摘要
目的研究四氢姜黄素(tetrahydrocurcumin,THC)对脓毒血症小鼠心肌损伤的保护作用及其分子机制。方法将40只C57BL/6J小鼠随机分为4组:对照组、LPS组、LPS+THC组和EX527+LPS+THC组,每组10只。采用腹腔注射脂多糖(LPS,10 mg/kg)的方法构建小鼠脓毒血症心肌损伤模型。对照组和LPS组小鼠每日给予等量生理盐水灌胃处理一次;在LPS腹腔注射前7 d,LPS+THC组和EX527+LPS+THC组小鼠每日给予THC[100 mg/(kg·d)]灌胃处理一次;且EX527+LPS+THC组小鼠在药物灌胃处理的同时给予5 mg/kg的EX527(SIRT1信号特异性阻断剂)腹腔注射处理一次。待LPS注射6 h后,用小动物超声仪检测各组小鼠左心室射血分数(LVEF)和左心室短轴缩短率(LVFS);用酶标仪测定血清中CK、CK-MB和TnI的活性;用ELISA法检测心肌组织IL-1β和TNF-α表达水平;用免疫组化染色法检测心肌组织中性粒细胞浸润程度;用试剂盒检测心肌组织MDA含量和SOD活性;用TUNEL试剂盒检测心肌细胞凋亡水平;用DHE荧光探针检测心肌组织内活性氧簇产量;用Western blot检测SIRT1、Ac-foxo1和gp91^(phox)蛋白表达情况。结果与对照组相比,LPS组小鼠血清CK、CK-MB和TnI活性均显著升高,LVEF和LVFS均显著降低,心肌组织氧化应激水平显著升高,炎症反应明显增强,心肌细胞凋亡率显著增加,SIRT1蛋白表达明显下调,foxo1的乙酰化水平明显上调(均P<0.05)。与LPS组相比,LPS+THC组小鼠经THC预处理后可明显改善LPS引起的上述改变(P<0.05)。然而,THC的保护效果均可被EX527显著逆转(P<0.05)。结论四氢姜黄素可通过激活SIRT1信号通路发挥抗氧化应激与抗炎作用,进而减轻了脓毒血症小鼠的心肌损伤。
Objective To explore the protective effects of tetrahydrocurcumin(THC)against lipopolysaccharide(LPS)-induced myocardial injury and its underlying molecular mechanism.Methods Forty C57BL/6J mice were randomly divided into 4 groups(n=10 in each group):control group,LPS group,LPS+THC group and EX527+LPS+THC group.The mouse model of sepsis-associated cardiomyopathy was established by intraperitoneal injection of LPS(10 mg/kg).Seven days before LPS injection,mice in LPS+THC group and EX527+LPS+THC group were administered intragastrically with THC[100 mg/(kg·d)],while the mice in control group and LPS group were treated with same volume of saline solution.EX527(SIRT1 specific inhibitor,5 mg/kg)was given intraperitoneally at the same time of THC administration in EX527+LPS+THC group.Six hours after LPS injection,Vevo 770 ultrasound instrument was used to detect left ventricular ejection fraction(LVEF)and left ventricular fractional shortening(LVFS),plate analyzer was used to determine the activities of serum CK,CK-MB and TnI,ELISA kits were used to measure the expression levels of IL-1βand TNF-αin myocardium,immunohistochemical staining was used to detect the degree of neutrophil infiltration,special kits were used to estimate the contents of superoxide dismutase(SOD)and malondialdehyde(MDA)in myocardium,TUNEL staining was used to determine the cardiomyocyte apoptosis,DHE fluorescence probe was used to examine the production of reactive oxygen species(ROS),and Western blot assay was used to assess the protein expression levels.Results Compared with control group,the activities of serum CK,CK-MB and TnI were significantly raised,while LVEF and LVFS were significantly decreased in LPS group.Additionally,the levels of oxidative stress,the inflammatory response and the rate of cardiomyocyte apoptosis in myocardium were significantly increased in LPS group,the expression of SIRT1 was reduced,and the acetylation level of foxo1 was increased(P<0.01).Compared with LPS group,the above-mentioned changes were all obviously ame
作者
王伟
曾广伟
廉诚
刘慧
张晓东
安慧仙
高钊
方东
张艳涛
樊贝贝
孙梦娜
李炜
WANG Wei;ZENG Guangwei;LIAN Cheng;LIU Hui;ZHANG Xiaodong;AN Huixian;GAO Zhao;FANG Dong;ZHANG Yantao;FAN Beibei;SUN Mengna;LI Wei(Department of Cardiology,Xi’an International Medical Center Hospital,Xi’an 710100,China;Department of Cardiovascular Diseases,Second Affiliated Hospital,Air Force Medical University)
出处
《山西医科大学学报》
CAS
2021年第4期443-450,共8页
Journal of Shanxi Medical University
基金
陕西省重点研发计划项目(2019SF-202)
唐都医院创新发展基金项目(2018QYTS011)。
