丁酸钠对脂多糖联合D-氨基半乳糖诱导小鼠急性肝损伤的保护作用及其机制

doi:10.13481/j.1671-587X.20240615

摘要/Abstract

摘要：

目的探讨丁酸钠（NaB）脂多糖（LPS）联合D-氨基半乳糖（D-Gal）诱导小鼠急性肝损伤的保护作用，并阐明其作用机制。方法 30只雄性昆明小鼠随机分为对照组、模型组和NaB组，每组10只。NaB组小鼠给予200 mg·kg^-1·d^-1 NaB，对照组和模型组小鼠给予等体积无菌水。模型组和NaB组小鼠腹腔注射20 μg·kg^-1 LPS和600 mg·kg^-1 D-Gal诱导建立小鼠急性肝损伤模型。检测各组小鼠体质量和肝脏质量，计算肝脏指数。HE染色观察各组小鼠肝脏组织病理形态表现，试剂盒检测各组小鼠血清中丙氨酸氨基转移酶（ALT）和天门冬氨酸氨基转移酶（AST）活性及肝脏组织中总超氧化物歧化酶（T-SOD）和过氧化氢酶（CAT）活性及丙二醛（MDA）水平，Western blotting法检测各组小鼠肝脏组织中核因子E2相关因子2（Nrf2）和血红素加氧酶1（HO-1）蛋白表达水平。结果各组小鼠体质量比较差异均无统计学意义（P>0.05）；与对照组比较，模型组小鼠肝脏指数明显升高（P<0.01）；与模型组比较，NaB组小鼠肝脏指数明显降低（P<0.01）。HE染色观察，对照组小鼠肝脏组织结构正常，肝细胞边界清晰、大小一致，围绕中央静脉呈放射状均匀排列，且核位于细胞中央；模型组小鼠可见肝细胞排列紊乱，细胞肿胀，多发灶状肝细胞坏死，炎症细胞浸润及出血；与模型组比较，NaB组肝细胞形态结构得到改善，炎症浸润减少。与对照组比较，模型组小鼠血清中ALT和AST活性均明显升高（P<0.01）；与模型组比较，NaB组小鼠血清中ALT和AST活性均明显降低（P<0.05或P<0.01）。与对照组比较，模型组小鼠肝脏组织中T-SOD和CAT活性均明显降低（P<0.01），MDA水平明显升高（P<0.01）；与模型组比较，NaB组小鼠肝脏组织中T-SOD和CAT活性均明显升高（P<0.05或P<0.01），MDA水平明显降低（P<0.01）。Western blotting法检测，与对照组比较，模型组小鼠肝脏组织中Nrf2和HO-1蛋白表达水平均明显降低（P<0.05）；与模型组比较，NaB组小鼠肝脏组织中Nrf2和HO-1蛋白表达水平均明显升高（P<0.01）。结论 NaB对LPS/D-Gal诱导的小鼠急性肝损伤具有保护作用，其机制可能与NaB上调肝脏组织中Nrf2和HO-1蛋白表达和增加抗氧化酶活性，进而减轻肝脏氧化应激水平有关。

关键词: 丁酸钠, 急性肝损伤, 氧化应激, 核因子E2相关因子2, 血红素加氧酶1

Abstract:

Objective To discuss the protective effect of sodium butyrate（NaB） on acute liver injury in the mice induced by lipopolysaccharide（LPS） combined with D-galactosamine（D-Gal）， and to clarify its mechanism. Methods Thirty male Kunming mice were randomly divided into control group， model group， and NaB group， and there were 10 mice in each group. The mice in NaB group were given 200 mg·kg^-1·d^-1 NaB， while the mice in control group and model group were given an equal volume of sterile water. The mice in model group and NaB group were intraperitoneally injected with 20 μg·kg^-1 LPS and 600 mg·kg^-1 D-Gal to induce the acute liver injury models. The body weights and liver weights of the mice in various groups were detcted， and the liver index was calculated. HE staining was used to observe the pathomorphology of liver tissue of the mice in various groups； kits were used to detect the activities of alanine aminotransferase（ALT） and aspartate aminotransferase（AST） in serum， and the activities of total superoxide dismutase（T-SOD） and catalase（CAT）， and the levels of malondialdehyde（MDA） in liver tissue of the mice in various groups； Western blotting method was used to detect the expression levels of nuclear factor E2-related factor 2（Nrf2） and heme oxygenase-1（HO-1） proteins in liver tissue of the mice in various groups. Results There were no significant differences in body weights of the mice among various groups （P>0.05）. Compared with control group， the liver index of the mice in model group was significantly increased （P<0.01）. Compared with model group， the liver index of the mice in NaB group was significantly decreased （P<0.01）. The HE staining results showed that the liver tissue of the mice in control group exhibited normal structure， with clear boundaries of hepatocytes， consistent size， radially arranged around the central vein， and the nucleus located in the center of the cells； in model group， the arrangement of hepatocytes was disordered， the cells were swollen， there were multiple foci of hepatocellular necrosis， inflammatory cell infiltration， and hemorrhage； compared with model group， the cells in NaB group showed improved hepatocellular structure and reduced inflammatory infiltration. Compared with control group， the activities of ALT and AST in serum of the mice in model group were significantly increased （P<0.01）； compared with model group， the activities of ALT and AST in serum of the mice in NaB group were significantly decreased （P<0.05 or P<0.01）. Compared with control group， the activities of T-SOD and CAT in liver tissue of the mice in model group were significantly decreased （P<0.01）， and the level of MDA was significantly increased（P<0.01）； compared with model group， the activities of T-SOD and CAT in liver tissue of the mice in NaB group were significantly increased （P<0.05 or P<0.01）， and the level of MDA was significantly decreased（P<0.01）. The Western blotting results showed that compared with control group， the expression levels of Nrf2 and HO-1 proteins in liver tissue of the mice in model group were significantly decreased （P<0.05）； compared with model group， the expression levels of Nrf2 and HO-1 proteins in liver tissue of the mice in NaB group were significantly increased （P<0.01）. Conclusion NaB has a protective effect on LPS/D-Gal induced acute liver injury in the mice， and its mechanism may be related to the upregulation of the expressions of Nrf2 and HO-1 proteins and the increas of the activity of oxidant enzyme in liver tissue by NaB， thereby reduces the liver oxidative stress level of liver.

Key words: Sodium butyrate, Acute liver injury, Oxidative stress, Nuclear factor E2-related factor 2, Heme oxygenase-1

中图分类号:

R459.3

龙毅,游子怡,谭秀英,张柔,张钰浛,杨丽娜. 丁酸钠对脂多糖联合D-氨基半乳糖诱导小鼠急性肝损伤的保护作用及其机制[J]. 吉林大学学报(医学版), 2024, 50(6): 1614-1620.

Yi LONG,Ziyi YOU,Xiuying TAN,Rou ZHANG,Yuhan ZHANG,Lina YANG. Protective effect of sodium butyrate on acute liver injury in mice induced by lipopolysaccharide combined with D-galactosamine and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2024, 50(6): 1614-1620.

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Group	Body weight（m/g）	Liver index
Control	52.02±3.02	0.038±0.003
Model	49.88±2.89	0.074±0.010^*
NaB	47.09±2.36	0.056±0.009^△

Group	AST	ALT
Control	7.54±1.77	9.04±1.79
Model	38.51±6.56^*	106.66±31.98^*
NaB	20.37±3.00^△△	59.23±20.19^△

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