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血必净减轻脓毒症肝细胞损伤的作用及机制研究

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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(40 μg·mL-1LPS)、血必净组(稀释25倍的血必净)、抑制药组(40 µg·mL-1 LPS+5 µmol·L-1 Nrf2 抑制药 ML385)、激活药组[40 μg·mL-1 LPS+15μg·mL-1萝卜硫素(Nrf2激活药)]、血必净+激活药组(40 μg·mL-1 LPS+稀释25倍的血必净+15 μg·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.
Effects of Xuebijing on reducing hepatocyte injury in sepsis
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(40 µg·mL-1 LPS),Xuebijing group(25 times diluted Xuebijing),inhibitor group(40 μg·mL-1 LPS+5 µg·mL-1 Nrf2 inhibitor ML385),activator group(40 μg·mL-1 LPS+15 µg·mL-1 Nrf2 activator sulforaphane),Xuebijing+activator group(40 μg·mL-1 LPS+25 times diluted Xuebijing+15 pg·mL-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 enhancing antioxidant system,and the combined treatment effect is better than Xuebijing alone.

Xuebijing injectionsepsiskidney injurynuclear factor E2 related factor 2activatoroxidative stress

周拓、丁锋、赵登峰

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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

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

2024

中国临床药理学杂志
中国药学会

中国临床药理学杂志

CSTPCD北大核心
影响因子:1.91
ISSN:1001-6821
年,卷(期):2024.40(16)
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