丙泊酚通过Fox O1/TXNIP信号通路介导自噬对高糖诱导心肌细胞损伤的作用  

Effects of propofol on high glucose induced myocardial cell injury through autophagy mediated by Fox O1/TXNIP signaling pathway

作　　者：杨育明 彭粤 韩红 任从才 赵丽霞 YANG Yu-ming;PENG Yue;HAN Hong;REN Cong-cai;ZHAO Li-xia(Department of Anesthesiology,The Eighth Affiliated Hospital of Sun Yat-sen University(Futian,Shenzhen),Shenzhen 518000,Guangdong Province,China)

机构地区：[1]中山大学附属第八医院(深圳福田)麻醉科,广东深圳518000

出　　处：《中国临床药理学杂志》2024年第16期2344-2348,共5页The Chinese Journal of Clinical Pharmacology

基　　金：广东省基础与应用基础研究基金资助项目(21201910240001116)。

摘　　要：目的探讨丙泊酚(Pro)通过叉头盒蛋白O1(Fox O1)/硫氧还蛋白相互作用蛋白(TXNIP)信号通路介导自噬对高糖(HG)诱导心肌细胞损伤的影响。方法将H9c2细胞分为空白组(5.5 mmol·L^(-1)葡萄糖)、高葡萄糖(HG)组(30 mmol·L^(-1)葡萄糖)、HG+Pro组(30 mmol·L^(-1)葡萄糖+25μmol·L^(-1)Pro)、HG+Pro+阴性对照(OE NC)组(先转染OE NC,再以30 mmol·L^(-1)葡萄糖+25μmol·L^(-1)Pro处理)、HG+Pro+Fox O1过表达质粒(Fox O1-OE)组(先转染Fox O1-OE,再以30 mmol·L^(-1)葡萄糖+25μmol·L^(-1)Pro处理)。用细胞计数试剂盒-8(CCK-8)法、原位末端转移酶标记技术(TUNEL)法、酶联免疫吸附试验(ELISA)、透射电镜、蛋白质印迹法(Western blot)分别检测细胞存活率、凋亡率、上清液中超氧化物歧化酶(SOD)、丙二醛(MDA)水平以及自噬体、Fox O1/TXNIP及自噬蛋白表达水平变化。结果空白组、HG组、HG+Pro组、HG+Pro+OE NC组、HG+Pro+Fox O1-OE组细胞活力分别为(100.00±0.00)%、(48.15±4.82)%、(79.66±7.98)%、(78.89±7.91)%和(49.22±4.93)%,凋亡率分别为(12.04±1.21)%、(42.34±4.25)%、(26.22±2.63)%、(27.02±2.71)%和(38.29±3.86)%,SOD水平分别为(62.24±6.25)、(28.21±2.85)、(55.37±5.58)、(55.09±5.53)和(30.66±3.08)U·mg^(-1)pro,MDA水平分别为(0.38±0.04)、(1.43±0.15)、(0.67±0.07)、(0.72±0.08)和(1.34±0.14)U·mg^(-1)pro,自噬体数量分别为(6.24±0.63)、(13.05±1.32)、(8.31±0.84)、(8.55±0.86)和(12.22±1.23)个,磷酸化Fox O1(p-Fox O1)/Fox O1比值分别为0.34±0.04、0.86±0.09、0.48±0.05、0.43±0.05和0.74±0.08,TXNIP表达分别为0.24±0.03、0.62±0.08、0.38±0.04、0.36±0.04和0.58±0.06,Bcelin-1表达分别为1.12±0.12、0.53±0.06、1.02±0.11、1.05±0.11和0.62±0.07,p62表达分别为0.56±0.06、1.56±0.16、0.82±0.09、0.86±0.09和1.44±0.15。HG组的上述指标与空白组比较,HG+Pro组的上述指标与HG组比较,HG+Pro+Fox O1-OE组的上述指标与HG+Pro+OE NC组比较,在统计学上差异均有统计学差异(均P<0.05)Objective To investigate the impact of propofol(Pro)on high glucose(HG)-induced myocardial cell injury through autophagy mediated by forkhead box O1(Fox O1)/thioredoxininteracting protein(TXNIP)signaling pathway.Methods H9c2 cells were divided into blank group(5.5 mmol·L^(-1)glucose),high glucose(HG)group(30 mmol·L^(-1)glucose),HG+Pro group(30 mmol·L^(-1)glucose+25μmol·L^(-1)Pro),HG+Pro+negative control(OE NC)group(first transfected with OE NC,then treated with 30 mmol·L^(-1)glucose+25μmol·L^(-1)Pro),HG+Pro+Fox O1 overexpression plasmid(Fox O1-OE)group(first transfected with Fox O1-OE,then treated with 30 mmol·L^(-1)glucose+25μmol·L^(-1)Pro).Cell counting kit-8(CCK-8)method,Td T-mediated d UTP nick end labeling(TUNEL)method,enzyme-linked immunosorbent assay(ELISA),transmission electron microscope and Western blot were applied to detect the cell survival rate,apoptosis rate,superoxide dismutase(SOD)and malondialdehyde(MDA)levels in the supernatant,and the changes in Autophagosome,Fox O1/TXNIP and autophagy protein expression levels.Results The cell viabilities of blank group,HG group,HG+Pro group,HG+Pro+OE NC group and HG+Pro+Fox O1-OE group were(100.00±0.00)%,(48.15±4.82)%,(79.66±7.98)%,(78.89±7.91)%and(49.22±4.93)%,respectively;the apoptosis rates were(12.04±1.21)%,(42.34±4.25)%,(26.22±2.63)%,(27.02±2.71)%,(38.29±3.86)%,respectively;SOD levels were(62.24±6.25),(28.21±2.85),(55.37±5.58),(55.09±5.53),(30.66±3.08)U·mg^(-1)pro,respectively;MDA levels were(0.38±0.04),(1.43±0.15),(0.67±0.07),(0.72±0.08)and(1.34±0.14)U·mg^(-1)pro,respectively;the number of autophagosomes was 6.24±0.63,13.05±1.32,8.31±0.84,8.55±0.86 and 12.22±1.23,respectively;phosphorylated Fox O1(p-Fox O1)/Fox O1 ratios were expressed as 0.34±0.04,0.86±0.09,0.48±0.05,0.43±0.05 and 0.74±0.08;TXNIP were expressed as0.24±0.03,0.62±0.08,0.38±0.04,0.36±0.04 and 0.58±0.06;Bcelin-1 were expressed as 1.12±0.12,0.53±0.06,1.02±0.11,1.05±0.11 and 0.62±0.07;p62 were expressed as 0.56±0.06,1.56±0.16,0.82±0.09

关 键 词：丙泊酚 叉头盒蛋白O1/硫氧还蛋白相互作用蛋白信号通路 自噬 高糖 心肌细胞损伤 

分 类 号：R972[医药卫生—药品]