RANKL在小鼠肝纤维化形成中抑制肝星状细胞的活化

周丽佳; 钟立; 薛倩; 邓亮; 黄璐

doi:10.16016/j.2097-0927.202205095

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RANKL在小鼠肝纤维化形成中抑制肝星状细胞的活化

基础医学 | 更新时间：2026-01-09

- RANKL在小鼠肝纤维化形成中抑制肝星状细胞的活化
- RANKL inhibits activation of hepatic stellate cells during hepatic fibrosis in mice
- 陆军军医大学学报 2023年45卷第2期页码：139-145
- 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：
  
  [黄璐, E-mail：huanglu_86oct@163.com HUAGN Lu, E-mail: huanglu_86oct@163.com
- 基金信息：
  
  重庆市自然科学基金面上项目(cstc2021jcyj-msxmX0215)
- DOI：10.16016/j.2097-0927.202205095
  中图分类号： R322.47;R341;R363.21
- 收稿：2022-05-17，
  
  修回：2022-09-18，
  
  纸质出版：2023-01-30
- 稿件说明：
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周丽佳, 钟立, 薛倩, 等. RANKL在小鼠肝纤维化形成中抑制肝星状细胞的活化[J]. 陆军军医大学学报, 2023,45(2):139-145.

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.
周丽佳, 钟立, 薛倩, 等. RANKL在小鼠肝纤维化形成中抑制肝星状细胞的活化[J]. 陆军军医大学学报, 2023,45(2):139-145. DOI： 10.16016/j.2097-0927.202205095.

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

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

将26只6~8周龄雄性C57BL/6鼠随机分为(

=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

)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

造模组小鼠肝组织中RANKL的mRNA和蛋白表达水平显著增高(

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(

0.001)

且在CCL

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(

0.05或

0.01或

0.001)

且有剂量依赖性。

结论

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

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

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

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

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

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

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 (

0.001)

while CCL

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 (

0.05

0.01 or

0.001).

Conclusion

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

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