血必净减轻脓毒症肝细胞损伤的作用及机制研究  

Effects of Xuebijing on reducing hepatocyte injury in sepsis

作　　者：周拓 丁锋[1] 赵登峰[1] ZHOU Tuo;DING Feng;ZHAO Deng-feng(Department of Respiratory and Critical Care Medicine,First Affiliated Hospital of Nanyang Medical College,Nanyang 473000,Henan Province,China)

机构地区：[1]南阳医学高等专科学校第一附属医院呼吸与危重症医学科,河南南阳473000

出　　处：《中国临床药理学杂志》2024年第16期2349-2353,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的探讨血必净联合核因子E2相关因子2(Nrf2)激活药对脂多糖(LPS)诱导的肝细胞损伤的保护作用。方法将人正常肝细胞L-02随机分为对照组(正常培养)、LPS组(40μg·mL^(-1)LPS)、血必净组(稀释25倍的血必净)、抑制药组(40μg·mL^(-1)LPS+5μmol·L^(-1)Nrf2抑制药ML385)、激活药组[40μg·mL^(-1)LPS+15μg·mL^(-1)萝卜硫素(Nrf2激活药)]、血必净+激活药组(40μg·mL^(-1)LPS+稀释25倍的血必净+15μg·mL^(-1)萝卜硫素);处理24 h后,用蛋白质印迹法检测细胞核Nrf2,用5-乙炔基-2’-脱氧尿苷(Edu)实验检测细胞增殖情况,用流式细胞术检测细胞凋亡情况,用荧光探针法检测各组细胞活性氧(ROS)水平。结果对照组、LPS组、血必净组、抑制药组、激活药组、血必净+激活药组细胞核Nrf2蛋白表达水平分别为0.83±0.08、0.40±0.04、0.59±0.07、0.26±0.04、0.90±0.10和1.24±0.13,Edu阳性细胞率分别为(42.99±4.37)%、(16.30±1.42)%、(30.08±2.10)%、(14.67±1.27)%、(28.66±2.54)%和(38.31±2.54)%,细胞凋亡率分别为(4.39±0.42)%、(25.17±3.70)%、(14.44±0.99)%、(28.14±1.26)%、(17.99±1.30)%和(10.50±0.86)%,ROS平均光密度值分别为1.00±0.04、3.48±0.34、1.88±0.15、4.25±0.37、1.87±0.19和1.26±0.12;LPS组上述指标与对照组比较,在统计学上差异均有统计学意义(均P<0.05);血必净组、抑制药组、激活药组上述指标与LPS组比较,在统计学上差异均有统计学意义(均P<0.05);血必净+激活药组上述指标分别与血必净组、激动剂组比较,在统计学上差异均有统计学意义(均P<0.05)。结论血必净联合Nrf2激活药可通过增强抗氧化系统减轻LPS诱导的肝细胞损伤,其联合治疗效果优于血必净单独用。Objective To investigate the protective effect of Xuebijing combined with nuclear factor E2-related factor 2(Nrf2)activator on lipopolysaccharide(LPS)-induced hepatocyte injury.Methods Human normal liver cells L-02 were randomly divided into control group(normal culture),LPS group(40μg·m L^(-1)LPS),Xuebijing group(25 times diluted Xuebijing),inhibitor group(40μg·m L^(-1)LPS+5μg·m L^(-1)Nrf2 inhibitor ML385),activator group(40μg·m L^(-1)LPS+15μg·m L^(-1)Nrf2 activator sulforaphane),Xuebijing+activator group(40μg·m L^(-1)LPS+25 times diluted Xuebijing+15μg·m L^(-1)Nrf2 activator sulforaphane).After treatment for 24 h,the expression of nuclear Nrf2 was detected by Western blot assay;cell proliferation was detected by 5-ethynyl-2'-deoxyuridine(Edu)assay;cell apoptosis was detected by flow cytometry;reactive oxygen species(ROS)levels were detected by fluorescent probe.Results The nuclear Nrf2 protein levels of control group,LPS group,Xuebijing group,inhibitor group,activator group and Xuebijing+activator group were 0.83±0.08,0.40±0.04,0.59±0.07,0.26±0.04,0.90±0.10,1.24±0.13,respectively;the Edu positive cell rates were(42.99±4.37)%,(16.30±1.42)%,(30.08±2.10)%,(14.67±1.27)%,(28.66±2.54)%,(38.31±2.54)%,respectively;the apoptosis rates were(4.39±0.42)%,(25.17±3.70)%,(14.44±0.99)%,(28.14±1.26)%,(17.99±1.30)%,(10.50±0.86)%,respectively;the mean density values of ROS were 1.00±0.04,3.48±0.34,1.88±0.15,4.25±0.37,1.87±0.19,1.26±0.12,respectively.The above indexes of LPS group were significantly different from those of control group(all P<0.05);the above indexes of Xuebijing group,inhibitor group and activator group were compared with LPS group,respectively,and the differences were statistically significant(all P<0.05);the above indexes of Xuebijing+agonist group were compared with Xuebijing group or agonist group,respectively,and the differences were statistically significant(all P<0.05).Conclusion Xuebijing combined with Nrf2 activator can reduce LPS-induced hepatocyte injury by enhancin

关 键 词：血必净注射液 脓毒症 肾损伤 核因子E2相关因子2 激活药 氧化应激 

分 类 号：R28[医药卫生—中药学]