Glucose-lowering effect of DLBS3233 is mediated through phosphorylation of tyrosine and upregulation of PPARγ and GLUT4 expression

Figures & data

Table 1 Primers used to amplify region of interest in PCR

Gene name	Primer sequences	PCR product (bp)
PI3 kinase	Sense: 5′-GAA GAC TGA GAG GCC ACG AC-3′	180
	Antisense: 5′-CTT GTA ATC CAT GGC GTC CT-3′
Akt	Sense: 5′-ACT GTT CAG AGA GGC CAG GA-3′	181
	Antisense: 5′-CGG TTG CCT ACT GGT TCA AT-3′
PPARγ	Sense: 5′-GCT GTG CAG GAG ATC ACA GA-3′	229
	Antisense: 5′-CCC CTC CAT AAA CTC ACC AA-3′
GLUT4	Sense: 5′-ACT CTT GCC ACA CAG GCT CT-3′	174
	Antisense: 5′-AAT GGA GAC TGA TGC GCT CT-3′
Adiponectin	Sense: 5′-GTT GCA AGC TCT CCT GTT CC-3′	192
	Antisense: 5′-TCT CCA GGA GTG CCA TCT CT-3′
Resistin	Sense: 5′ GCT GAG GGT CTG GAA ATG AA-3′	223
	Antisense: 5′ GGC CAG CCT GGA CTA TAT GA-3′
Beta actin (internal control)	Sense: 5′ AGA GGG AAA TCG TGC GTG AC 3′	138
	Antisense 5′ CAA TAG TGA TGA CCT GGC CGT 3′

Abbreviations: PCR, polymerase chain reaction; PPARγ, peroxisome proliferator-activated receptor gamma; PI3, phosphatidylinositol-3 .

Figure 1 DLBS3233 increases glucose uptake in 3T3 Swiss albino cells compared with untreated cells. A) Glucose uptake level of 3T3 Swiss albino preadipocytes and adipocytes in normal and insulin-resistant condition. B) Effect of DLBS3233 5 μg/mL and pioglitazone 0.02 μM on glucose uptake in normal and insulin-resistant 3T3 Swiss albino adipocytes. C) DLBS3233 5 μg/mL enhances glucose uptake activity of 3T3 Swiss albino insulin-resistant adipocytes as potently as pioglitazone in time-dependent manner. Results are mean value ± standard deviation of two independent experiments.

Figure 2 Peroxisome proliferator-activated receptor gamma gene expression after administration of Cinnamomum burmanii extract (A), Lagestroemia speciosa extract (B), and DLBS3233 (C) at various concentrations.

Figure 3 Effect of DLBS3233 5 μg/mL on expression of tyrosine phosphorylation (A). The effect of DLBS3233 5 μg/mL and pioglitazone 0.02 μM in enhancing phosphatidylinositol-3 kinase (B) and Akt (C) gene expression in 3T3 Swiss albino preadipocytes. The effect of DLBS3233 1–5 μg/mL in enhancing glucose transporter 4 gene expression in 3T3 Swiss albino preadipocytes (D). Effect of DLBS3233 and pioglitazone with or without insulin in enhancing glucose transporter 4 gene expression (E), and also total glucose transporter 4 protein level (F).

Notes: Results are mean value ± standard deviation of two independent experiments. *P < 0.05 indicates significant difference compared with controls.
Abbreviation: AU, arbitrary unit.

Figure 4 DLBS3233 increases transcription level of adiponectin (A), and decreases transcription level of resistin (B) in 3T3 Swiss albino preadipocytes.

Notes: Results are mean value ± standard deviation of two independent experiments. *P < 0.05 indicates a significant difference compared with control.

Figure 5 Effect of DLBS3233 9 mg/kg body weight on the levels of glucose (A), postprandial glucose (B), fasting glucose (C), insulin (D), triglycerides (E), total cholesterol (F), low-density lipoprotein (G), and high-density lipoprotein (H) in insulin-resistant Wistar rats, and comparison of fasting glucose and insulin level (I) and further expressed as homeostasis model assessment (J).

Notes: Results are mean value ± standard deviation of two independent experiments. *P < 0.05 indicates a significant difference compared with GF group.
Abbreviation: GF, glucose and fructose.