Fig 2: GM cooperates with DEX to antagonize the effect of AMPK inhibitor CC on cell apoptosis, oxidative stress and the inflammatory response in HFD-induced type 2 diabetes mellitus rats. Protein expression of AMPKa1 was (A) determined using western blotting and (B) quantified. Expression of downstream anabolic genes, including SREBP-1c, FAS and DGAT-2 was (C) determined using western blotting and (D) quantified. Expression of catabolic genes of AMPKa1, including CPT-1, PPAR-a and ACA was (E) determined using western blotting and (F) quantified. (G) TUNEL staining was used to evaluate cell apoptosis where normal cells were stained blue and apoptotic cells were stained brown (magnification, ×400). (H) Quantification of apoptotic cells. Concentration of (I) SOD and (J) TNF-a, IL-1ß and IL-6 in serum samples. *P<0.05 vs. ctrl group; #P<0.05 vs. high-fat group; &P<0.05 vs. CC group; ^P<0.05 vs. CC + DEX group. GM, germacrone; DEX, dexmedetomidine; AMPK, AMP-activated protein kinase; CC, compound C; AMPKa1, AMP-activated protein kinase a1; SREBP-1c, sterol regulatory element binding protein-1c; FAS, fatty acid synthase; DGAT-2, diacylglycerol acyltransferase-2; CPT-1, carnitine palmitoyltransferase-1; PPAR-a, peroxisome proliferator-activated receptor-a; ACA, acyl coenzyme A; TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling; SOD, superoxide dismutase; TNF-a, tumor necrosis factor-a; IL, interleukin; Ctrl, control.

Fig 3: GM improves the effect of DEX in regulating AMPKa1, downstream lipid metabolism indicators and anabolic genes in HFD-induced type 2 diabetes mellitus rats. Protein expression levels of AMPKa1 were (A) determined by western blotting and (B) quantified. Expression of downstream anabolic genes, including SREBP-1c, FAS and DGAT-2 was (C) determined by western blotting and (D) quantified. Expression of catabolic genes of AMPKa1, including CPT-1, PPAR-a and ACA was (E) analyzed in liver tissues by western blot analysis and (F) quantified. Relative protein expression was normalized to ß-actin. *P<0.05 vs. ctrl group; #P<0.05 vs. HFD group; &P<0.05 vs. DEX group. GM, germacrone; DEX, dexmedetomidine; AMPKa1, AMP-activated protein kinase a1; SREBP-1c, sterol regulatory element binding protein-1c; FAS, fatty acid synthase; DGAT-2, diacylglycerol acyltransferase-2; CPT-1, carnitine palmitoyltransferase-1; PPAR-a, peroxisome proliferator-activated receptor-a; ACA, acyl coenzyme A; ctrl, control.

Fig 4: Effects of HBP on the expression of genes related with cholesterol and triglyceride metabolism in the liver: (A) Relative mRNA level of LDLR and DGAT2; (B) relative protein levels of LDLR and DGAT2; (C) representative bands of immunoblot. Values not sharing a letter in common are significantly different, p < 0.05.

Fig 5: Liver gene and protein expression at 22 weeks. (A) mRNA expression by RT-qPCR of main transcription factors (Chrebp-1, Srebp-1, Ppara, Ppar?), (B) fatty acids synthesis (Hmgcr, Soat1, Acc1, Fasn, Scd1, Dgat2), (C) uptake/efflux fatty acids (Cd36, Cd204, Fatp2, Abca1, Abcg1), (D) oxidative stress (Nrf2, Hmox-1, Sod1, Catalase, Pgc-1a, Drp-1), (E) Inflammation (Ccl2, Ccl5, Cxcl10, Cx3cl1, Il-15, Tlr4), and (F) fibrosis (Tgfß, Ctgf, Grem1) was evaluated. Fold changes of the target gene was normalized by of their respective housekeeping gene 18s ribosomal subunit, (G) Protein expression of lipogenic enzymes by western blot was evaluated. Fold changes of proteins levels in BTBR ob/ob vs. BTBR WT (n-fold = 1) normalized by ß-Actin and images of their respective Western Blot. Data are shown as scatter dot plots and mean ± SEM of each group (n = 6 mice/group); * p < 0.05, ** p < 0.01 vs. BTBR WT.