摘要
参附注射液(Shenfu Injection,SFI)在脓毒症休克治疗方面起着重要作用,对于脓毒症肺损伤的治疗先前已有研究,但其具体作用机制尚不明确,该研究从SFI调控低氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)从而介导线粒体自噬方面,分别从临床、动物实验等探讨SFI对于脓毒症肺损伤的保护作用,选取郑州大学第一附属医院急诊重症监护病房(emergency intensive care unit,EICU)收治的脓毒症肺损伤确诊患者70例,检测患者第1、5天的白细胞介素(interleukin,IL)-6和肿瘤坏死因子(tumor necrosis factor,TNF)-α,同时采用实时荧光定量聚合酶链反应(real-time quantitative polymerasechain reaction,RT-qPCR)检测患者第5天的外周血单核细胞(peripheral blood mononuclear cell,PBMC)中缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)的表达。动物实验方面采用随机数表法将32只SPF级5~6周雄性C57BL/6J小鼠随机分为4组:假手术(sham)组(n=6),SFI+sham组(n=10),SFI+盲肠结扎穿刺(CLP)组(n=10),CLP组(n=6)。记录4组小鼠体质量及体温的变化、肺组织湿/干重(W/D)、肺组织病理损伤评分;随后通过RT-qPCR及Western blot(WB)检测4组小鼠肺组织中HIF-1α、线粒体DNA(mitochondrial DNA,mt-DNA)及自噬相关蛋白的表达。结果表明,参附注射液组患者血液及肺泡灌洗液炎症指标表达下降,外周血PBMC中HIF-1α的表达明显升高,且具有一定相关性。在小鼠实验中,SFI组小鼠体温、体质量变化好转,血清TNF-α降低,肺组织W/D显著减小,在肺组织苏木素-伊红(HE)染色中肺组织炎性渗出及肺泡完整性明显好转,RT-qPCR定量脓毒症肺损伤小鼠mtDNA表达明显下降,肺组织蛋白WB半定量发现线粒体动力蛋白线粒体转录因子A(mt-TFA)及线粒体功能蛋白细胞色素c氧化酶Ⅳ(cytochrome c oxidaseⅣ,COXⅣ)表达增加,HIF-1α及自噬相关蛋白表达增加。该研究初步明确SFI通过增加线粒体自噬功能,改善线粒体功能,减轻
Shenfu Injection(SFI)is praised for the high efficacy in the treatment of septic shock.However,the precise role of SFI in the treatment of sepsis-associated lung injury is not fully understood.This study investigated the protective effect of SFI on sepsis-associated lung injury by a clinical trial and an animal experiment focusing on the hypoxia-inducing factor-1α(HIF-1α)-mediated mitochondrial autophagy.For the clinical trial,70 patients with sepsis-associated lung injury treated in the emergency intensive care unit of the First Affiliated Hospital of Zhengzhou University were included.The levels of interleukin(IL)-6 and tumor necrosis factor(TNF)-αwere measured on days 1 and 5 for every patient.Real-time quantitative polymerase chain reaction(RT-qPCR)was performed to determine the mRNA level of hypoxia inducible factor-1α(HIF-1α)in the peripheral blood mononuclear cells(PBMCs).For the animal experiment,32 SPF-grade male C57BL/6J mice(5-6 weeks old)were randomized into 4 groups:sham group(n=6),SFI+sham group(n=10),SFI+cecal ligation and puncture(CLP)group(n=10),and CLP group(n=6).The body weight,body temperature,wet/dry weight(W/D)ratio of the lung tissue,and the pathological injury score of the lung tissue were recorded for each mouse.RT-qPCR and Western blot were conducted to determine the expression of HIF-1α,mitochondrial DNA(mt-DNA),and autophagy-related proteins in the lung tissue.The results of the clinical trial revealed that the SFI group had lowered levels of inflammatory markers in the blood and alveolar lavage fluid and elevated level of HIF-1αin the PBMCs.The mice in the SFI group showed recovered body temperature and body weight.lowered TNF-αlevel in the serum,and decreased W/D ratio of the lung tissue.SFI reduced the inflammatory exudation and improved the alveolar integrity in the lung tissue.Moreover,SFI down-regulated the mtDNA expression and upregulated the protein levels of mitochondrial transcription factor A(mt-TFA),cytochrome c oxidaseⅣ(COXⅣ),HIF-1α,and autophagy-related prote
作者
张娈娈
訾亚楠
张业鹏
裴辉
郑湘予
谢佳丰
徐东
朱志强
ZHANG Luan-luan;ZI Ya-nan;ZHANG Ye-peng;PEI Hui;ZHENG Xiang-yu;XIE Jia-feng;XU Dong;ZHU Zhi-qiang(Emergency Department,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China;Department of Pathology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)
出处
《中国中药杂志》
CSCD
北大核心
2023年第23期6492-6499,共8页
China Journal of Chinese Materia Medica
基金
河南省中医药科学研究专项(2022ZY1210)
河南省高等学校重点科研项目(22A320056)
2022睿E(睿意)急诊医学研究专项(22222012017)。
