Male albino Wistar rats weighing 180-220 g were used throughout the study. The rats were housed in an air-
conditioned room and had free access to water and a pellet diet (Pashu Aahar Kendra, Varanasi, India). All the
experimental procedures were carried out in accordance with internationally accepted guidelines for the care and
use of laboratory animals. All animal procedures used were in strict accordance with the Committee for the Purpose
of Supervision on Animal Experiments under the Animal Welfare Departments and Ministry of Environment and
Forest, Govt. of India, India. All experimental protocols were approved by the U niversity of A llahabad A nimal
Experimentation Ethics Committee. Diabetes was induced by a single intraperitonial injection of freshly prepared STZ at a
dose of 55 mg/kg body weight in 0.1 M citrate buffer (pH 4.5) to overnight fasted rats. After 3 days of STZ administration,
rats with marked hyperglycemia in terms of fasting blood glucose (FBG) and postprandial glucose (PPG) [FBG > 250
mg/dL and PPG > 350 mg/dL were used in the study. Our previous study had demonstrated 200 mg/kg dose of
the aqueous extract as the most effective dose[14]. Therefore, this dose was used in the present study to evaluate its
antioxidant potential in vivo in diabetic rats. Rats were divided into 5 groups of six rats each. Group 栺consisting of
normal rats, served as normal control, treated with vehicle(distilled water) only whereas, Group 栻 comprised normal
animals, treated with 200 mg/kg of the extract. Group 栿 had diabetic animals, serving as diabetic control, received
vehicle only. Diabetic animals of Group 桇 were treated with a known synthetic drug, Glipizide at a dose of 2.5 mg/kg.
D iabetic animals of G roup 桋 received leaves aqueous extract at a dose of 200 mg/kg daily up to 21 days. After 21
days treatment, the rats were deprived of food overnight and sacrificed by cervical dislocation. B lood samples
were collected from the heart. Key organs including heart, brain, liver, kidney, pancreas and spleen were quickly
removed, washed immediately with ice-cold saline, dried and were used or stored at -80 曟 for further experiments.
10 % ( w/v ) homogenates of the organs were prepared in phosphate buffer (100 mM, pH 7.4) containing 150 mM KCl.
The homogenates were centrifuged at 9 000 伊 g for 30 min. Clear supernatants were used in the estimation of all the
biochemical parameters.