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Gene & Protein in Disease Hydrogen sulfide ameliorates NAFLD

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Figure 6. Autophagy protein and lipid droplet protein expression in mouse hepatocytes. (A) Effects of animal fat modeling agent and NaHS treatment on
the AMP-activated protein kinase and mechanistic target of rapamycin pathway in mouse hepatocytes. (B) Analysis of p-mTOR expression; (C) Analysis
of p-AMPK expression; (D) Effects of starvation (Starved), fat modeling agent (EL), and NaHS (EL + NsHS) on LC3 expression in mouse hepatocytes;
(E) LC3 protein expression in mouse hepatocytes; (F) Results of protein expression assays in mouse liver tissue from the control, HFD, and HFD + NaHS
groups; (G) Differential analysis of p-mTOR expression; (H) Differential analysis of p-AMPK expression; (I) Differential analysis of p62 expression;
(J) ADRP protein expression; (K) LC3II/LC3Ⅰ protein expression in mouse liver tissue. Notes: Experimental data are expressed as mean ± SD (n = 3). *P <
0.05, **P < 0.01, and ***P < 0.001, compared with the control group; #P < 0.05, ##P < 0.01, and ###P < 0.001, compared with the EL group.
Abbreviations: NaHS: Sodium hydrosulfide; EL: Edible lard; ADRP: Adipose differentiation-related protein; HFD: High-fat diet.
liver fat content was assessed using triglyceride and total dependent system (Figure 5B). In NAFLD patients, there
cholesterol assay kits. Following the intraperitoneal was a reduction in the liver transcripts of AMPK (PRKAA)
injection of NaHS in HFD mice, a considerable reduction and its downstream targets TSC1/2, whereas transcripts
in hepatic triglyceride and total cholesterol levels was of RHEB, which is upstream of mTOR, were increased.
observed in the NaHS-treated group (Figures 4D and E), In addition, transcripts of autophagy-related genes
indicating the beneficial impact of NaHS on NAFLD in mice. SQSTM1 (p62) and MAP1LC3B2 (LC3) were elevated,
These experimental findings provide substantial evidence whereas transcripts for genes involved in the fusion of
of NaHS’s effectiveness in reducing fat accumulation in the autophagosomes and lysosomes, namely CTSB and CTSL,
liver tissues of NAFLD mice. were decreased (Figure 5C). These findings suggest a
strong association between NAFLD and the transcription
3.5. Transcriptional data analysis reveals NAFLD’s of genes involved in the AMPK-mTOR pathway, the PI3K-
association with the AMPK-mTOR pathway and Akt pathway, and autophagy.
autophagy
3.6. NaHS treatment activates the AMPK-mTOR
Liver transcription data of NAFLD patients (including pathway and autophagy in cells and mice
both steatosis and NASH) and healthy individuals (both
normal weight and obese) were acquired from the GEO Activation of the AMPK-mTOR pathway is characterized
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database. After standardizing the data, the transcripts by increased phosphorylation of AMPK protein and
of differentially expressed genes in liver tissues were decreased phosphorylation of mTOR protein. 50,51
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analyzed. The control groups consisted of individuals According to Tanida et al., LC3 serves as a marker for
with normal weight and those with obesity (Figure 5A). autophagy, with an increased conversion from LC3I to
The transcriptomic data revealed significant associations LC3II indicating enhanced autophagy intensity.
between NAFLD and several pathways, including the In our study, treatment with various concentrations of
PI3K-Akt pathway, sulfur metabolism pathway, and sulfur- NaHS led to a significant increase in AMPK phosphorylation

Volume 3 Issue 3 (2024) 8 doi: 10.36922/gpd.3409