加味桃仁承气汤对机械通气相关性肺损伤大鼠细胞自噬的影响  

Effect of Jiawei Taoren Chengqi Decoction on autophagy in rats with mechanical ventilation-induced lung injury

作　　者：孙治霞[1] 王丽辉[1] 索红亮[2] 陈乾 SUN Zhixia;WANG Lihui;SUO Hongliang;CHEN Qian(Department of Critical Care Medicine,Henan Provincial Hospital of Traditional Chinese Medicine,Zhengzhou 450002,China;Department of Cardiology,Henan Provincial Hospital of Traditional Chinese Medicine,Zhengzhou 450002,China)

机构地区：[1]河南省中医院重症医学科,450002 [2]河南省中医院心病科,450002

出　　处：《天津医药》2023年第12期1349-1355,共7页Tianjin Medical Journal

基　　金：河南省卫生健康委国家中医临床研究基地科研专项(2021JDZX2033)。

摘　　要：目的 探究加味桃仁承气汤通过腺苷酸活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路对机械通气相关性肺损伤(VILI)大鼠细胞自噬的影响。方法 将60只SPF级雄性SD大鼠随机分为假手术组(Sham组)、模型组(Model组)、加味桃仁承气汤低剂量组(中药-L组,2.85 g/kg)、加味桃仁承气汤高剂量组(中药-H组,8.55 g/kg)和加味桃仁承气汤高剂量+AMPK抑制剂Compound C组(中药-H+CC组,加味桃仁承气汤8.55 g/kg+Compound C 250μg/kg),每组12只。于插管即刻和机械通气1 h、2 h与4 h时测定氧合指数(OI)。机械通气结束后收集肺泡灌洗液(BALF),酶联免疫吸附试验(ELISA)检测BALF中肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β、IL-18水平;处死大鼠,取肺组织测定湿/干重(W/D)比值;HE染色观察各组大鼠肺组织病理学改变并进行肺损伤评分;透射电镜观察肺泡上皮细胞形态。RT-qPCR检测大鼠肺组织中的AMPK、mTORC1 mRNA表达水平;Western blot检测大鼠肺组织中AMPK、p-AMPK、mTORC1、p-mTORC1和自噬相关蛋白的表达情况。结果 与Sham组相比,Model组大鼠肺组织病理损伤严重,肺W/D、BALF中TNF-α、IL-1β和IL-18水平、肺损伤评分、mTORC1 mRNA表达水平、pmTORC1蛋白表达升高(P<0.05);机械通气2 h、4 h时OI,肺组织AMPK mRNA表达水平,p-AMPK、LC3-Ⅱ/LC3-Ⅰ、Beclin-1蛋白表达降低(P<0.05)。与Model组相比,中药-H组大鼠肺组织病理损伤减轻,相关指标的变化趋势与上述相反(P<0.05),肺泡上皮细胞自噬小体增多。Compound C减弱了加味桃仁承气汤对VILI大鼠的保护作用(P<0.05)。结论 加味桃仁承气汤可能通过激活AMPK/mTOR信号通路促进细胞自噬,减轻大鼠VILI。Objective To explore the impact of Jiawei Taoren Chengqi Decoction on autophagy in rats with mechanical ventilation-induced lung injury(VILI)through adenylate-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)signaling pathway.Methods Sixty SPF male rats were randomly grouped into the sham operation group,the model group,the low-dose Jiawei Taoren Chengqi Decoction group(TCM-L group,2.85 g/kg),the high-dose Jiawei Taoren Chengqi Decoction group(TCM-H group,8.55 g/kg)and the high-dose Jiawei Taoren Chengqi Decoction+AMPK inhibitor Compound C group(TCM-H+CC group,Jiawei Taoren Chengqi Decoction 8.55 g/kg+Compound C 250μg/kg),12 in each group.Oxygenation index(OI)was measured immediately after intubation and at 1 h,2 h and 4 h after mechanical ventilation.Bronchoalveolar lavage fluid(BALF)was collected after mechanical ventilation,levels of tumor necrosis factor(TNF)-α,interleukin(IL)-1βand IL-18 in BALF were detected by enzyme-linked immunosorbent assay(ELISA).Rats were sacrificed,and lung tissue was taken to measure the wet/dry weight(W/D)ratio.HE staining was used to observe pathological changes of lung tissue in each group of rats.Lung injury scores were carried out.Morphology of alveolar epithelial cells was observed by transmission electron microscopy.RT-qPCR was applied to detect mRNA expression levels of AMPK and mTORC1 in rat lung tissue.Western blot assay was applied to detect expression levels of AMPK,p-AMPK,mTORC1,p-mTORC1 and autophagy-related proteins in rat lung tissue.Results Compared with the sham group,the pathological damage of lung tissue was serious,lung W/D,levels of TNF-α,IL-1βand IL-18 in BALF,lung injury score,mTORC1 mRNA expression level,and p-mTORC1 protein expression were increased in the model group(P<0.05).OIvalues at 2 h and 4 h of mechanical ventilation,AMPK mRNA expression level,p-AMPK,LC3-II/LC3-I and Beclin-1 protein expression in lung tissue were decreased(P<0.05).Compared with the model group,the pathological damage of lung tissue was alleviated in the Chines

关 键 词：呼吸机相关性肺损伤 桃核承气汤 自噬 AMP活化蛋白激酶类 TOR丝氨酸-苏氨酸激酶 

分 类 号：R563[医药卫生—呼吸系统]