Characterization of extracted ellagic acid
Ellagic acid was derived from strawberry. The characterization has been demonstrated by UV. UV–Vis absorbance spectra values against incident wavelength (190–1100) nm for all investigated samples. It can observed a sharp characteristic absorption peak in case extract EA at (275 nm ≈ 4.47 eV) which corresponds to the band gap energy and attributed to π→π* transitions.
Table 1 displayed that; significant elevation (p < 0.05) in HBA1C, ALT, AST, and ALP levels in diabetic rats compared to control group. But, co- or/and post treated diabetic rats with EA revealed a significant depletion (p < 0.05) in HBA1C, ALT, AST, and ALP levels compared to diabetic group.
Table 1
Changes of HBA1C and liver enzymes in different groups.
Parameters | Control | Ellagic acid | Diabetic | Co-treated | Post-treated |
HBA1C (mmol/mol) | 5.4 ± 0.12 | 5.0 ± 0.09 | 7.0***±0.29 | 5.6**±0.25 | 5.17****±0.08 |
ALT U/L) | 41.0 ± 1.5 | 40.7 ± 1.8 | 80.8****±1.11 | 60.0****±1.6 | 41.0****±1.5 |
AST (U/L) | 120.7 ± 3.4 | 120.3 ± 1.7 | 169.8****±4.09 | 132.7****±4.26 | 124.0****±3.05 |
ALP (U/L) | 180.4 ± 10.8 | 181.8 ± 11.4 | 536.0****±9.7 | 473.8****±10.7 | 276.5****±9.3 |
The significance of difference was analyzed by one – way ANOVA (compare all vs. Diabetic group) by computer program. Data was expressed as means ± SE. one – way ANOVA was significant at P < 0.05.
Effect of ellagic acid on levels of lipid profile of diabetic rats.
Table (2) displayed that; elevation (p < 0.05) in total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholestrol (HDL-C) and very low density lipoprotein-cholesterol (VLDL-C) levels while serum low density lipoprotein-cholestrol (LDL-C) levels was declined in diabetic animals compared to the control. In treated groups (G4&G5), EA reduced HBA1C, ALT, AST, ALP, TC, TG, LDL-C and VLDL-C levels whereas serum HDL-C concentrations were enhanced.
Table 2
Effects of ellagic acid extract on lipid profile of diabetic rats. Mean ± SE are used to express the data. Each group contain 10 rats.
Parameters | Control | Ellagic acid | Diabetic | Co-treated | Post-treated |
Cholesterol (mg/dl) | 172.4 ± 0.08 | 169.3 ± 1.11 | 189.8****±1.16 | 179***±1.7 | 174****±2.31 |
Triglyceride (mg/dl) | 174 ± 2.5 | 172.8 ± 1.9 | 223.0****±1.8 | 195.3****±2.02 | 175****±1.6 |
HDL-C (mg/dl) | 34.9 ± 1.9 | 33.25 ± 1.7 | 55.5****±1.3 | 57.7#±0.88 | 59.0#±1.5 |
LDL-C (mg/dl) | 100.5 ± 1.28 | 99.53 ± 1.07 | 94.9**±0.84 | 87.27***±1.16 | 76.63****±0.98 |
The significance of difference was analyzed using one – way ANOVA (compare all vs. Diabetic group) by computer program. Data was expressed as means ± SE. one – way ANOVA was significant at P < 0.05.
Liver Histopathology
Hepatic sections in male rats in the control and EA groups displayed hepatocytes with normal shape and structure as hepatocytes are polygonal in shape with round nuclei and eosinophilic cytoplasm (Fig. 2A and B). But, Liver sections in diabetic group displayed hepatotoxicity detected by marked inflammatory cells, hepatic cords degeneration, many of apoptotic cells, as well as congested blood sinusoids (Fig. 2C and D). Liver sections in co-treated diabetic rats with EA groups showed mild to moderate inflammatory cells and congested blood sinusoids (Fig. 2E). However, hepatic sections in post-treated diabetic rats with EA groups a good improvement in hepatocytes with mild degeneration, and mild inflammatory cells (Fig. 2F).
Impact Of Ellagic Acid On Proliferating Cell Nuclear Antigen (Pcna) Expression
Figure 3 revealed that the changes in anti-PCNA immunoreactivity in hepatic sections in different groups were strong positive reaction for anti-PCNA immunoreactivity in hepatic sections were observed in control and EA groups (Fig. 3A,B), mild to moderate in diabetic group (Fig. 3C,D), moderate in co-treated and strong in post-treated groups (Fig. 3E,F).
Impact Of Ellagic Acid On Interleukin-6 (Il-6) Expression
Figure 4 revealed the changes in anti-IL-6 immunoreactivity in hepatic sections in different groups. Mild positive reactions for anti-IL-6 immunoreactivity in hepatic sections were observed in control and EA groups (Fig. 4A,B). While; strong reactions were detected in diabetic group (Fig. 4C,D), Nevertheless; a moderate reactions in co-treated and moderate to mild reactions in post-treated groups were detected (Fig. 4E,F).
Real Time-pcr Analysis Of Hepatic Insulin Like Growth Factor-1 And Klotho Genes
The RT-PCR data as showed in Fig. 5A,B indicated that, in diabetic group the expression of IGF-1 genes were significantly (P < 0.0001) downregulated compare to the control rats, but the expression of klotho genes were non significantly (P < 0.05) downregulated compare to the control rats. Over expression of IGF-1 and klotho genes were induced when diabetic rats treated with ellagic acid (G4,G5) compare to diabetic untreated rats. The results of correlation analysis showed that the expression levels of IGF1and klotho were strongly positive correlated in the liver of rats with diabetes (r = 0.939, P < 0.01), as showed in Fig. 6.