High-salt diet promotes the progression of fibrosis in metabolic dysfunction-associated steatohepatitis <italic style="font-style: italic">via</italic> activating the PI3K/Akt/mTOR signaling pathway

TANG Yin; ZHANG Jie; MAO Cui; XIONG Tao; XIE Chuanming; ZHANG Leida

doi:10.16016/j.2097-0927.202511049

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High-salt diet promotes the progression of fibrosis in metabolic dysfunction-associated steatohepatitis via activating the PI3K/Akt/mTOR signaling pathway

Basic Medicine | 更新时间：2025-12-30

- High-salt diet promotes the progression of fibrosis in metabolic dysfunction-associated steatohepatitis via activating the PI3K/Akt/mTOR signaling pathway
- Journal of Army Medical University Vol. 47, Issue 24, Pages: 3009-3021(2025)
- 作者机构：
  
  陆军军医大学（第三军医大学）第一附属医院（西南医院）全军肝胆外科研究所，重庆
- 作者简介：
  
  XIE Chuanming，E-mail： chuanming506@126.com
  ZHANG Leida；E-mail： zld666@aliyun.com
- 基金信息：
  
  the National Natural Science Foundation for Young Scholars of China(82203780;82504065)
- DOI：10.16016/j.2097-0927.202511049
  CLC： R363.21;R575.5;R589.4
- Received：10 November 2025，
  
  Revised：2025-11-28，
  
  Published：30 December 2025
- 稿件说明：
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唐银，张捷，毛毳，等.高盐饮食通过PI3K/AKT/mTOR通路激活促进代谢功能障碍相关脂肪性肝炎纤维化进展［J］.陆军军医大学学报，2025，47（24）：3009-3021.DOI：10.16016/j.2097-0927.202511049.

TANG Y，ZHANG J，MAO C，et al.High-salt diet promotes the progression of fibrosis in metabolic dysfunction-associated steatohepatitis via activating the PI3K/Akt/mTOR signaling pathway［J］.J Army Med Univ，2025，47（24）：3009-3021.DOI：10.16016/j.2097-0927.202511049.
唐银，张捷，毛毳，等.高盐饮食通过PI3K/AKT/mTOR通路激活促进代谢功能障碍相关脂肪性肝炎纤维化进展［J］.陆军军医大学学报，2025，47（24）：3009-3021.DOI：10.16016/j.2097-0927.202511049. DOI：

TANG Y，ZHANG J，MAO C，et al.High-salt diet promotes the progression of fibrosis in metabolic dysfunction-associated steatohepatitis via activating the PI3K/Akt/mTOR signaling pathway［J］.J Army Med Univ，2025，47（24）：3009-3021.DOI：10.16016/j.2097-0927.202511049. DOI：

摘要

目的

探究小鼠高脂饮食模型建立的代谢功能障碍相关脂肪性肝炎（metabolic dysfunction-associated steatohepatitis， MASH）在联合高盐饮食背景下对MASH相关肝纤维化进展的影响及潜在的分子机制。

方法

采用C57BL/6J小鼠建立正常饮食小鼠模型、单纯高盐饮食小鼠模型、MASH小鼠模型和MASH联合高盐饮食模型，到达实验终点后取材分别对小鼠肝组织进行苏木素-伊红染色（hematoxylin-eosin staining，H-E）、天狼星红染色（Sirius staining）、免疫组织化学染色（immunohistochemistry，IHC）、油红O染色（Oil Red O）来对小鼠肝脏进行组织学层面评估其脂肪变性程度、脂质沉积及纤维化程度，另外统计分析小鼠肝功和血脂，比较小鼠肝重、体重及其比值变化，对小鼠肝脏组织纤维化标志物及其炎症标志物通过RT-qPCR和Western blot方法进行评估，将肝组织样品送转录组测序后进行生物信息学分析和通过Western blot方法对潜在信号通路进行验证以确认其可能的机制。

结果

通过油红O染色及H

E染色观察可以发现成功建立起代谢功能障碍相关脂肪性肝炎小鼠模型及其联合高盐饮食小鼠模型以及单纯高盐饮食小鼠模型，通过检测血清谷草转氨酶（aspartate transaminase，AST）、谷丙转氨酶（alanine aminotransferase，ALT）、总胆固醇（total cholesterol ，Tch）、甘油三酯（triglyceride，TG）水平可以发现单纯高盐饮食组小鼠在组织学层面与普通饮食组对比无显著差异（

0.05），而MASH联合高盐饮食组较MASH饮食组AST、ALT水平升高更为显著（

0.01），表明MASH背景下的高盐饮食可以加重肝损伤；另外通过天狼星红染色、α-SMA免疫组织化学染色发现MASH联合高盐饮食可以加剧MASH相关的肝纤维化（

0.01），同样MASH联合高盐饮食组在反应纤维化激活相关基因Col1a1、α-SMA、Timp1、Pdgfrβ mRNA水平上表达升高更为显著（

0.01），MASH联合高盐饮食组α-SMA和Fibronectin蛋白表达水平同样上调更为显著，除此之外反应炎症激活相关基因TNF-α、IL-6、IL-1β、TGF-β1在MASH联合高盐饮食组中表达水平明显上调（

0.05），表明MASH联合高盐饮食促进了肝炎及肝纤维化发展；转录组测序分析显示PI3K/AKT/mTOR信号通路激活和TGF-β信号上调更为显著，热图显示纤维化和炎症相关基因显著上调；对富集信号通路相关分子进行Western blot实验验证显示MASH联合高盐饮食组的PI3K/AKT/mTOR信号通路显著激活（P

0.01）。

结论

高盐饮食通过PI3K/AKT/mTOR通路激活促进代谢功能障碍相关脂肪性肝炎纤维化进展。

Abstract

Objective

To explore the impact of high-salt diet (HSD) on the progression of high-fat diet (HFD)-induced metabolic dysfunction-associated steatohepatitis (MASH)-related liver fibrosis

as well as the underlying molecular mechanisms.

Methods

C57BL/6J mice were used to establish the following models: normal diet (Chow)

HSD diet (Chow+8%salt)

MASH diet (MASH)

and MASH diet combined with HSD (MASH+8%salt). At the experience endpoint

the liver tissues were collected and subjected to HE staining

Sirius Red staining

immunohistochemistry (IHC) assay

and Oil Red O staining for histological assessment of steatosis

lipid deposition

and fibrosis. The levels of liver function indicators and serum lipids were measured and statistically analyzed

and the differences in liver weight

body weight

and their ratios were compared across the groups. RT-qPCR and Western blotting were applied to detect the mRNA and protein levels of hepatic fibrosis and inflammatory markers. Liver tissue samples were subjected to transcriptome sequencing

followed by bioinformatics analysis. Potential signaling pathways were validated through Western blotting to confirm the underlying mechanisms.

Results

Oil Red O and HE staining confirmed the successful establishment of the mouse models of MASH

MASH+8%salt

and Chow+8%salt. Serum levels of aspartate transaminase (AST)

alanine aminotransferase (ALT)

total cholesterol (Tch)

and triglyceride (TG) showed no significant differences between the Chow+8%salt and Chow groups (

0.05). In contrast

the MASH+8%salt group exhibited significantly higher AST and ALT levels than the MASH group (P

0.01)

indicating that HSD exacerbates liver injury in the context of MASH. Sirius Red and α-SMA IHC staining revealed that HSD aggravated MASH-related liver fibrosis (

0.01). Similarly

the mRNA levels of fibrosis activation-related genes (Col1a1

α-SMA

Timp1

Pdgfrβ) were more significantly up-regulated in the MASH+8%salt group (

0.01). Protein expression levels of α-SMA and Fibronectin were also markedly elevated. Furthermore

the mRNA levels of inflammation-related genes (TNF-α

IL-6

IL-1β

TGF-β1) were obviously up-regulated in the MASH+8%salt group (

0.05)

suggesting that HSD promotes hepatitis and liver fibrosis progression in MASH. Transcriptome sequencing indicated significant activation of the PI3K/AKT/mTOR signaling pathway and up-regulation of the TGF-β pathway. Heatmap analysis demonstrated marked up-regulation of fibrosis- and inflammation-related genes. Western blotting confirmed significant activation of the PI3K/AKT/mTOR signaling pathway in the MASH+8%salt group (P

0.01).

Conclusion

High-salt diet promotes the progression of hepatitis and liver fibrosis in metabolic dysfunction associated steatohepatitis by activating the PI3K/AKT/mTOR signaling pathway.

关键词

Keywords

references

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High-salt diet promotes the progression of fibrosis in metabolic dysfunction-associated steatohepatitis via activating the PI3K/Akt/mTOR signaling pathway

DOI：10.16016/j.2097-0927.202511049

摘要

Abstract

关键词

Keywords

references