Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Feb 10;14(3):1265-1279.
doi: 10.18632/aging.203817. Epub 2022 Feb 10.

Hesperidin suppresses ERS-induced inflammation in the pathogenesis of non-alcoholic fatty liver disease

Qi Xie  1   2 , Shuqing Gao  1 , Min Lei  3 , Zengning Li  2   4
Affiliations

Hesperidin suppresses ERS-induced inflammation in the pathogenesis of non-alcoholic fatty liver disease

Qi Xie et al. Aging (Albany NY). .

Abstract

Objective: The current study aimed to establish a non-alcoholic fatty liver disease (NAFLD) model using HFD-fed SD rats and FFA-stimulated human THP-1 cells to examine whether hesperidin (HSP) plays a role in endoplasmic reticulum stress (ERS)-induced inflammation in the pathogenesis of NAFLD.

Methods: Oil red O staining was used to determine the effect of HSP on hepatic steatosis in rat liver tissues. Differentially expressed genes (DEGs) were subjected to functional enrichment analysis by bioinformatics. Western blotting was used to detect the protein expression of GRP94, ATF6, ATF4, p-PERK, p-IRE1α, IL-1β, IL-6, and TNF-α in liver tissues and THP-1 cell lines, and the expression of GRP94 and p-PERK in vitro was detected through immunofluorescence staining.

Results: HSP significantly decreased the weight gain, hepatic steatosis but not serum lipid profile and suppressed the serum levels of inflammatory factors in HFD-fed rats. It was revealed by bioinformatics analysis that the inflammatory response and IRE1α activation were enriched signaling pathways in NAFLD. The expression of ERS-related biomarkers, GRP94, ATF6, ATF4, p-PERK and p- IRE1α, was significantly suppressed by HSP in vivo and in vitro. Moreover, the inflammatory markers, including IL-1β, IL-6, and TNF-α, were also decreased by HSP in vivo and in vitro. Immunofluorescence staining exposed that the expression of GRP94 and p-PERK was decreased by HSP in vitro.

Conclusion: HSP may suppress ERS-induced inflammation in the pathogenesis of NAFLD.

Keywords: endoplasmic reticulum stress; hesperidin; inflammation; non-alcoholic fatty liver disease.

PubMed Disclaimer

Conflict of interest statement

CONFLICTS OF INTEREST: The authors declare no conflicts of interest related to this study.

Figures

Figure 1
Figure 1
HSP suppressed HFD-induced body weight gain and hepatic steatosis but not serum lipid profile. SD rats were fed with CD or HFD for 4 months and were randomly divided into CD + NS group, CD + HSP group, HFD + NS group, and HFD + HSP group. (A) The body weight of the rat was evaluated. Data were presented as mean ± SEM. **P < 0.05. (B) The levels of serum TC, TG, and LDL, FFA were examined (C) the levels of serum IL-1β, IL-6, TNF-α. Data were presented as mean ± SEM. (D) Histological analysis of hepatic steatosis stained with Oil Red O staining.
Figure 2
Figure 2
Identification of DEGs in two NAFLD-related GEO datasets. NAFLD-related gene expression profiles GSE48452 and GSE89632 were employed to detect the DEGs between NAFLD patients and healthy donors. (A) The results of pre-standardization and post-standardization based on GSE48452 were exhibited. (B) The results of pre-standardization and post-standardization based on GSE89632 were exhibited. (C) Heap map exhibited the upregulated and downregulated DEGs in GSE48452. X-axis exhibited samples. Y-axis exhibited the genes. The left dendrogram exhibited gene clustering. The upper dendrogram exhibited the sample clustering. (D) Heap map exhibited the upregulated and downregulated DEGs in GSE89632. X-axis exhibited samples. Y-axis exhibited the genes. The left dendrogram exhibited gene clustering. The upper dendrogram exhibited the sample clustering.
Figure 3
Figure 3
Functional annotation and GSEA analysis of DEGs. (A, B) GO and KEGG enrichment analysis of DEGs associated with NAFLD-related GSE48452. (C, D) GO and KEGG enrichment analysis of DEGs associated with NAFLD-related GSE89632. (E, F) GSEA of DEGs associated with NAFLD-related GSE48452 and GSE89632.
Figure 4
Figure 4
HSP could attenuate ERS-induced inflammation in the liver. (A) Immunofluorescence staining was performed to examine the expression of GRP94 in liver tissues. (B) Western blotting analysis was performed to examine the expression of GRP94, ATF6, ATF4, p-PERK, and p-IRE1α in liver tissues. Data were presented as mean ± SEM. **P < 0.05, HFD + HSP vs. HFD; &&P < 0.05, ND + HSP vs. ND. (C) Western blotting analysis was performed to examine the expression of IL-1β, IL-6, and TNF-α. Data were presented as mean ± SEM. **P < 0.05, HFD + HSP vs. HFD; &&P < 0.05, ND + HSP vs. ND.
Figure 5
Figure 5
HSP attenuates ERS in human THP-1 cells. As described in the method, FFA-stimulated human THP-1 cells were employed to establish the NAFLD model. Thapsigargin is a well-described ERS inducer successfully used in various cell lines. (A) Western blotting analysis was performed to examine the expression of GRP94, ATF6, ATF4, p-PERK, and p-IRE1α in human THP-1 cells. (B) Western blotting analysis was performed to examine the expression of IL-1β, IL-6, and TNF-α.
Figure 6
Figure 6
HSP attenuates ERS in human THP-1 cells. As described in the method, FFA-stimulated human THP-1 cells were employed to establish the NAFLD model. Thapsigargin was used to simulate the in vitro ERS model. (A) Immunofluorescence staining was performed to examine the expression of GRP94 and p-PERK with or without HSP. (B) Relative fluorescence intensity of GRP94 and p-PERK. Data were presented as mean ± SEM. **P < 0.05, FFA + HSP vs. FFA.
Figure 7
Figure 7
HSP suppressed macrophages modulated the lipid metabolism related proteins expression in hepatocytes cells. FFA-stimulated THP-1 cells treated with HSP or Thapsigargin, and the associated cell supernatant were used to stimulate hepatocytes. The lipid metabolism associated proteins: SREBP-1/-2 and C/EBP β were tested by Western blotting, as shown in (A) and (B). Data were presented as mean ± SEM. **P < 0.05 control vs. HSP.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Musso G, Gambino R, Cassader M. Recent insights into hepatic lipid metabolism in non-alcoholic fatty liver disease (NAFLD). Prog Lipid Res. 2009; 48:1–26. 10.1016/j.plipres.2008.08.001 - DOI - PubMed
    1. Gaggini M, Morelli M, Buzzigoli E, DeFronzo RA, Bugianesi E, Gastaldelli A. Non-alcoholic fatty liver disease (NAFLD) and its connection with insulin resistance, dyslipidemia, atherosclerosis and coronary heart disease. Nutrients. 2013; 5:1544–60. 10.3390/nu5051544 - DOI - PMC - PubMed
    1. Yang CQ, Shu L, Wang S, Wang JJ, Zhou Y, Xuan YJ, Wang SF. Dietary Patterns Modulate the Risk of Non-Alcoholic Fatty Liver Disease in Chinese Adults. Nutrients. 2015; 7:4778–91. 10.3390/nu7064778 - DOI - PMC - PubMed
    1. Feng RN, Du SS, Wang C, Li YC, Liu LY, Guo FC, Sun CH. Lean-non-alcoholic fatty liver disease increases risk for metabolic disorders in a normal weight Chinese population. World J Gastroenterol. 2014; 20:17932–40. 10.3748/wjg.v20.i47.17932 - DOI - PMC - PubMed
    1. Nobili V, Day C. Childhood NAFLD: a ticking time-bomb? Gut. 2009; 58:1442. 10.1136/gut.2009.184465 - DOI - PubMed

Publication types

MeSH terms