DIDS alleviates hepatic ischemia-reperfusion injury by inhibiting the opening of type 2 chloride channel in rats

Yong DENG; Tengqian TANG; Qingyi ZHANG; Leida ZHANG

doi:10.16016/j.1000-5404.202007131

您当前的位置：

首页 >

文章列表页 >

DIDS alleviates hepatic ischemia-reperfusion injury by inhibiting the opening of type 2 chloride channel in rats

Basic Medicine | 更新时间：2026-01-09

- DIDS alleviates hepatic ischemia-reperfusion injury by inhibiting the opening of type 2 chloride channel in rats
- Journal of Third Military Medical University Vol. 42, Issue 24, Pages: 2375-2381(2020)
- 作者机构：
  
  400038 重庆，陆军军医大学(第三军医大学)第一附属医院肝胆外科
- 作者简介：
- 基金信息：
- DOI：10.16016/j.1000-5404.202007131
  CLC： R364.12;R657.3;R966
- Received：15 July 2020，
  
  Revised：2020-09-11，
  
  Published：30 December 2020
- 稿件说明：
移动端阅览
Yong DENG, Tengqian TANG, Qingyi ZHANG, et al. DIDS alleviates hepatic ischemia-reperfusion injury by inhibiting the opening of type 2 chloride channel in rats[J]. Journal of Third Military Medical University, 2020, 42(24): 2375-2381.
DOI：

Yong DENG, Tengqian TANG, Qingyi ZHANG, et al. DIDS alleviates hepatic ischemia-reperfusion injury by inhibiting the opening of type 2 chloride channel in rats[J]. Journal of Third Military Medical University, 2020, 42(24): 2375-2381. DOI： 10.16016/j.1000-5404.202007131.

摘要

目的

探讨氯离子通道2型(type 2 chloride channel

ClC2)及其阻断剂4

4-二异硫氰基芪-2

2’-二磺酸(4

4’-diisothiocyanostilbene-2

2’-disulfonic acid

DIDS)在肝缺血再灌注损伤中的作用。

方法

成年SD大鼠以经典法建立肝缺血再灌注模型。采用q-PCR技术对缺血再灌注后肝组织中ClC2的表达进行检测，采用肝功能血生化分析、活性氧浓度检测、ELISA及流式细胞技术观察缺血再灌注及应用DIDS后对肝功能、氧自由基水平、炎性细胞因子及细胞周期等方面的影响。

结果

缺血再灌注后肝组织中ClC2 mRNA表达显著升高(

0.05)，并伴随有活性氧浓度升高(

0.05)、谷丙及谷草转氨酶升高(

0.05)和炎性细胞因子浓度升高。术后即时使用DIDS组相比缺血再灌注组在ClC2表达、转氨酶浓度、活性氧水平及炎性因子表达方面均有所逆转(

0.05)，术后8 h应用DIDS组相比缺血再灌注组在ClC2表达、转氨酶浓度及iNOS(一氧化氮合酶)表达方面有显著差异(

0.05)，而预用DIDS 1 h组相比缺血再灌注组未观察到统计学差异。

结论

DIDS可以对肝缺血再灌注损伤引起的肝实质损害、炎性反应及氧化应激起保护作用。

Abstract

Objective

To investigate the roles of type 2 chloride channel (ClC2) and its blocker

4'-diisothiocyanostilbene-2

2'-disulfonic acid (DIDS)

in rat model of hepatic ischemia-reperfusion (I/R) injury.

Methods

A total of 75 health adult SD rats were randomly divided into sham operation group

hepatic I/R injury group

I/R+immediate treatment of DIDS group

I/R+8 h later treatment of DIDS group

and 1 h pretreatment of DIDS+I/R group

with 15 animals in each group. The rat model of hepatic I/R injury was established by classical method

and DIDS was given by intraperitoneal injection at a dose of 50 mg/kg at corresponding time points. The expression of ClC2 in the liver tissues was detected with q-PCR. Then blood biochemical analysis

detection of reactive oxygen concentration

ELISA and flow cytometry were employed respectively to analyze and compare the liver function

level of reactive oxygen species (ROS)

contents of inflammatory cytokines and cell cycle of hepatic cells among the different groups.

Results

The rats with hepatic I/R injury had significantly enhanced expression of ClC2 mRNA (

0.05)

increased production of ROS (

0.05)

elevated contents of inflammatory cytokines in liver tissues

and raised serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) (

0.05). However

all above phenomena were reversed in the rats from the I/R+immediate treatment of DIDS group when compared with the model group (

0.05). Significant differences were also observed in the ClC2 expression

transaminase concentrations

and inducible nitric oxide synthase (iNOS) level between the model group and the group using DIDS 8 h after operation (

0.05). But no statistical difference was found in comparison with the group undergoing pretreatment of DIDS 1 h before operation.

Conclusion

DIDS has a protective effect on liver parenchymal damage

inflammatory reaction and oxidative stress induced by hepatic I/R injury.

关键词

Keywords

references

CHEN S Y, ZHANG H P, LI J, et al . Tripartite motif-containing 27 attenuates liver ischemia/reperfusion injury by suppressing TAK1 via TAB2/3 degradation[J ] . Hepatology, 2020, (4): 1-38. DOI: 10.1002/hep.31295.

ZHANG T, GUO J, GU J, et al . Protective role of mTOR in liver ischemia/reperfusion injury: involvement of inflammation and autophagy[J ] .Oxid Med Cell Longev, 2019, 13: 1-17. DOI: 10.1155/2019/7861290 .

ZHAO B X, QUAN H Y, MA T, et al . 4, 4'-diisothiocyanostilbene-2, 2'-disulfonic acid (DIDS) ameliorates ischemia-hypoxia-induced white matter damage in neonatal rats through inhibition of the voltage-gated chloride channel ClC-2[J ] . Int J Mol Sci, 2015, 16(5): 10457-10469. DOI: 10.3390/ijms160510457.

WANG H, XU M, KONG Q, et al . Research and progress on ClC-2[J ] . Mol Med Rep, 2017, 16(1): 11-22. DOI: info:doi/10.3892/mmr.2017.6600.

HOSSEINZADEH Z, BHAVSAR S K, LANG F. Downregulation of ClC-2 by JAK2[J]. Cell Physiol Biochem, 2012, 29(5/6): 737-742. DOI: 10.1159/000178560.

VIJ N, ZEITLIN P L. Regulation of the ClC-2 lung epithelial chloride channel by glycosylation of SP1[J]. Am J Respir Cell Mol Biol, 2006, 34(6): 754-759. DOI: 10.1165/rcmb.2005-0442OC.

CUPPOLETTI J, CHAKRABARTI J, TEWARI K P, et al . Differentiation between human ClC-2 and CFTR Cl-channels with pharmacological agents[J ] . Am J Physiol Cell Physiol, 2014, 307(5): C479-C492. DOI: 10.1152/ajpcell.00077.2014.

WANG X M, CAO Y N, SHEN M Z, et al . DIDS reduces ischemia/reperfusion-induced myocardial injury in rats[J ] . Cell Physiol Biochem, 2015, 35(2): 676-688. DOI: 10.1159/000369728.

BOLLMANN E, SCHREIBER J A, RITTER N, et al . 4, 4'-diisothiocyanato-2, 2'-stilbenedisulfonic acid (DIDS) modulates the activity of KCNQ1/KCNE1 channels by an interaction with the central pore region[J ] . Cell Physiol Biochem, 2020, 54(2): 321-332. DOI: 10.33594/000000222.

XIE S Q, WANG K, YU W J, et al . DIDS blocks a chloride-dependent current that is mediated by the 2B protein of Enterovirus 71[J ] . Cell Res, 2011, 21(8): 1271-1275. DOI: 10.1038/cr.2011.112.

HEUSCH G. Remote ischemic conditioning: the enigmatic transfer of protection[J]. Cardiovasc Res, 2017, 113(1): 1-2. DOI: 10.1093/cvr/cvw240.

LAGOSTENA L, ZIFARELLI G, PICOLLO A. New insights into the mechanism of NO3- selectivity in the human kidney chloride channelClC-ka and the CLC protein family[J]. J Am Soc Nephrol, 2019, 30(2): 293-302. DOI: 10.1681/ASN.2018060593.

SCHEWE J, SEIDEL E, FORSLUND S, et al . Elevated aldosterone and blood pressure in a mouse model of familial hyperaldosteronism with ClC-2 mutation[J ] . Nat Commun, 2019, 10(1): 5155. DOI: 10.1038/s41467-019-13033-4.

QIU L Y, CHEN H P, YAN Y F, et al .Sasanquasaponin promotes cellular chloride efflux and elicits cardioprotection via the PKCε pathway[J ] . Mol Med Rep, 2016, 13(4): 3597-3603. DOI: 10.3892/mmr.2016.4984.

NOZIK-GRAYCK E, PIANTADOSI C A, VAN ADELSBERG J, et al . Protection of perfused lung from oxidant injury by inhibitors of anion exchange[J ] . Am J Physiol Lung Cell Mol Physiol, 1997, 273(2): L296-L304. DOI: 10.1152/ajplung.1997.273.2.l296.

ZHU B M, MIYAMOTO S, NAGASAWA Y, et al . Does Cl-/HCO3- exchange play an important role in reperfusion arrhythmias in rats?[J ] . Eur J Pharmacol, 2003, 460(1): 43-50. DOI: 10.1016/s0014-2999(02)02928-x.

XUE J, ZHOU D, YAO H, et al . Role of transporters and Ion channels in neuronal injury under hypoxia[J ] . Am J Physiol Regul Integr Comp Physiol, 2008, 294(2): R451-R457. DOI: 10.1152/ajpregu.00528.2007.

PALMADA M, DIETER M, BOEHMER C, et al . Serum and glucocorticoid inducible kinases functionally regulate ClC-2 channels[J ] .Biochem Biophys Res Commun, 2004, 321(4): 1001-1006. DOI: 10.1016/j.bbrc.2004.07.064.

WARSI J, ELVIRA B, HOSSEINZADEH Z, et al . Downregulation of chloride channel ClC-2 by Janus kinase 3[J ] . J Membr Biol, 2014, 247(5): 387-393. DOI: 10.1007/s00232-014-9645-0.

HOLMES K W, HALES R, CHU S J, et al . Modulation of Sp1 and Sp3 in lung epithelial cells regulates ClC-2 chloride channel expression[J ] . Am J Respir Cell Mol Biol, 2003, 29(4): 499-505. DOI: 10.1165/rcmb.2003-0030oc.

KHANDOGA A, BIBERTHALER P, ENDERS G, et al . P-selectin mediates platelet-endothelial cell interactions and reperfusion injury in the mouse liver in vivo[J ] . Shock, 2002, 18(6): 529-535. DOI: 10.1097/00024382-200212000-00008.

RENTSCH M, POST S, PALMA P, et al . Anti-ICAM-1 blockade reducespostsinusoidal WBC adherence following cold ischemia and reperfusion, but does not improve early graft function in rat liver transplantation[J ] . J Hepatol, 2000, 32(5): 821-828. DOI: 10.1016/s0168-8278(00)80252-4.

HARRIS I S, DENICOLA G M. The complex interplay between antioxidants and ROS in cancer[J]. Trends Cell Biol, 2020, 30(6): 440-451. DOI: 10.1016/j.tcb.2020.03.002.

ZHAO G Y, FU C, WANG L, et al . Down-regulation of nuclear HMGB1 reduces ischemia-induced HMGB1 translocation and release and protects against liver ischemia-reperfusion injury[J ] . Sci Rep, 2017, 7: 46272. DOI: 10.1038/srep46272.

XIANG B, ZHONG P, FANG L, et al . miR-183 inhibits microglia activation and expression of inflammatory factors in rats with cerebral ischemia reperfusion via NF-κB signaling pathway[J ] . Exp Ther Med, 2019, 18(4): 2540-2546. DOI: 10.3892/etm.2019.7827.

NING X J, YAN X, WANG Y F, et al . Parkin deficiency elevates hepatic ischemia/reperfusion injury accompanying decreased mitochondrial autophagy, increased apoptosis, impaired DNA damage repair and altered cell cycle distribution[J ] . Mol Med Rep, 2018, 18(6): 5663-5668. DOI: 10.3892/mmr.2018.9606.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Establishment of a mouse model of skin-dominant chronic graft-versus-host disease

Expression profile of AQP9 in liver of rats with type 2 diabetes mellitus

Myocardial protective effect of propofol postconditioning in renal hypertensive rats

Effect of dexmedetomidine on spinal cord ischemia-reperfusion injury in rats

Effect of rapamycin on mitochondrial DNA damage in rats after liver ischemia-reperfusion injury and repair mechanisms

Related Author

Weihao WANG

Shijie YANG

Yuxuan DU

Xiaoqi WANG

Tao HONG

Yundi HE

Rui WANG

Chao MA

Related Institution

State Key Laboratory of Trauma, Burns and Combined Injury, Medical Center of Hematology, PLA Center for Hematology, Second Affiliated Hospital, Army Medical University (Third Military Medical University)

Department of Clinical Laboratory, the Second Affiliated Hospital of Zunyi Medical University

Department of Cell Biology, College of Basic Medical Sciences, Army Medical University (Third Military Medical University)

Neuroscience Center, Chongqing Medical University

Seven-year Training Group of Grade 2013, First Clinical College of Chongqing Medical University

⁰