Functional inhibition of TIM-4 in Kupffer cells alleviates hepatic ischemia-reperfusion injury in mice

Yan LIU; Hao WU; Jianping GONG; Xuhong LI

doi:10.16016/j.1000-5404.201806145

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Functional inhibition of TIM-4 in Kupffer cells alleviates hepatic ischemia-reperfusion injury in mice

Basic Medicine | 更新时间：2026-01-09

- Functional inhibition of TIM-4 in Kupffer cells alleviates hepatic ischemia-reperfusion injury in mice
- Journal of Third Military Medical University Vol. 41, Issue 1, Pages: 48-55(2019)
- 作者机构：
  
  1.611130 成都，成都市第五人民医院消化内科
  2.400010 重庆，重庆医科大学附属第二医院肝胆外科
  3.404000 重庆，重庆三峡中心医院百安分院外科
- 作者简介：
- 基金信息：
  
  the Key Science Research Project of Sichuan Provincial Health and Family Planning Committee(17ZD008);the Medical Science Research Project of Sichuan Province(S16007);the Medical Research Program of Chongqing Municipal Health and Family Planning Committee(2015MSXM123)
- DOI：10.16016/j.1000-5404.201806145
  CLC： R364.12;R392.32;R657.3
- Received：22 June 2018，
  
  Revised：2018-09-25，
  
  Published：15 January 2019
- 稿件说明：
移动端阅览
Yan LIU, Hao WU, Jianping GONG, et al. Functional inhibition of TIM-4 in Kupffer cells alleviates hepatic ischemia-reperfusion injury in mice[J]. Journal of Third Military Medical University, 2019, 41(1): 48-55.
DOI：

Yan LIU, Hao WU, Jianping GONG, et al. Functional inhibition of TIM-4 in Kupffer cells alleviates hepatic ischemia-reperfusion injury in mice[J]. Journal of Third Military Medical University, 2019, 41(1): 48-55. DOI： 10.16016/j.1000-5404.201806145.

摘要

目的

探讨阻断Kupffer细胞（Kupffer cells，KCs）的TIM-4蛋白功能对小鼠肝脏缺血再灌注损伤（hepatic ischemia reperfusion injury，HIRI）的影响。

方法

建立小鼠肝缺血再灌注损伤模型，术后Western blot检测KCs和树突状细胞（dendritic cells，DCs）TIM-4表达，免疫组化检测肝组织TIM-4表达；IR小鼠模型采用随机数字表法分为Sham组（PBS处理）、Ctr Ig组（TIM-4同型对照抗体处理）以及TIM-4 mAb组（TIM-4抗体处理），术后检测血清肝功、炎症指标、肝组织病理变化、肝细胞凋亡以及NF-κB通路相关蛋白的表达。

结果

KCs表达TIM-4蛋白随再灌注时间逐渐增加，然而DCs表达TIM-4不随时间变化；组化结果提示TIM-4主要表达于肝血窦巨噬细胞；与Ctr Ig组比较，TIM-4 mAb组血清肝功、促炎因子产物水平明显减低，而抗炎因子IL-10以及抗氧化应激产物CAT、SOD逐渐增高，且差异具有统计学意义（

0.05）；肝组织病理学结果示Ctr Ig组部分炎症细胞浸润，肝血窦轻度充血，TIM-4 mAb组无明显变化；TUNEL结果示，TIM-4 mAb组凋亡程度明显低于Ctr Ig组，Ctr Ig组和TIM-4 mAb组细胞早期凋亡程度分别为(25.56±1.26)%、(5.35±0.33)%，且两组差异具有统计学意义（

0.05）；阻断KCs TIM-4功能后，KCs TLR4以及下游p-IKKα、p-IκBα、p-p65蛋白表达水平逐渐减低；另外，TPCA-1（IKK-2阻断剂）进行预处理小鼠，进一步验证了阻断KCs TIM-4可通过抑制NF-κB信号通路减轻IRI程度。

结论

阻断KCs TIM-4的功能能够抑制NF-κB炎症通路的激活，改善IR炎症反应以及肝细胞凋亡，从而对HIRI起到一定的保护作用。

Abstract

Objective

To investigate the effects of functional inhibition of TIM-4 in Kupffer cells(KCs) in protecting against hepatic ischemia-reperfusion injury in mice and explore the mechanism.

Methods

We determined the expression of TIM-4 in KCs and dendritic cells(DCs) in a mouse model of hepatic ischemia-reperfusion injury using Western blotting and immunohistochemistry. The mouse models were treated with PBS(sham group)

a monoclonal antibody of TIM-4(TIM-4 mAb group) or a control antibody(Ctr Ig group)

and the liver functions

inflammatory factors and products of oxidative stress were assayed. HE staining was used to observe the pathological changes in the liver tissue

and TUNEL and flow cytometry were used to detect apoptosis of the hepatocytes. Western blotting was used to detect the expression of nuclear factor-κB(NF-κB) pathway-related proteins.

Results

In the mouse model of hepatic ischemia-reperfusion injury

the expression of TIM-4 protein in KCs was increased progressively as the reperfusion time extended

while TIM-4 expression in the DCs showed no significant variations with time. Immunohistochemical staining identified TIM-4 expression mainly in the hepatic sinusoidal macrophages; The serum levels of hepatic function-related factors

pro-inflammatory cytokines in the hepatic homogenate

and oxidative stress products were significantly reduced

while anti-inflammatory cytokine(IL-10) and anti-oxidative stress products(CAT and SOD) were increased significantly in TIM-4 mAb group as compared with Ctr Ig group(

0.05). Histopathological examination of the liver tissues revealed inflammatory cell infiltration and mild congestion in the hepatic sinusoids in Ctr Ig group

but no obvious pathological changes in TIM-4 mAb group. The apoptotic index was significantly lower in TIM-4 mAb grou

p than in Ctr Ig group; the early apoptosis rates in the 2 groups were(5.35±0.33)% and(25.56±1.26)%

respectively

showing a significant difference between them(

0.05). Functional blocking of TIM-4 in the KCs lowered the expression levels of TLR4 and the downstream p-IKKα

p-IκBα

p-p65 proteins. Pre-treatment of the mice with TPCA-1

an IKK-2 blocker

further validated that blocking TIM-4 in the KCs inhibited NF-κB signaling pathway and improved hepatic ischemia-reperfusion injury.

Conclusion

Functional inhibition of TIM-4 in the KCs can inhibit the activation of NF-κB inflammatory pathway and reduce the inflammatory response and hepatocyte apoptosis to protect against hepatic ischemia-reperfusion injury in mice.

关键词

Keywords

references

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