[1]徐雪松,王孟皓,白赫,等.牛磺熊去氧胆酸抑制Kupffer细胞减轻大鼠肝移植后缺血再灌注损伤[J].第三军医大学学报,2018,40(18):1653-1662.
 XU Xuesong,WANG Menghao,BAI He,et al.Tauroursodeoxycholic acid alleviates ischemia-reperfusion injury in liver graft following transplantation by suppressing Kupffer cell function in rats[J].J Third Mil Med Univ,2018,40(18):1653-1662.
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牛磺熊去氧胆酸抑制Kupffer细胞减轻大鼠肝移植后缺血再灌注损伤(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第18期
页码:
1653-1662
栏目:
基础医学
出版日期:
2018-09-30

文章信息/Info

Title:
Tauroursodeoxycholic acid alleviates ischemia-reperfusion injury in liver graft following transplantation by suppressing Kupffer cell function in rats
作者:
徐雪松王孟皓白赫龚建平
重庆医科大学附属第二医院肝胆外科
Author(s):
XU Xuesong WANG Menghao BAI He GONG Jianping

Department of Hepatobiliary Surgery, Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, China

关键词:
牛磺熊去氧胆酸内质网应激肝脏缺血再灌注损伤Kupffer细胞
Keywords:
tauroursodeoxycholic acid endoplasmic reticulum stress hepatic ischemia-reperfusion injury Kupffer cells
分类号:
R284.1;R617;R657.3
文献标志码:
A
摘要:

目的    探讨牛磺熊去氧胆酸(tauroursodeoxycholic acid,TUDCA)减轻大鼠肝移植后肝脏缺血再灌注损伤的机制。方法    分离雄性SD大鼠Kupffer细胞并培养,按不同浓度梯度给予TUDCA体外处理24 h,收集细胞上清液并提取蛋白和mRNA。共分为4组:①非活化组;②活化+0 μmol/L组;③活化+50 μmol/L组;④活化+100 μmol/L组。检测炎症因子、IRE1α/TRAF2/NFκB通路活性、P65入核程度。雄性SD大鼠于术前3 d经腹腔注射TUDCA 400 mg·kg-1·d-1或者等量生理盐水后,建立大鼠肝移植模型,分为3组:①假手术组;②缺血再灌注组(IR组);③IR+TUDCA组。各组大鼠分别于再灌注3、6、24 h后收取肝组织标本以及血液标本,并提取Kupffer细胞中蛋白。检测肝脏病理改变、肝功能、肝细胞凋亡程度、肝脏炎症因子、Kupffer细胞中IRE1α/TRAF2/NFκB通路活性。结果    TUDCA处理活化的Kupffer细胞后,IL-1β、IL-6、TNF-α分泌减少(P<0.05),并且IRE1α、TRAF2、p-IKKα、p-IκB、p-P65表达降低(P<005),P65细胞核相对表达程度降低。TUDCA干预大鼠肝移植模型后,肝组织中气球样变性、点灶坏死,汇管区大量中性粒细胞、淋巴细胞浸润,肝血窦变窄等病理改变显著减轻,IR+TUDCA组Suzuk’s评分(2.95±0.15)较IR组(4.78±0.36)明显降低,肝功能标志物水平下降(P<0.05),肝实质细胞凋亡数量减少(P<0.05),IR+TUDCA组凋亡指数(29.1±12.3)较IR组(56.3±17.5)明显降低(P<0.05),并且IL-1β、IL-6、TNF-α分泌减少(P<0.05),Kupffer细胞中IRE1α、TRAF2、p-IKKα、p-IκB、p-P65表达降低(P<0.05)。结论     牛磺熊去氧胆酸能够减轻大鼠肝移植后肝脏缺血再灌注损伤程度,其机制可能是下调IRE1α/TRAF2/NF-κB通路活性抑制Kupffer细胞功能。

Abstract:

Objective     To investigate the protective effect of tauroursodeoxycholic acid (TUDCA) against ischemiareperfusion (IR) injury in the liver graft after transplantation and explore the possible mechanism. Methods    Cultured Kupffer cells from male SD rats were activated by stimulation with lipopolysaccharide (LPS) followed by treatment with TUDCA (50 or 100 μmol/L) or PBS for 24 h, and the inflammatory factors and the activity of the IRE1α/TRAF2/NF-κB pathway activity in the supernatant were detected. Male SD rats were divided into sham-operation group, IR group, and IR+TUDCA group (n=15), and in the latter 2 groups, the rats were subjected to daily intraperitoneal injection of normal saline and TUDCA (400 mg/kg) for 3 consecutive days, respectively, before orthotopic liver transplantation was performed. Blood and liver graft samples were collected from the recipient rats at 3, 6, and 24 h after the operation for assessing the graft pathologies, liver function, hepatocyte apoptosis and the proinflammatory factors, and Kupffer cells were isolated to detect the activity of the IRE1α/TRAF2/NF-κB pathway. Results    Treatment with TUDCA significantly decreased the secretion of IL-1β, IL-6 and TNF-α (P<0.05), lowered the expression levels of IRE1α, TRAF2, p-IKKα, p-IκB, and p-P65 (P<0.05), and reduced intranuclear expression of P65 in LPS-activated Kupffer cells. In the rat models of orthotopic liver transplantation, compared with saline treatment, TUDCA treatment obviously alleviated IR-induced pathologies (including ballooning degeneration, spotted and focal necrosis, massive accumalation of neutrophils in portal area, lymphocyte infiltration, and narrowing of hepatic sinuses) in the liver graft, significantly lowered the Suzuk’s score from 4.78±0.36 to 2.95±0.15 (P<0.05), and reduced the levels of alanine aminotransferase and aspartate transaminase (P<0.05) and hepatocyte apoptosis (P<0.05). TUDCA treatment significantly lowered the apoptotic index of the graft from 56.3±17.5 (in salinetreated IR rats) to 29.1±12.3 (P<0.05), and obviously suppressed the secretion of IL-1β, IL-6, and TNF-α (P<0.05) and the expression  levels of IRE1α, TRAF2, p-IKKα, p-IκB, and p-P65 (P<0.05) in the Kupffer cells from the recipient rats. Conclusion    TUDCA can effectively alleviate IR injury in the liver graft possibly by down-regulating the activity of IRE1α/TRAF2/NF-κB pathway to suppress the function of the Kupffer cells.

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更新日期/Last Update: 2018-09-28