摘要
目的 探讨D-氨基半乳糖(D-GalN)/脂多糖(LPS)诱导急性肝衰竭(acute liver failure,ALF)大鼠中髓过氧化物酶(myeloperoxidase,MPO)和核因子E2相关因子2(Nrf2)/血红素氧合酶-1(HO-1)信号通路的变化及ALF的发生机制。方法 SD大鼠随机分为对照组和ALF组。ALF组:将D-GalN 800 mg/kg和LPS 8μg/只同时腹腔注射,于注射后6、12和24 h检测血清总胆红素(TBiL)、丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)含量。ELISA法检测血清MPO、肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)水平,比色法测定MPO活性,RT-PCR检测肝组织Nrf2和HO-1的mRNA水平,Western blot法检测肝组织MPO、Nrf2和HO-1蛋白水平。结果 与对照组比较,ALF组血清MPO水平于造模后6 h升高,12 h为最高,24 h开始下降(23.33±2.06 vs.33.00±3.16,65.75±7.02,53.92±5.63,P<0.05);血清TNF-α(11.30±3.26 vs.102.17±14.80,83.33±11.22,64.25±9.29,P<0.01)和IL-6(10.83±2.92 vs.89.25±10.86,77.33±8.02,65.58±7.31,P<0.01)于造模后6 h升高最明显,12 h后逐渐下降,差异均有统计学意义。ALF组6、12、24 h肝组织MPO活性高于对照组,差异均有统计学意义(15.5±1.51,28.08±4.65,22.92±1.93 vs.12.17±1.27,P<0.05);ALF组肝组织6、12、24 h Nrf2 mRNA(1.59±0.13,3.65±0.11,2.35±0.11 vs.1.04±1.01,P<0.01)和HO-1 mRNA(2.44±0.19,4.77±0.18,3.82±0.17 vs.1.12±0.06,P<0.01)水平高于对照组,差异均有统计学意义。ALF组肝组织MPO(4.10±0.70,9.77±1.15,7.23±0.40 vs.2.07±0.42,P<0.01)、Nrf2(4.03±0.80,9.03±0.50,6.07±0.47 vs.2.63±0.38,P<0.01)和HO-1(1.73±0.21,5.17±0.51,3.03±0.32 vs.0.97±0.21,P<0.01)蛋白表达于12 h达最高值,ALF组各时间点与对照组比较均差异有统计学意义。结论 MPO可能通过氧化应激和炎症反应影响Nrf2/HO-1信号通路,在ALF中发挥重要作用。
Objective To investigate the changes of myeloperoxidase(MPO) and NF-E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1) signaling pathway and the pathogenesis in D-galactosamine(D-GalN)/lipopolysaccharide(LPS)-induced acute liver failure(ALF) rats.Methods SD rats were randomly divided into control group and ALF model group.ALF group:total bilirubin(TbiL),alanine aminotransferase(ALT) and aspartate aminotransferase(AST) levels were detected at 6,12 and 24 h after intraperitoneal injection of D-GalN 800 mg/kg and LPS 8 μg/rat.MPO,TNF-α and IL-6 levels were detected by ELISA.MPO activity was detected by colorimetry.Levels of Nrf2 and HO-1 mRNA were measured by RT-PCR.The expression of MPO,Nrf2 and HO-1 proteins in liver tissues were detected by Western blot.Results Compared with the control group,the level of MPO in ALF group increased at 6 h,reached the highest level at 12 h after modeling,and began to decrease at 24 h(23.33±2.06 vs.33.00±3.16,65.75±7.02,53.92±5.63,P<0.05);TNF-α(11.30±3.26 vs.102.17±14.80,83.33±11.22,64.25±9.29,P<0.01) and IL-6(10.83±2.92 vs.89.25±10.86,77.33±8.02,65.58±7.31,P<0.01) were peaked at 6 h after modeling,and gradually decreased at 12 h,and the differences were statistically significant.The activity of MPO in liver tissue of ALF group was higher than that of control group at 6,12,24 h(15.5±1.51,28.08±4.65,22.92±1.93 vs.12.17±1.27,P<0.05),the differences were statistically significant;The mRNA levels of Nrf2 and HO-1 in liver tissue of ALF group were higher than those of control group,Nrf2 mRNA(1.59±0.13,3.65±0.11,2.35±0.11 vs.1.04±1.01,P<0.01),HO-1 mRNA(2.44±0.19,4.77±0.18,3.82±0.17 vs.1.12±0.06,P<0.01),the differences were statistically significant.The protein expression of MPO(4.10±0.70,9.77±1.15,7.23±0.40 vs.2.07±0.42,P<0.01),Nrf2(4.03±0.80,9.03±0.50,6.07±0.47 vs.2.63±0.38,P<0.01)and HO-1(1.73±0.21,5.17±0.51,3.03±0.32 vs.0.97±0.21,P<0.01)in liver tissue of ALF group reached the highest value at 12 h,there were significant differences between ALF and
作者
王柯尹
魏大海
邹卓林
孙丹凤
徐艳丽
Wang Ke-yin;Wei Da-hai;Zou Zhuo-lin;Sun Dan-feng;Xu Yan-li(Department of Infectious Disease,the First Hospital of Jiaxing,Jiaxing 314000,China)
出处
《中国急救医学》
CAS
CSCD
2022年第8期717-722,共6页
Chinese Journal of Critical Care Medicine
基金
嘉兴市科技计划项目(2021AD30154)
浙江省自然科学基金/探索项目(LY20C010004)。
