
Melatonin and/or rowatinex attenuate streptozotocin-induced diabetic renal injury in rats
Tarek K Motawi, Samia A Ahmed, Manal A Hamed, Shohda A El-Maraghy, Wessam M Aziz
Journal of Biomedical Research ›› 2019, Vol. 33 ›› Issue (2) : 113-121.
Melatonin and/or rowatinex attenuate streptozotocin-induced diabetic renal injury in rats
The study aimed to explore the prophylactic effect of melatonin, rowatinex; a naturally occurring renal drug, and its combination on diabetic nephropathy in type 2 diabetic rats. Diabetes was induced by intraperitoneal injection of a single dose of streptozotocin (50 mg/g body weight). Three days before diabetes induction, rats were daily treated with melatonin, rowatinex and their combination continuously for 8 weeks. Evaluation was done through measuring blood urea nitrogen (BUN), serum uric acid, serum creatinine, urine creatinine, creatinine clearance, nitric oxide (NO), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), total antioxidant capacity (TAC), kidney injury molecule-1 (KIM-1), heat shock protein-70 (HSP-70), caspase-3, transforming growth factor β1 (TGFβ1), DNA degradation by the comet assay and total protein contents. Histopathologic study was also done for the kidney and the pancreas. Drastic changes in all measured parameters of the diabetic rats were observed. Treatment with melatonin and rowatinex showed amelioration to variable degrees. In conclusion, melatonin showed the most potent effect on protecting rats from deleterious action of diabetic nephropathy followed by its combination with rowatinex.
diabetic nephropathy / melatonin / rowatinex / histopathology
Tab.1 Protective effect of different drugs on kidney function parameters in rats with diabetic nephropathy |
Groups | BUN (mg/dL) | Serum uric acid (mg/dL) | Serum creatinine (mg/dL) | Urine creatinine (mg/dL) | Creatinine clearance (mL/minute) |
---|---|---|---|---|---|
Control | 26.48f±2.07 | 1.31f±0.07 | 0.42g±0.04 | 42.52ab±8.91 | 1.26a±0.23 |
Control Rowatinex | 35.32b±2.02 | 1.68e±0.13 | 0.53d±0.05 | 42.94ab±7.76 | 0.992c±0.07 |
Control Melatonin | 29.24cde±1.34 | 1.75 de±0.109 | 0.44fg±0.06 | 41.28ab±11.43 | 0.89c±0.11 |
Control (Rowatinex+ Melatonin) | 26.73f±1.48 | 1.64e±0.18 | 0.54d±0.06 | 40.84b±10.24 | 1.11ab±0.13 |
Diabetic | 61.86a±2.02 | 3.89a±0.26 | 2.67 a±0.18 | 13.86d±8.98 | 0.14d±0.04 |
Prophylactic (Rowatinex) | 31.10c±2.22 | 2.04 c±0.104 | 0.60c±0.06 | 34.20c±5.49 | 0.86c±0.09 |
Prophylactic (Melatonin) | 26.68f±0.81 | 2.21b±0.09 | 0.48ef±0.05 | 46.38 a±10.63 | 0.87c±0.10 |
Prophylactic (Rowatinex+ Melatonin) | 36.66b±2.48 | 2.32 b±0.14 | 0.53d±0.05 | 41.6ab±7.84 | 1.17a±0.82 |
• Data are mean±SD of ten rats in each group. • Statistical analysis are done using one way analysis of variance (ANOVA) using Co Stat Computer program accompanied with least significance level (LSD) between groups at P <0.05. • Unshared superscript letters are significant values between groups at P<0.0001. |
Tab.2 Protective effect of different drugs on oxidative stress markers in rats with diabetic nephropathy |
Groups | NO (µmol/g tissue) | MDA (µg/mg protein) | SOD (µg/mg protein) | GSH (µg/mg protein) | TAC |
---|---|---|---|---|---|
Control | 39.18f±3.95 | 70.68g±2.89 | 0.82a±0.07 | 2.21a±0.12 | 2.26a±0.176 |
Control Rowatinex | 41.46ef±4.61 | 70.20g±5.04 | 0.77b±0.08 | 2.10b±0.22 | 2.05b±0.058 |
Control Melatonin | 43.30e±6.8 | 73.20g±5.22 | 0.68c±0.06 | 1.77f±0.06 | 2.02bc±0.09 |
Control (Rowatinex+ Melatonin) | 51.27d±13.09 | 74.54g±6.38 | 0.696c±0.06 | 2.01a±0.13 | 2.052b±0.13 |
Diabetic | 88.92a±6.59 | 109.65a±7.50 | 0.53d±0.06 | 1.52h±0.057 | 1.21g±0.06 |
Prophylactic (Rowatinex) | 57.94c±5.94 | 78.98f±6.76 | 0.67c±0.07 | 1.92de±0.09 | 1.94de±0.06 |
Prophylactic (Melatonin) | 56.06c±4.98 | 84.48e±9.41 | 0.71c±0.08 | 1.84ef±0.11 | 2.046b±0.13 |
Prophylactic (Rowatinex+ Melatonin) | 57.86c±4.12 | 85.90de±6.69 | 0.73bc±0.12 | 2.02bcd±0.22 | 1.79f±0.08 |
• Data are mean±SD of ten rats in each group. • Statistical analysis are done using one way analysis of variance (ANOVA) using Co Stat Computer program accompanied with least significance level (LSD) between groups at P<0.05. • Unshared superscript letters are significant values between groups at P<0.0001. |
Tab.3 Protective effect of different drugs on kidney injury indices in rats with diabetic nephropathy |
Groups | KIM-1 (pg/mL) | HSP-70 (pg/100 mL) | Caspase-3 (pg/mL) | TDF-b (pg/mL) |
---|---|---|---|---|
Control | 83.39h±22.5 | 1.80i±0.35 | 47.31g±11.60 | 115.24h±23.58 |
Control Rowatinex | 97.36h±19.69 | 2.01h±0.23 | 38.60gh±3.99 | 125.16h±23.06 |
Control Melatonin | 97.22h±21.03 | 1.68i±0.21 | 33.32h±3.45 | 88.91I±5.26 |
Control (Rowatinex+ Melatonin) | 101.9h±26.84 | 2.29g±0.24 | 48.84g±3.31 | 114.24h±26.26 |
Diabetic | 1047.5a±96.72 | 6.89a±0.46 | 362.76a±30.48 | 579.82a±5.09 |
Prophylactic (Rowatinex) | 703.39d±26.08 | 4.84d±0.25 | 292.55b±4.41 | 471.00c±12.37 |
Prophylactic (Melatonin) | 778.52c±17.54 | 4.76d±0.31 | 239.79e±17.46 | 426.38e±32.86 |
Prophylactic (Rowatinex+ Melatonin) | 719.45d±10.39 | 4.844d±0.12 | 259.04d±5.05 | 438.72e±14.67 |
• Data are mean±SD of ten rats in each group. • Statistical analysis are done using one way analysis of variance (ANOVA) using Co Stat Computer program accompanied with least significance level (LSD) between groups at P<0.05. • Unshared superscript letters are significant values between groups at P<0.0001. |
Tab.4 Protective effect of different drugs on DNA degradation and protein contents in rats with diabetic nephropathy |
Groups | DNA tail length (µm) | % of tailed DNA | Tail moment (Unit) | Tissue total protein (mg/g) | Serum total protein (g/dL) |
---|---|---|---|---|---|
Control | 1.502i±0.49 | 1.492j±0.33 | 2.35I±1.22 | 8.14abc±0.81 | 6.05cd±0.44 |
Control Rowatinex | 1.89g±0.08 | 2.14h±0.022 | 4.014h±0.18 | 6.76d±0.84 | 6.18cd±1.052 |
Control Melatonin | 1.682h±0.25 | 1.73i±0.22 | 3.016I±0.84 | 7.81bc±0.58 | 5.46d±0.38 |
Control (Rowatinex+ Melatonin) | 2.23f±0.36 | 2.45g±0.30 | 5.31g±1.51 | 8abc±0.58 | 6.83bc±0.28 |
Diabetic | 5.30a±0.21 | 5.14a±0.31 | 27.47a±2.74 | 5.10e±0.48 | 10.37a±3.20 |
Prophylactic (Rowatinex) | 3.816b±0.17 | 3.86cd±0.067 | 14.80c±0.59 | 8.52a±0.64 | 4.09e±1.23 |
Prophylactic (Melatonin) | 3.97b±0.18 | 4.14b±0.036 | 16.33b±0.86 | 8.40ab±0.43 | 6.22cd±0.96 |
Prophylactic (Rowatinex+ Melatonin) | 3.59c±0.13 | 3.73d±0.48 | 12.92d±0.577 | 7.74c±0.70 | 6.05cd±0.44 |
• Data are mean±SD of ten rats in each group. • Statistical analysis are done using one way analysis of variance (ANOVA) using Co Stat Computer program accompanied with least significance level (LSD) between groups at P<0.05. • Unshared superscript letters are significant values between groups at P<0.0001. |
Fig.1 Electrophoretic pattern of DNA fragmentation in the renal tissues of diabetic rats. Control rats received normal saline (A), melatonin (B), rowatinex (C), or melatonin plus rowatinex (D), and diabetic rats received normal saline (E), prophylactic melatonin (F), rowatinex (G), or melatonin plus rowatinex (H). Minimal breakage and movement of DNA are seen in diabetic rats prophylactically treated with melatonin. |
Fig.2 Histopathological sections of rats kidney and pancreas A: Kidney section of control rats shows the renal cortex of the renal corpuscle with normal glomeruli (black arrow), normal pattern of proximal (red arrow) and distal convoluted (yellow arrow) tubules. B: Pancreatic section of control rats shows atrophic pancreatic islets of Langerhan's as shown by arrow. C: Kidney section of control rats treated with rowatinex shows the renal cortex of the renal corpuscle with normal glomeruli (black arrow), normal pattern of proximal (red arrow) and distal convoluted tubules (blue arrow) with congested blood vessels (green arrow). D: Pancreatic section of control rats treated with rowatinex drug shows regular pancreatic islets of Langerhans as shown by arrow. E: Kidney section of control rats treated with melatonin shows the renal cortex of the renal corpuscle with mild cellularity, mild obliterated capsular space (black arrow), mild thick wall of the proximal tubules (red arrow) and distal convoluted tubules (yellow arrow). F: Pancreatic section of control rats treated with melatonin shows normal size pancreatic islets of Langerhan's with regular shape as shown by arrow. G: Kidney section of control rats treated with rowatinex and melatonin shows the renal cortex of the renal corpuscle with normal glomeruli (black arrow), normal pattern of the proximal (red arrow) and distal convoluted tubules (yellow arrow). H: Pancreatic section of control rats treated with rowatinex and melatonin shows hypertrophy of b-cells of islets of Langerhan's as shown by arrow. I: Kidney section of diabetic rats shows the renal cortex with corpuscles of high cellularity and obliterated capsular space (black arrow). The proximal and distal convoluted tubules show disrupted epithelia (red and yellow arrow). J: Pancreatic section of diabetic rats shows atrophic pancreatic islets of Langerhan's as shown by arrow. K: Kidney section of diabetic rats prophylactically treated with rowatinex shows the renal cortex of the renal corpuscle with normal glomeruli (black arrow), normal pattern of the proximal (red arrow) and distal convoluted (yellow arrow) tubules. L: Pancreatic section of diabetic rats prophylactically treated with rowatinex shows atrophic pancreatic islets of Langerhan's, as shown by arrow. M: Kidney section of diabetic rats prophylactically treated with melatonin shows the renal cortex of renal corpuscle with moderate cellularity and mild obliterated capsular space (black arrow), thick wall of the proximal (red arrow) and convoluted (yellow arrow) tubules. N: Pancreatic section of diabetic rats prophylactically treated with melatonin shows almost normal size pancreatic islets of Langerhan's with regular shape as shown by arrow. O: Kidney section of diabetic rats prophylactically treated with rowatinex and melatonin shows the renal cortex of the renal corpuscle with moderate cellularity, mild obliterated capsular space (black arrow) and normal pattern of the proximal (red arrow) and distal convoluted (yellow arrow) tubules. P: Pancreatic section of diabetic rats prophylactically treated with rowatinex and melatonin showed regular pancreatic islets of Langerhan's as shown by arrow (H&E, ×400). A representative photograph is shown from each group (n = 5). |
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Supplementary files
JBR-16028-OF-TM_suppl_1 (467 KB)
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