紫丁香苷对LPS/D-GalN诱导的急性肝损伤的保护作用研究

Protective effect of syringin on LPS/D-GalN induced acute liver injury

旨在探究紫丁香苷(syringin,SY)对脂多糖(LPS)联合D-半乳糖胺盐酸盐(D-GalN)、(LPS/D-GalN)诱导的小鼠急性肝损伤的保护作用及其潜在机制。将40只昆明鼠随机分为4组,分别为正常对照组(NC)、LPS/D-GalN模型(LD)组、SY低剂量(LSY+LD)组、SY高剂量(HSY+LD)组,每组10只。LSY+LD组、HSY+LD组采用腹腔注射方式给予SY(25、50 mg/kg),NC组和LD组分别腹腔注射等量的生理盐水,连续3 d,在第3天预防性给药1 h后,腹腔注射LPS/D-GalN建立急性药物肝损伤模型。造模6 h后,试验小鼠眼静脉取血,处死小鼠;检测小鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)的活性;检测肝匀浆中超氧化物歧化酶(SOD)、谷胱甘肽还原酶(GSH)、过氧化氢酶(CAT)的水平,观察肝脏氧化应激的水平;制作肝组织切片,采用苏木精伊红(HE)观察肝脏的组织变化;利用免疫组化观察激活的核转录因子p65(NF-κBp65)入核状态;ELISA法检肝匀浆IL-6、IL-1β、TNF-α炎性因子的水平。结果表明:与NC组相比,LD组AST、ALT的水平显著增高(P<0.01),肝血管严重出血,肝脏大面积实质性坏死,胞核碎裂、崩解,肝脏中SOD、GSH、CAT水平显著降低(P<0.05),NF-κBp65核转移率提高,肝脏中IL-6,IL-1β、TNF-α炎性因子水平显著升高(P<0.01);SY组可显著降低小鼠血清中AST、ALT的活性(P<0.05),肝组织病理变化明显减轻,肝脏SOD、CAT的活性与GSH的水平提高(P<0.05);NF-κBp65核转移发生抑制;IL-6,IL-1β、TNF-α水平显著降低(P<0.05)。结果表明SY(25、50 mg/kg)对LPS/D-GalN诱导的小鼠肝脏损伤具有保护作用且具有剂量依赖性,其作用机制与SY可提高抗氧化酶活性,抑制炎性因子的产生有关。

This experiment was to explore the protective effect and potential mechanism of syringin(SY)on LPS/D-GalN induced acute liver injury in mice.40 Kunming mice were used and randomly divided into four groups:the normal control group(NC),the LPS/D-GalN model(LD)group,the SY low dose(LSY+LD)group and the SY high dose(HSY+LD)group,with 10 mice in each group.The LSY+LD group and the HSY+LD group were given SY(25 and 50 mg/kg,respectively)by intraperitoneal injection.The NC group and the LD group were intraperitoneally injected with the same amount of normal saline for 3 days.One hour after the preventive administration on the third day,LPS/D-GalN was intraperitoneally injected to establish a model of acute drug-induced liver injury.6 hours after modeling,blood samples were collected from the ocular vein of the experimental mice which were then killed.Then,the activities of serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were detected,and the levels of superoxide dismutase(SOD),glutathione reductase(GSH)and catalase(CAT)in liver homogenate were detected to observe the level of oxidative stress in the liver.Next,liver tissue sections were made and stained with hematoxylin and eosin(HE).The immunohistochemical sections of NF-κBp65 were made to observe activated NF-κBp65 in the nucleus state.Finally,the IL-6 and IL-1βand TNF-α Levels of inflammatory factors in liver homogenate were detected by ELISA.The results showed that the levels of AST and ALT in the LD group were significantly higher than those in the NC group(P<0.01).Severe hemorrhage of the hepaticducts and blood vessels,the massive substantive necrosis of the liver,and the fragmentation and disintegration of nucleus were observed.The levels of SOD,GSH and CAT in the liver decreased significantly(P<0.05).The amount of NF-κBp65 nucleus increased.The inflammatory factors IL-6 IL-1β,and TNF-α in the liver increased significantly(P<0.01).The SY group had a significant reduction in the activities of AST and ALT in the serum(P<0.05).The pathological changes in the liver tissue were significantly reduced and the activities of SOD and CAT in the liver and the level of GSH were significantly increased(P<0.05).NF-κBp65 nuclear transfer was inhibited,and the IL-6 and IL-1β,TNF-α Levels were significantly reduced(P<0.05).The above results indicated that SY(25 and 50 mg/kg)had a protective effect on LPS/D-GalN induced liver injury in mice and it was dose-dependent.Its mechanism was similar to that of SY increasing the activity of antioxidant enzymes,and was related to the inhibition of production of inflammatory factors.