RANKL inhibits activation of hepatic stellate cells during hepatic fibrosis in mice

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RANKL inhibits activation of hepatic stellate cells during hepatic fibrosis in mice

Basic Medicine | 更新时间：2026-01-09

- RANKL inhibits activation of hepatic stellate cells during hepatic fibrosis in mice
- Journal of Army Medical University Vol. 45, Issue 2, Pages: 139-145(2023)
- 作者机构：
  
  1.400014 重庆，重庆医科大学附属儿童医院儿科研究所，国家儿童健康与疾病临床医学研究中心，儿童发育疾病研究教育部重点实验室，儿童发育重大疾病国家国际科技合作基地，儿童感染免疫重庆市重点实验室
  2.400010 重庆，重庆医科大学附属第二医院消化内科
  3.400016 重庆，重庆医科大学附属第一医院消化内科
- 作者简介：
- 基金信息：
  
  the General Program of Natural Science Foundation of Chongqing(cstc2021jcyj-msxmX0215)
- DOI：10.16016/j.2097-0927.202205095
  CLC： R322.47;R341;R363.21
- Received：17 May 2022，
  
  Revised：2022-09-18，
  
  Published：30 January 2023
- 稿件说明：
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Lijia ZHOU, Li ZHONG, Qian XUE, et al. RANKL inhibits activation of hepatic stellate cells during hepatic fibrosis in mice[J]. Journal of Army Medical University, 2023, 45(2): 139-145.
DOI：

Lijia ZHOU, Li ZHONG, Qian XUE, et al. RANKL inhibits activation of hepatic stellate cells during hepatic fibrosis in mice[J]. Journal of Army Medical University, 2023, 45(2): 139-145. DOI： 10.16016/j.2097-0927.202205095.

摘要

目的

2

探讨核因子-κB受体活化因子配体(receptor activator of nuclear factor kappa-B ligand

RANKL)在小鼠肝星状细胞(hepatic stellate cells

HSCs)活化及肝纤维化进程中的生物学作用。

方法

2

将26只6~8周龄雄性C57BL/6鼠随机分为(

n

=13): 肝纤维化造模组[按照5 μL/g腹腔

注射体积分数10%的四氯化碳(carbon tetracholoride

CCL

4

)油剂溶液

2次/周诱导小鼠肝纤维化

]

和对照组(同时注射等体积的无菌油剂)。6周后通过RT-qPCR、IHC和Western blot等方法检测肝组织、原代肝细胞(hepatocyte

HC)和HSCs中的RANKL

α-平滑肌动蛋白(α-SMA)和Ⅰ型胶原(Collagen-Ⅰ)的mRNA及蛋白表达水平。用不同浓度外源性重组RANKL细胞因子培养原代HSCs

采用RT-qPCR和IF分别检测各组RANKL、α-SMA和Collagen-ⅠmRNA及蛋白表达水平。

结果

2

造模组小鼠肝组织中RANKL的mRNA和蛋白表达水平显著增高(

P

<

0.05)

且蛋白表达呈弥散分布; 对照组小鼠体内

RT-qPCR和Western blot结果均显示HC的RANKL mRNA和蛋白表达水平高于在HSCs中的表达水平(

P

<

0.001)

且在CCL

4

诱导小鼠肝纤维化后

HC的RANKL表达水平进一步升高

而HSCs的RANKL表达水平出现下降; 在体外刺激原代HSCs活化时加入不同浓度的RANKL重组细胞因子后

与对照组相比

加入重组RANKL后的HSCs在活化过程中RANKL、α-SMA和Collagen-ⅠmRNA和蛋白表达水平均降低(

P

<

0.05或

P

<

0.01或

P

<

0.001)

且有剂量依赖性。

结论

2

在小鼠肝纤维化的进程中

HC表达大量RANKL

而RANKL可抑制HSCs的活化。

Abstract

Objective

2

To explore the biological role of receptor activator of nuclear factor kappa-B ligand (RANKL) in the activation of hepatic stellate cells (HSCs) and the process of hepatic fibrosis.

Methods

2

Twenty-six male C57BL/6 mice (6~8 weeks old) were randomly assigned to control group and liver fibrosis group (

n

=13).The mice in the latter group were injected intraperitoneally with 5 μL/g 10% carbon tetracholoride (CCL

4

twice/week) to induce liver fibrosis

while those of the former group were given same volume of sterile oil solution.In 6 weeks later

the expression of RANKL

α-SMA and Collagen-Ⅰ at mRNA and protein levels in liver tissue

primary hepatocytes (HCs) and HSCs was detected by quantitative real-time PCR (RT-qPCR)

immunohistochemical assay and Western blotting

respectively.After primary HSCs were treated with different concentrations of exogenous recombinant RANKL

the mRNA and protein levels of above molecules were measured with RT-qPCR and immunofluorescence assay.

Results

2

The mRNA and protein levels of RANKL were significantly increased in the liver tissue from the model group than the control group (

P

<

0.05)

and the protein was in a diffuse distribution.In the control mice

the mRNA and protein levels of RANKL was obviously higher in HCs than HSCs (

P

<

0.001)

while CCL

4

treatment induced liver fibrosis resulted in the expression level of RANKL was further increased in HCs

while decreased in HSCs.After different concentrations of recombinant RANKL were added to stimulate the activation of primary HSCs

the expression of RANKL

α-SMA and Collagen-Ⅰat mRNA and protein levels in HSCs activated by recombinant RANKL was decreased in a dose-dependent manner (

P

<

0.05

P

<

0.01 or

P

<

0.001).

Conclusion

2

In the process of liver fibrosis in mice

HCs express a large amount of RANKL

and RANKL can inhibit the activation of HSCs.

关键词

Keywords

references

DUVAL F, MORENO-CUEVAS J E, GONZÁLEZ-GARZA M T, et al . Liver fibrosis and mechanisms of the protective action of medicinal plants targeting inflammation and the immune response[J ] . Int J Inflam, 2015, 2015: 943497. DOI: 10.1155/2015/943497.

VAN DIJK F, HAZELHOFF C M, POST E, et al . Design of a gene panel to expose the versatile role of hepatic stellate cells in human liver fibrosis[J ] . Pharmaceutics, 2020, 12(3): 278. DOI: 10.3390/pharmaceutics12030278.

TSUCHIDA T, FRIEDMAN S L. Mechanisms of hepatic stellate cell activation[J]. Nat Rev Gastroenterol Hepatol, 2017, 14(7): 397-411. DOI: 10.1038/nrgastro.2017.38.

KOYAMA Y, BRENNER D A. Liver inflammation and fibrosis[J]. J Clin Invest, 2017, 127(1): 55-64. DOI: 10.1172/JCI88881.

WONG B R, JOSIEN R, LEE S Y, et al . TRANCE (tumor necrosis factor[TNF ] -related activation-induced cytokine), a new TNF family member predominantly expressed in T cells, is a dendritic cell-specific survival factor[J ] . J Exp Med, 1997, 186(12): 2075-2080. DOI: 10.1084/jem.186.12.2075.

WALSH M C, CHOI Y. Biology of the RANKL-RANK-OPG system in immunity, bone, and beyond[J]. Front Immunol, 2014, 5: 511. DOI: 10.3389/fimmu.2014.00511.

LACEY D L, TIMMS E, TAN H L, et al . Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation[J ] . Cell, 1998, 93(2): 165-176. DOI: 10.1016/s0092-8674(00)81569-x.

VIDELA L A, TAPIA G, RODRIGO R, et al . Liver NF-κB and AP-1 DNA binding in obese patients[J ] . Obesity, 2009, 17(5): 973-979. DOI: 10.1038/oby.2008.601.

TAT S K, PELLETIER J P, VELASCO C R, et al . New perspective in osteoarthritis: the OPG and RANKL system as a potential therapeutic target?[J ] . Keio J Med, 2009, 58(1): 29-40. DOI: 10.2302/kjm.58.29.

LLEO A, BIAN Z, ZHANG H, et al . Quantitation of the rank-rankl axis in primary biliary cholangitis[J ] . PLoS One, 2016, 11(9): e0159612. DOI: 10.1371/journal.pone.0159612.

TAKENO A, YAMAMOTO M, NOTSU M, et al . Administration of anti-receptor activator of nuclear factor-kappa B ligand (RANKL) antibody for the treatment of osteoporosis was associated with amelioration of hepatitis in a female patient with growth hormone deficiency: a case report[J ] . BMC Endocr Disord, 2016, 16(1): 66. DOI: 10.1186/s12902-016-0148-0.

SAKAI N, VAN SWERINGEN H L, SCHUSTER R, et al . Receptor activator of nuclear factor-κB ligand (RANKL) protects against hepatic ischemia/reperfusion injury in mice[J ] . Hepatology, 2012, 55(3): 888-897. DOI: 10.1002/hep.24756.

FAOUZI S, LE BAIL B, NEAUD V, et al . Myofibroblasts are responsible for collagen synthesis in the stroma of human hepatocellular carcinoma: an in vivo and in vitro study[J ] . J Hepatol, 1999, 30(2): 275-284. DOI: 10.1016/s0168-8278(99)80074-9..

ZHONG L, YUAN J H, HUANG L, et al . RANKL is involved in Runx2-triggered hepatic infiltration of macrophages in mice with NAFLD induced by a high-fat diet[J ] . Biomed Res Int, 2020, 2020: 6953421. DOI: 10.1155/2020/6953421.

LI Y J, CHEN Y Y, HUANG H Y, et al . Autophagy mediated by endoplasmic Reticulum stress enhances the caffeine-induced apoptosis of hepatic stellate cells[J ] . Int J Mol Med, 2017, 40(5): 1405-1414. DOI: 10.3892/ijmm.2017.3145.

REN J J, HUANG T J, ZHANG Q Q, et al . Insulin-like growth factor binding protein related protein 1 knockdown attenuates hepatic fibrosis via the regulation of MMPs/TIMPs in mice[J ] . Hepatobil Pancreat Dis Int, 2019, 18(1): 38-47. DOI: 10.1016/j.hbpd.2018.08.008.

SEKI E, DE MINICIS S, ÖSTERREICHER C H, et al . TLR4 enhances TGF-β signaling and hepatic fibrosis[J ] . Nat Med, 2007, 13(11): 1324-1332. DOI: 10.1038/nm1663.

HE X, PU G B, TANG R, et al . Activation of nuclear factor kappa B in the hepatic stellate cells of mice with schistosomiasis Japonica[J ] . PLoS One, 2014, 9(8): e104323. DOI: 10.1371/journal.pone.0104323.

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Related Author

Zhen HU

Li ZHONG

Liang DENG

Chang LIU

TANG Yin

ZHANG Jie

MAO Cui

XIONG Tao

Related Institution

Department of Laboratory Medicine, the First Affiliated Hospital of Chongqing Medical University

Department of Gastroenterology, the Second Affiliated Hospital of Chongqing Medical University

Department of Gastroenterology, the First Affiliated Hospital of Chongqing Medical University

Department of Critical Care Medicine, the First Affiliated Hospital of Chongqing Medical University

Department of Hepatobiliary Surgery， First Affiliated Hospital（Southwest Hospital）， Army Medical University （Third Military Medical University）

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