All chemicals, solvents and standards (purity > 98%) used in the study were commercially obtained from (Sigma-Aldrich, Germany) and were of analytical grade without further purification. The positive standards, including glibenclamide (purity ≥ 99%) and tramadol hydrochloride (purity ≥ 99%) were also obtained from Sigma-Aldrich, Germany. The purity of the tested compound 3 was ca. 95%, HPLC.
Swiss-albino male healthy mice (25.0–30.0 g) of Wistar-strain were obtained from the University animal house (BAU, Lebanon). The mice were housed for one week before experimentation under standard controlled conditions of temperature (25.0 ± 1.0 °C) with twelve hrs dark-light cycle, and free access to water and commercially obtained mice-feed…show more content… Serum catalase (CAT) levels (kU/l) were measured subchronically using the modified method elucidated before in the literature  (Table 1b). Moreover, lipid peroxidation was measured by thiobarbituric acid test that was modified from a previously described method . Reduced glutathione (GSH) was also assessed by a modified method illustrated before .
Assessment of diabetic neuropathy in mice The diabetic mice provoked peripheral neuropathy after 8 weeks of induction of T1DM with alloxan , which was manifested from a significant hyperalgesia and tactile-allodynia accompanied with decrease in pain and withdrawal threshold responses in tail flick, hot plate and Von Frey filaments. Only diabetic mice with persistent hyperglycemia (HbA1c >8%) showed clear thermal hyperalgesia (significant sensory loss of heat sensitivity below 10S) after 8 weeks of alloxan administration [1,10-12]. Tail withdrawal, hot plate and Von Frey filament were performed to monitor the extent of DN within control, positive control, utilizing 10 mg/kg tramadol (TRA), and test groups of mice. Thermal hyperalgesia was assessed by hot plate and tail withdrawal assays. The mechanical responses were evaluated by Von Frey filaments. These tests were repeated for 8 weeks post-administration in control, positive control and test groups of