消退素D1预处理对肝缺血再灌注损伤大鼠模型的保护作用及机制

王阳阳; 彭雪莹; 孟杰; 安博然; 贺文娟; 鲁素彩; 陈岩; 孔立文; 牛川; 刘丛; 黄薇; 侯英键

doi:10.3969/j.issn.1001-5256.2021.06.030

消退素D1预处理对肝缺血再灌注损伤大鼠模型的保护作用及机制

DOI: 10.3969/j.issn.1001-5256.2021.06.030

王阳阳¹,
彭雪莹^{1, 2},
孟杰^1, ,,
安博然¹,
贺文娟³,
鲁素彩¹,
陈岩¹,
孔立文⁴,
牛川³,
刘丛⁵,
黄薇¹,
侯英键¹

1.
河北大学附属医院消化科，河北保定 071000
2.
河北大学临床医学院，河北保定 071000
3.
河北省易县人民医院内科，河北保定 074200
4.
河北容城县中医医院内科，河北保定 071700
5.
保定市第一中医院内科，河北保定 071000

基金项目:

国家自然科学基金 (812000078);

保定市2019年度科技计划项目 (1951ZF092);

河北省卫生和计划生育委员会2018年医学科学研究重点课题计划项目 (20180704);

河北大学医学学科培育项目 (2020A12);

河北大学附属医院青年基金 (2016Q001)

利益冲突声明：本研究不存在研究者、伦理委员会成员、受试者监护人以及与公开研究成果有关的利益冲突。

作者贡献声明：王阳阳、安博然、贺文娟负责课题设计，资料分析，撰写论文；刘丛、黄薇、鲁素彩参与收集数据，修改论文；彭雪莹、陈岩负责数据整理，统计分析；孔立文、牛川、孟杰、侯英键负责拟定写作思路，指导撰写文章并最后定稿。

详细信息

作者简介:
王阳阳(1977—)，女，副教授，博士，主要从事肝脏疾病的研究

通信作者:
孟杰，cenci@163.com

中图分类号: R575
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- 被引次数: 0
出版历程
- 收稿日期: 2020-12-10
- 录用日期: 2021-02-01
- 出版日期: 2021-06-20

Protective effect of resolvin D1 pretreatment in a rat model of hepatic ischemia-reperfusion injury and its mechanism

1.
Department of Gastroenterology, Affiliated Hospital of Hebei University, Baoding, Hebei 071000, China
2.
School of Clinical Medicine, Hebei University, Baoding, Hebei 071000, China
3.
Department of Internal Medicine, Yixian People's Hospital, Baoding, Hebei 074200, China
4.
Department of Internal Medicine, Rongcheng Hospital of Traditional Chinese Medicine, Baoding, Hebei 071700, China
5.
Department of Internal Medicine, Baoding First Hospital of Traditional Chinese Medicine, Baoding, Hebei 071000, China

摘要

摘要: 目的探究消退素D1(RvD1)对肝缺血再灌注(IR)损伤大鼠模型的保护作用及与血红素氧合酶-1(HO-1)之间的关系。方法 Sprague-Dawley大鼠36只随机分为6组，分别为假手术(sham)+PBS组、sham+RvD1高剂量(10 μg/kg)组、IR+PBS组、IR+RvD1(2 μg/kg)低剂量组、IR+RvD1(5 μg/kg)中剂量组和IR+RvD1(10 μg/kg)高剂量组，每组6只。RvD1于缺血前1 h腹腔注射。生化仪测定ALT、AST水平，酶联免疫法检测血浆TNFα、IL-6、IL-8水平，HE染色观察肝组织学变化，Western Blot方法检测肝组织HO-1变化。计量资料多组间比较采用单因素方差分析，进一步两两比较采用LSD-t检验。结果与IR+PBS组相比，IR+RvD1中剂量组和IR+RvD1高剂量组大鼠ALT、AST水平以及炎症因子TNFα、IL-6、IL-8水平均明显降低(P值均＜0.05)，且中、高剂量两组间比较差异均无统计学意义(P值均＞0.05)。Western Blot结果显示IR+RvD1中剂量组和IR+RvD1高剂量组肝脏HO-1蛋白表达较IR+PBS组升高(P值均＜0.05)。HE染色观察肝组织学变化显示，与IR+PBS组相比，IR+RvD1中剂量组和IR+RvD1高剂量组细胞肿胀及肝索排列紊乱依然存在，但未见明显大片坏死区域。结论 RvD1可能通过增加肝脏HO-1表达，降低炎症因子(TNFα、IL-6、IL-8)和转氨酶(ALT、AST)水平，发挥对大鼠肝脏IR损伤的保护作用。
- 肝疾病 /
- 再灌注损伤 /
- 消退素D1 /
- 大鼠, Sprague-Dawley
Abstract: Objective To investigate the protective effect of resolvin D1 (RvD1) in a rat model of hepatic ischemia/reperfusion (IR) injury and its association with heme oxygenase-1 (HO-1). Methods A total of 36 Sprague-Dawley rats were randomly divided into sham-operation (sham)+phosphate-buffered saline (PBS) group, sham+high-dose RvD1 (10 μg/kg) group, IR+PBS group, IR+low-dose RvD1 (2 μg/kg) group, IR+middle-dose RvD1 (5 μg/kg) group, and IR+high-dose RvD1 (10 μg/kg) group, with 6 rats in each group. RvD1 were intraperitoneally injected at 1 hour before ischemia. A biochemical analyzer was used to measure the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST); ELISA was used to measure the plasma levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-8 (IL-8); HE staining was used to observe the histological changes of the liver; Western blot was used to measure the change in HO-1 in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. Results Compared with the IR+PBS group, the IR+middle-dose RvD1 group and the IR+high-dose RvD1 group had significant reductions in the levels of ALT, AST, and the inflammatory factors TNF-α, IL-6, and IL-8 (all P < 0.05), and there were no significant differences between the IR+middle-dose RvD1 group and the IR+high-dose RvD1 group (all P > 0.05). Western blot showed that compared with the IR+PBS group, the IR+middle-dose RvD1 group and the IR+high-dose RvD1 group had a significant increase in the protein expression of HO-1 in the liver (P < 0.05). HE staining showed that compared with the IR+PBS group, the IR+middle-dose RvD1 group and the IR+high-dose RvD1 group had cell swelling and disordered hepatic cords, without massive necrosis. Conclusion RvD1 exerts a protective effect against hepatic IR injury in rats by increasing the expression of HO-1 and reducing the levels of inflammatory factors (TNF-α, IL-6, and IL-8) and aminotransferases (ALT and AST).
- Liver Diseases /
- Reperfusion Injury /
- Resolvin D1 /
- Rats, Sprague-Dawley

HTML全文

图 1 各组大鼠肝脏组织学结果(HE染色，×100)

注：箭头所示为肝细胞坏死区域。

下载: 全尺寸图片幻灯片

图 2 Western Blot检测HO-1表达结果

注：与sham+PBS组比较，*P＜0.05；与IR+PBS组比较，#P＜0.05。

下载: 全尺寸图片幻灯片

表 1 实验动物分组

组别	过程
组别	缺血前1 h(腹腔注射)	缺血	缺血45 min后再灌注	再灌注8 h
sham+PBS组	PBS 1 ml	开关腹		取标本
sham+RvD1高剂量组	RvD1 10 μg/kg	开关腹		取标本
IR+PBS组	PBS 1 ml	钳夹缺血	再灌注，关腹	取标本
IR+RvD1低剂量组	RvD1 2 μg/kg	钳夹缺血	再灌注，关腹	取标本
IR+RvD1中剂量组	RvD1 5 μg/kg	钳夹缺血	再灌注，关腹	取标本
IR+RvD1高剂量组	RvD1 10 μg/kg	钳夹缺血	再灌注，关腹	取标本

下载: 导出CSV

表 2 各组大鼠血浆转氨酶水平比较

组别	动物数(只)	ALT(U/L)	AST(U/L)
sham+PBS组	6	56.88±11.65	58.45±12.11
sham+RvD1高剂量组	6	56.98±14.56	49.78±15.04
IR+PBS组	6	498.68±111.58¹⁾	606.35±150.06¹⁾
IR+RvD1低剂量组	6	578.23±89.50	610.92±77.30
IR+RvD1中剂量组	6	373.13±105.59²⁾	350.72±95.42²⁾
IR+RvD1高剂量组	6	348.17±86.52²⁾	369.72±94.25²⁾
F值		43.926	47.459
P值		＜0.001	＜0.001
注：与sham+PBS组比较，1)P＜0.05；与IR+PBS组比较，2)P＜0.05。

下载: 导出CSV

表 3 各组大鼠血浆炎症因子TNFα、IL-6、IL-8水平比较

组别	动物数(只)	IL-6(pg/ml)	IL-8(pg/ml)	TNFα(pg/ml)
sham+PBS组	6	26.32±7.05	120.83±19.17	37.30±6.59
sham+RvD1高剂量组	6	25.23±8.90	136.63±16.74	37.75±10.19
IR+PBS组	6	101.28±12.31¹⁾	697.88±152.13¹⁾	287.65±62.08¹⁾
IR+RvD1低剂量组	6	86.83±18.39	580.65±94.91	318.67±76.37
IR+RvD1中剂量组	6	42.47±12.94²⁾	399.13±75.17²⁾	83.55±12.34²⁾
IR+RvD1高剂量组	6	41.22±9.99²⁾	400.15±121.56²⁾	86.68±18.80²⁾
F值		42.276	36.249	56.206
P值		＜0.001	＜0.001	＜0.001
注：与sham+PBS组比较，1)P＜0.05；与IR+PBS组比较，2)P＜0.05。

下载: 导出CSV

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