«上一篇/Previous Article|本期目录/Table of Contents|下一篇/Next Article»

《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:

37

期数:

2020年4

页码:

327-331

栏目:

基础研究

出版日期:

2020-04-05

文章信息/Info

Title:

Protective effect of mangiferin on acute liver failure in mice

作者:

徐　建¹; 柳晓娟¹; 吕庆芝¹; 余　红¹; 程卫光¹; 姬明丽²

（1.光山县人民医院儿科，河南　光山　465450；2.新乡医学院基础医学院，河南　新乡　453003）

Author(s):

XU Jian¹; LIU Xiaojuan¹; LYU Qingzhi¹; YU Hong¹; CHENG Weiguang¹; JI Mingli²

(1.Department of Pediatrics,Guangshan People′s Hospital,Guangshan 465450,Henan Province,China;2.School of Basic Medicine,Xinxiang Medical University,Xinxiang 453003,Henan Province,China)

关键词:

急性肝衰竭; 芒果苷; 肝功能; 炎症反应; 细胞凋亡

Keywords:

acute liver failure; mangiferin; liver function; inflammatory reaction; cell apoptosis

分类号:

R575.3

DOI:

10.7683/xxyxyxb.2020.04.006

文献标志码:

A

摘要:

目的　探讨芒果苷预处理对D-氨基半乳糖（D-GalN）/脂多糖（LPS）诱导的急性肝衰竭（ALF）小鼠的保护作用及其可能的机制。方法　75只C57BL6小鼠适应性喂养3 d后按随机数字表法分为对照组、模型组、芒果苷组，每组25只。芒果苷组小鼠给予芒果苷50 mg·kg^-1（溶解于0.1 mL磷酸盐缓冲溶液）灌胃，对照组、模型组小鼠灌胃同体积的磷酸盐缓冲溶液，每日1次，连续3 d。最后1次给药后2 h，模型组、芒果苷组小鼠腹腔注射D-GalN（700 mg·kg^-1）和 LPS（10 μg·kg^-1）溶液（溶解于0.1 mL磷酸盐缓冲溶液），建立ALF模型，对照组小鼠腹腔注射0.1 mL磷酸盐缓冲溶液。观察3组小鼠48 h生存率；造模后48 h，存活小鼠给予30 g·L^-1戊巴比妥钠2 mL·kg^-1腹腔注射麻醉，摘眼球取血，使用全自动生物化学分析仪测定血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、总胆汁酸（TBA）、总胆红素（TBIL）、直接胆红素（DBIL）水平，采用酶联免疫吸附法检测血清白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）水平；取血后处死小鼠，采集肝组织。苏木精-伊红染色观察小鼠肝组织病理学改变，末端脱氧核苷酰基转移酶介导性dUTP切口末端标记法测定肝细胞凋亡指数（AI），Western blot法检测小鼠肝细胞中核因子-κB（NF-κB） p65蛋白及肝组织中Bcl-2和Bax蛋白表达。结果　小鼠腹腔注射D-GalN和 LPS后48 h 内，对照组、模型组和芒果苷组小鼠生存率分别为100.0%（25/25只）、32.0%（8/25只）、76.0%（19/25只）；模型组小鼠生存率显著低于对照组，芒果苷组小鼠生存率显著高于模型组（P<0.01）。模型组小鼠血清ALT、AST、TBA、TBIL、DBIL水平显著高于对照组（P<0.01），芒果苷组小鼠血清ALT、AST、TBA、TBIL、DBIL水平显著低于模型组（P<0.01）。对照组小鼠肝小叶形态结构正常；模型组小鼠肝小叶结构严重破坏，肝细胞大片状坏死或呈散在性分布，细胞结构模糊不清，并伴炎性细胞浸润；与模型组相比，芒果苷组小鼠肝细胞坏死与炎性细胞浸润明显减轻。模型组小鼠血清IL-1β、TNF-α、IL-6水平显著高于对照组（P<0.01），芒果苷组小鼠血清IL-1β、TNF-α、IL-6水平显著低于模型组（P<0.01）。对照组、模型组和芒果苷组小鼠肝细胞AI分别为（0.08±0.02）%、（0.26±0.07）%和（0.15±0.04）%；模型组小鼠肝细胞AI高于对照组（P<0.01），芒果苷组小鼠肝细胞AI低于模型组（P<0.01）。模型组小鼠肝细胞内NF-κB p65蛋白相对表达量显著高于对照组（P<0.01），芒果苷组小鼠肝细胞内NF-κB p65蛋白相对表达量显著低于模型组（P<0.01）。模型组小鼠肝组织中Bcl-2蛋白相对表达量显著低于对照组（P<0.01），芒果苷组小鼠肝组织中Bcl-2蛋白相对表达量显著高于模型组（P<0.01）。模型组小鼠肝组织中Bax蛋白相对表达量显著高于对照组（P<0.01），芒果苷组小鼠肝组织中Bax蛋白相对表达量显著低于模型组（P<0.01）。结论　芒果苷对D-GalN/LPS诱导的ALF小鼠有保护作用，其机制可能与抑制炎症反应及肝细胞凋亡有关。

Abstract:

Objective　To investigate the protective effect and its possible mechanism of mangiferin pretreatment on acute liver failure (ALF) induced by D-galactosamine(D-GalN)/lipopolysaccharide (LPS) in mice.Methods　A total of 75 C57BL6 mice were randomly divided into control group,model group and mangiferin group according to the random number table method after adaptive feeding for 3 days,with 25 mice in each group.The mice in the mangiferin group were given mangiferin 50 mg·kg^-1 (dissolved in 0.1 mL phosphate buffered solution) by gavage,while the mice in the control group and model group were given the same volume of phosphate buffer solution once a day for three days.Two hours after the last administration,the mice in the model group and mangiferin group were treated with D-GalN (700 mg·kg^-1) and LPS (10 μg·kg^-1) solution (dissolved in 0.1 mL phosphate buffer solution) by peritoneal injection to establish ALF model,while the mice in the control group were treated with 0.1 mL phosphate buffer solution by peritoneal injection.The survival rate of mice in the three groups within 48 hours was observed.Forty-eight hours after the establishment of the models,the surviving mice were given 30 g·L^-1 pentobarbital sodium 2 mL·kg^-1 by intraperitoneal injection anesthesia,and the blood was taken from eyeballs.The levels of serum alanine aminotransferase (ALT),aspartate aminotransferase (AST),total bile acid (TBA),total bilirubin (TBIL) and direct bilirubin (DBIL) were measured with automatic biochemistry analyzer.The levels of serum interleukin-1β (IL-1β),tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were measured by enzyme linked immunosorbent assay.The mice were killed and the liver tissues were collected after blood collection.The histopathological changes of liver tissues of mice were observed by hematoxylin-eosin staining.The hepatocyte apoptosis index (AI) was measured by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay.The expression of nuclear factor-κB (NF-κB) p65 protein in hepatocytes and Bcl-2 and Bax protein in liver tissues were detected by western blot.Results　Within 48 hours after intraperitoneal injection of D-GalN and LPS,the survival rate of mice in the control group,model group and mangiferin group was 100.0% (25/25),32.0% (8/25) and 76.0% (19/25),respectively.The survival rate of mice in the model group was significantly lower than that in the control group,and the survival rate of mice in the mangiferin group was significantly higher than that in the model group (P<0.01).The levels of serum ALT,AST,TBA,TBIL and DBIL of mice in the model group were significantly higher than those in the control group (P<0.01),and the levels of serum ALT,AST,TBA,TBIL and DBIL of mice in the mangiferin group were significantly lower than those in the model group (P<0.01).The morphology and structure of hepatic lobule of mice in the control group were normal.In the model group,the structure of hepatic lobule was seriously damaged,the hepatocytes were necrotic or scattered,the cell structures were unclear and accompanied with inflammatory cell infiltration.Compared with the model group,the necrosis of hepatocytes and inflammatory cell infiltration were significantly reduced in the mangiferin group.The levels of serum IL-1β,TNF-α and IL-6 in the model group were significantly higher than those in the control group (P<0.01),while the levels of serum IL-1β,TNF-α and IL-6 in the mangiferin group were significantly lower than those in the model group (P<0.01).The AI of hepatocytes of mice in the control group,model group and mangiferin group was (0.08 ± 0.02)%,(0.26 ± 0.07)% and (0.15 ± 0.04)%,respectively.The AI of hepatocytes of mice in the model group was higher than that in the control group (P<0.01),while the AI of hepatocytes of mice in the mangiferin group was lower than that in the model group (P<0.01).The relative expression of NF-κB p65 protein in hepatocytes of mice in the model group was significantly higher than that in the control group (P<0.01),and the relative expression of NF-κB p65 protein in hepatocytes of mice in the mangiferin group was significantly lower than that in the model group (P<0.01).The relative expression of Bcl-2 protein in hepatic tissues of mice in the model group was significantly lower than that in the control group (P<0.01),and the relative expression of Bcl-2 protein in hepatic tissues of mice in the mangiferin group was significantly higher than that in the model group (P<0.01).The relative expression of Bax protein in hepatic tissues of mice in the model group was significantly higher than that in the control group (P<0.01),and the relative expression of Bax protein in hepatic tissues of mice in the mangiferin group was significantly lower than that in the model group (P<0.01).Conclusion　Mangiferin has protective effect on ALF induced by D-GalN/LPS in mice,and its mechanism may be related to inhibiting inflammatory response and hepatocyte apoptosis.

常用功能

更新日期/Last Update: 2020-04-05