摘要
目的观察氯沙坦通过高迁移率族蛋白B1(HMGB1)对热打击大鼠肝细胞中炎症反应的影响。方法体内实验中,将大鼠随机分为对照组(不接受热打击)、中暑组(中暑模型建立后腹腔注射生理盐水)和中暑氯沙坦组(中暑模型建立后腹腔注射氯沙坦50 mg/kg)。于模型建立9 h后取大鼠血清及肝组织,检测血清谷丙转氨酶(ALT)、肝组织髓过氧化物酶(MPO)、HMGB1水平及肝组织白介素(IL)-1β、IL-18水平,并观察肝组织病理形态学的变化。体外实验中,将大鼠HBL3A肝细胞系分为对照组(不接受热打击)、热打击组及热打击氯沙坦组(热打击后在上清中加入10μmol/L氯沙坦)。于模型建立9 h后检测细胞活力,上清乳酸脱氢酶及HMGB1水平,肝细胞胞质及总HMGB1表达,活化的半胱天冬酶-1表达,上清IL-1β、IL-18及细胞内活性氧簇水平。并分别观察添加梯度浓度的重组HMGB1和0.2 mmol/L H2O2对氯沙坦干预的热打击肝细胞上清IL-1β、IL-18及HMGB1水平的影响。结果体内实验中,与中暑组比较,中暑氯沙坦组血清ALT、肝组织MPO、血清HMGB1、肝组织IL-1β和IL-18水平均明显下降,差异有统计学意义(P<0.05);体外实验中,与热打击组比较,热打击氯沙坦组胞质HMGB1,上清HMGB1、IL-1β和IL-18水平均明显下降,细胞存活率上升,差异有统计学意义(P<0.05)。加入HMGB1抑制剂后,热打击肝细胞的IL-1β及IL-18水平均下降;加入HMGB1后,氯沙坦干预的热打击肝细胞的IL-1β与IL-18水平均上升(P<0.05)。氯沙坦干预后热打击肝细胞中的活性氧簇水平降低,加入H2O2后,氯沙坦干预的热打击肝细胞上清HMGB1水平增高(P<0.05)。结论氯沙坦可通过HMGB1抑制热打击大鼠肝细胞中的炎症损伤。
Objective To explore the function of losartan on heat-stress induced high-mobility group protein B1(HMGB1)mediated inflammatory damage in the hepatocytes.Methods Rats were randomized into three groups:sham group(without heat stress),heatstroke group(heatstroke induction followed by i.p.injection of normal saline)and heatstroke+losartan group(heatstroke induction followed by i.p.injection of 50 mg/kg losartan).The serum and liver tissue were harvested nine hours after heatstroke to invest the serum alanine aminotransferase(ALT)levels,liver myeloperoxidase(MPO)levels,liver pathological morphology,serum HMGB1 levels as well as the expression of interleukin(IL)-1βand IL-18 in the liver.In vitro,the HBL3 A hepatocyte cell lines were divided into the sham group(without heat stress),heat stress group and heat stress+losartan group(10μmol/L losartan added into the supernatant after heat stress).Nine hours after heat stress,the cell viability and the levels of supernatant lactate dehydrogenase,supernatant HMGB,cytoplasm and total HMGB1 were all examined.Besides,the level of activated caspase-1 in hepatocytes,supernatant IL-1βand IL-18,as well as the cellular level of reactive oxygen species(ROS),were detected.The effects of recombinant HMGB1 with concentration gradients and 0.2 mmol/L hydrogen peroxide on the levels of IL-1β,IL-18 and HMGB1 in the heat stress hepatocytes treated by losartan were observed.Results In vivo,liver damage occurred in the rats of the heatstroke group.Compared with the heatstroke group,the levels of serum ALT,liver MPO,serum HMGB1,liver tissue IL-1βand IL-18 decreased in the heatstroke+losartan group(P<0.05).In vitro,hepatocytes in the heat stress group were apparently damaged.Compared with the heat stress,the levels of cytoplasmic HMGB1,supernatant HMGB1,IL-1βand IL-18 decreased in the heat stress+losartan group,and the cell survival rate increased(P<0.05).In addition,the HMGB1 inhibitor also reduced the levels of IL-1βand IL-18 in heat stress group hepatocytes.And the supplementation with HMGB1 increased the levels of IL-1βand IL-18 in heat stress hepatocytes treated by losartan(P<0.05).Losartan reduced the level of reactive oxygen species in heat stress hepatocytes,and the supplementation with hydrogen peroxide increased the level of HMGB1 in heat stress hepatocytes treated by losartan(P<0.05).Conclusion Losartan decreased the heat-stress induced HMGB1 mediated inflammatory damage in the hepatocytes.
作者
张明
杨经文
陈怀生
李悦
施学智
梁泳欣
童华生
Zhang Ming;Yang Jing-Wen;Chen Huai-Sheng;Li Yue;Shi Xue-Zhi;Liang Yong-Xin;Tong Hua-Sheng(Department of Critical Care Medicine,the Sixth Affiliated Hospital of Guangzhou Medical University,Qingyuan People's Hospital,Qingyuan 511500,Guangdong,China;Department of Intensive Care Medicine,Shenzhen People’s Hospital,Shenzhen 518020,Guangdong,China;Department of Intensive Care Unit,General Hospital of Southern Theatre Command of PLA,Key Laboratory of Trauma and Tissue Repair of Tropical Area of PLA,Guangzhou 510010,China)
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2020年第3期285-291,共7页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金(81671896)
军事医学创新工程专项(18CXZ032)
广东省科技计划项目(2017A020215055)
清远市科技计划项目(2018B033)。
关键词
中暑
肝损伤
氯沙坦
高迁移率族蛋白B1
heat stroke
liver injury
losartan
high-mobility group protein B1
