甘草酸通过激活核因子E2相关因子2/血红素加氧酶1通路减轻急性胰腺炎氧化应激

Liquiritigenin attenuates the oxidative stress in acute pancreatitis by activating Nrf2/HO-1 pathway

目的探讨甘草酸(liquiritigenin,Liq)对急性胰腺炎(acute pancreatitis,AP)小鼠模型的疗效及作用机制。方法24只C57BL/6小鼠随机分为对照组(CON组)、甘草酸对照组(Liq组)、AP组、AP+甘草酸治疗组(AP+Liq组),6只/组。腹腔注射两次L-精氨酸(4 g/kg)建立AP小鼠模型,两次注射间隔1 h;CON组注射等体积生理盐水;Liq组及AP+Liq组于造模前1 h通过腹腔注射甘草酸(30 mg/kg)。造模后72 h处死小鼠采集标本,光学显微镜下观察胰腺组织病理改变,ELISA方法检测血清淀粉酶、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量,检测胰腺组织中丙二醛(MDA)及超氧化物歧化酶(SOD)含量,免疫荧光检测胰腺组织活性氧(ROS)表达,Western blot检测胰腺组织中核因子E2相关因子2(Nrf2)、血红素加氧酶1(HO-1)的蛋白表达。结果与CON组比较,AP组小鼠胰腺明显坏死,血清淀粉酶(U/L:107.10±19.29 vs.639.4±59.10)、TNF-α(pg/mL:12.77±1.06 vs.106.20±16.13)、IL-6(pg/mL:11.74±1.75 vs.105.21±11.42)、MDA(U/mg:0.81±0.18 vs.3.73±0.87)明显升高(P<0.01),SOD(U/mg:6.76±1.10 vs.3.02±0.88)明显下降(P<0.01),同时Nrf2/HO-1通路被明显抑制;与AP组比较,AP+Liq组胰腺组织损伤明显减轻,淀粉酶(U/L:216.80±44.73)、TNF-α(pg/mL:36.48±7.06)、IL-6(pg/mL:38.66±11.29)及MDA(U/mg:2.09±0.57)明显降低(P<0.01和P<0.05),SOD(U/mg:5.70±1.01)表达上调,同时Nrf2/HO-1通路表达上调;Liq组与CON组各指标比较差异无统计学意义(P>0.05)。结论甘草酸能够明显减轻AP,其机制可能是通过激活Nrf2/HO-1通路减轻AP氧化应激。

Objective To study the efficacy and mechanism of liquiritigenin(Liq)on acute pancreatitis(AP)mouse models.Methods Twenty-four C57BL/6 mice were randomly divided into control group(CON group),liquiritigenin control group(Liq group),acute pancreatitis group(AP group),acute pancreatitis+liquiritigenin treatment group(AP+Liq group),6 mice/per group.L-arginine(4 g/kg)was intraperitoneal injected twice to establish AP mouse models,with an interval of 1 hour between the two injections;the CON group was injected with equal volume of saline;the liquiritigenin(30 mg/kg)was injected 1 h before modeling in Liq group and AP+Liq group.Seventy-two hours after modeling,the mice were sacrificed and specimens were collected.The pathological and morphological changes of pancreatic tissue were observed under light microscope,serum amylase levels,inflammatory factors interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)content,malondialdehyde(MDA)and superoxide dismutase(SOD)content were detected by enzyme-linked immunosorbent assay(ELISA),reactive oxygen species(ROS)expression in pancreatic tissue was detcted by immunofluorescence,protein expression of nuclear factor erythroid 2-related factor 2(Nrf2)and heme oxygenase-1(HO-1)in pancreatic tissue was detcted by Western blot test.Results Compared with the CON group,the AP group mice showed obvious pancreatic necrosis,serum amylase(U/L:107.10±19.29 vs.639.40±59.10),TNF-α(pg/mL:12.77±1.06 vs.106.20±16.13),IL-6(pg/mL:11.74±1.75 vs.105.21±11.42),MDA(U/mg:0.81±0.18 vs.3.73±0.87)were significantly increased(P<0.01),SOD(U/mg:6.76±1.10 vs.3.02±0.88)decreased(P<0.01),and Nrf2/HO-1 pathway was significantly inhibited;Compared with the AP group,the pancreatic tissue damage in the AP+Liq group was significantly reduced,the levels of amylase(U/L:216.80±44.73),TNF-α(pg/mL:36.48±7.06),IL-6(pg/mL:38.66±11.29)and MDA(U/mg:2.09±0.57)were significantly reduced(P<0.01 and P<0.05),the expression of SOD(U/mg:5.70±1.01)was upregulated,and the expression of Nrf2/HO-1 pathway was up-regulated;There were no significant differences in the indexes between Liq group and CON group(P>0.05).Conclusions Liquiritigenin can significantly alleviate AP by the activation of Nrf2/HO-1 signaling pathway.