Fig 1.
Sketch map which showed the process of the animal experiment.
Table 1.
Primary antibodies used for Western Blotting (WB) and immunohistochemistry (IHC).
Table 2.
The primer sequence for genes.
Table 3.
Art ameliorated diabetic symptoms and fasting blood glucose, HbA1c, and serum insulin levels in T1DM mice.
Fig 2.
Artemether ameliorated liver injury in T1DM mice.
(A) Serum ALT level. (B) Serum AST level. (C) HE staining of mouse liver sections, the scale bars are located in the lower left corner. The scale bars from left to right are 50 µm, 20 µm, and 10 µm. (D-F) Western blotting for GSTP1, and SOD2(acetyl K68) in the liver. ALDH2 serves as an internal reference. The results were indicated as the means ± SEMs (n = 6–8), ***p < 0.001 vs. Control group; ###p < 0.001 vs. the T1D group.
Fig 3.
The effects of artemether on the structure of liver mitochondria.
(A) The structure of mitochondria in the liver of mice under transmission electron microscopy, showing nuclei (N), endoplasmic reticulum (ER) and mitochondria(M), the red dashed boxes represent the incomplete magnified view, while the red solid boxes denote the complete magnified view. The scale bars are located in the lower left corner. (B-C) Western blotting for COXⅣ in the liver. ALDH2 serves as an internal reference. The results were indicated as the means ± SEMs (n = 6-8). *p < 0.05 vs. Control group; ##p < 0.01 vs. the T1D group.
Table 4.
The content of mitochondrial metabolites in the liver. The results were indicated as the means ± SEMs. (n = 6–8), *p < 0.05 and **p < 0.01 vs. Control group; #p < 0.05 vs. the T1D group.
Fig 4.
Artemether regulated mitochondrial function in the liver.
(A) The ratio of ATP to ADP in the liver. (B) The ratio of NAD+ to NADH in the liver. (E) mRNA expression levels of Idh3b in the liver. (F) mRNA expression levels of Nd1 in the liver. (C, G, H) Western blotting for ND2 and MTCO2 in the liver. (K) IHC for MTCO2 in liver, the scale bars are located in the lower left corner. (D, I, J) Western blotting for IDH3A and IDH3B in the liver. ALDH2 serves as an internal reference. The results are indicated as the means ± SEMs (n = 4-8), *p < 0.05, **p < 0.01, and ***p < 0.001 vs. Control group; #p < 0.05, ##p < 0.01 and ###p < 0.001 vs. the T1D group.
Fig 5.
Artemether prevented hepatic fat mobilization and fatty acid activation, and regulated triglycerides in T1DM mice to normal levels.
(A) Serum TG levels in the mice. (B) Hepatic TG levels in the mice. (C) mRNA expression levels of Atgl, Fatp2 and Acsl1 in the liver. (D-F) Western blotting for ATGL and HSL in the liver. (G-J) Western blotting for FATP2, ACSS3 and ACSM2A in the liver. ALDH2 serves as an internal reference. The results were indicated as the means ± SEMs(n = 4-8), *p < 0.05, * *p < 0.01 and ***p < 0.001 vs. Control group; #p < 0.05, ##p < 0.01 and ###p < 0.001 vs. the T1D group.
Fig 6.
Artemether inhibited liver fatty acidβ oxidation in T1DM mice.
(A) Hepatic stearoyl-CoA content in the mice. (B) Hepatic stearoyl-L-carnitine content in the mice. (C) Hepatic FAD levels in the mice. (D-I) Western blotting for CPT1A, CPT2, CACT, CROT, and ACADM in the liver. ALDH2 serves as an internal reference. (J) mRNA expression levels of Cpt1a, Cact, Crot, Acads, and Acadl in the liver. The results were indicated as the means ± SEMs (n = 4–8), *p < 0.05, **p < 0.01, and ***p < 0.001 vs. Control group; #p < 0.05, ##p < 0.01, and ###p < 0.001 vs. the T1D group.
Fig 7.
Artemether increased de novo synthesis of fatty acids in the liver of T1DM mice.
(A) and (D-F) Western blotting for ACLY, ACC, and FASN in the liver. ALDH2 serves as an internal reference. (B) IHC for FASN in the liver, the scale bars are located in the lower left corner. (C) mRNA expression levels of Fasn in the liver. The results were indicated as the means ± SEMs (n = 4–8), *p < 0.05 and **p < 0.01 vs. Control group; #p < 0.05, ##p < 0.01, and ###p < 0.001 vs. the T1D group.
Fig 8.
Artemether promoted the oxidative utilization of glucose in the liver of T1DM mice.
(A) Hepatic pyruvate content in T1DM mice. (B) Hepatic lactic acid content in T1DM mice. (C) IHC for PKLR in the liver, the scale bars are located in the lower left corner. (D) mRNA expression levels of Gck in the liver. (E) mRNA expression levels of Pdk1 in the liver. (F-I) Western blotting for GCK, PKLR and LDHA in the liver. (J-M) Western blotting for PDK1, p-PDH(Ser293), and PDH in the liver. ALDH2 serves as an internal reference. The results were indicated as the means ± SEMs (n = 4–8) *p < 0.05, **p < 0.01 and ***p < 0.001 vs. Control group; #p < 0.05, ##p < 0.01, and ###p < 0.001 vs. the T1D group.
Fig 9.
Artemether inhibited glycogenolysis and gluconeogenesis in T1DM mice.
(A) mRNA expression levels of Pcx in the liver. (B) mRNA expression levels of Pck1 in the liver. (C, F, G) Western blotting for p-PYGL(Ser15) and PYGL in the liver. (D, H, I) Western blotting for PCK1 and PCK2 in the liver. (E) IHC for p-PYGL(Ser15) in the liver, the scale bars are located in the lower left corner. ALDH2 serves as an internal reference. The results were indicated as the means ± SEMs (n = 4–8), *p < 0.05, **p < 0.01, and ***p < 0.001 vs. Control group; #p < 0.05 and ###p < 0.01 vs. the T1D group.
Fig 10.
The schematic diagram illustrates hepatic glucose metabolism, lipid metabolism, and mitochondrial metabolism in the context of Art intervention in T1DM mice.
In T1D, glycolysis is reduced, while gluconeogenesis, glycogenolysis, fatty acid β-oxidation/activation are enhanced, and de novo fatty acid synthesis is inhibited—leading to mitochondrial dysfunction and liver injury. In contrast, T1D+Art intervention increases glycolysis, decreases gluconeogenesis/glycogenolysis, inhibits fatty acid β-oxidation/activation, and promotes de novo fatty acid synthesis, thereby alleviating mitochondrial dysfunction and reducing liver injury. Art modulates key enzymes/metabolites in glucose and lipid metabolism. For glucose metabolism, Art affects glycogenolysis (via p-PYGL), gluconeogenesis (via PCK1/2, PCX), and glycolysis (via GCK, PKLR, LDHA). For lipid metabolism, it influences triglyceride (TG) hydrolysis (via ATGL, HSL), fatty acid activation (via ACSS3, ACSM2A), de novo fatty acid synthesis (via ACLY, ACC, FASN), and TCA cycle (via IDH3A/B). Red/blue indicators respectively show Art-upregulated/downregulated molecules in T1D mice, highlighting Art’s role in restoring metabolic homeostasis and mitigating T1D-related liver injury.