学报》
CAS
北大核心
2023年第12期1391-1398,1409,共9页
Academic Journal of Chinese PLA Medical School
基金
军队课题(BHJ16J011)。
