2.2. Animals, induction of diabetes and study protocol
Wistar rats (Rattus norvegicus) of either sex, weighing between 200 and 225 g
were procured from the animal house, Delhi Institute of Pharmaceutical Sciences
and Research, (Protocol No: DIPSAR/IAEC/11/2008). All experimental procedures
and animal care followed the institutional guidelines and Association for Research
in Vision and Ophthalmology statement for the use of animals in eye research. All
animals were maintained under adequate conditions at an ambient temperature of
21 ± 2 C, and were subjected to12 h light and dark cycle. They were fed with standard
rat chow and water ad libitum.
Animals were randomly divided into normal control and streptozotocin-induced
(45 mg/kg i.p) diabetic groups. The diabetic rats were further divided into
diabetic control and TC (250 mg/kg, oral) treated diabetic groups. Dose was selected
on the basis of literature survey. Each group consisted of 15 animals.
Treatment was initiated three days after STZ induction. Animals showing blood
glucose level above 250 mg/dl were selected for the study. Blood sugar was monitored
weekly till the end of the study. Photographs of anterior and posterior chamber
of the eye were taken at adequate intervals to evaluate the lenticular and
fundoscopic changes.
After 24 weeks of diabetes, animals were sacrificed with deep anesthesia, blood
was collected in EDTA-coated vacuutainers and retinae were isolated, blotted,
weighed and stored for further analysis. Kidney, pancreas, heart and liver were also
isolated and fixed for light microscopy. Retinal homogenate was prepared in phosphate
buffer, pH 7.4 for biochemical estimations. All estimations were done in
duplicate.
2.2. Animals, induction of diabetes and study protocolWistar rats (Rattus norvegicus) of either sex, weighing between 200 and 225 gwere procured from the animal house, Delhi Institute of Pharmaceutical Sciencesand Research, (Protocol No: DIPSAR/IAEC/11/2008). All experimental proceduresand animal care followed the institutional guidelines and Association for Researchin Vision and Ophthalmology statement for the use of animals in eye research. Allanimals were maintained under adequate conditions at an ambient temperature of21 ± 2 C, and were subjected to12 h light and dark cycle. They were fed with standardrat chow and water ad libitum.Animals were randomly divided into normal control and streptozotocin-induced(45 mg/kg i.p) diabetic groups. The diabetic rats were further divided intodiabetic control and TC (250 mg/kg, oral) treated diabetic groups. Dose was selectedon the basis of literature survey. Each group consisted of 15 animals.Treatment was initiated three days after STZ induction. Animals showing bloodglucose level above 250 mg/dl were selected for the study. Blood sugar was monitoredweekly till the end of the study. Photographs of anterior and posterior chamberof the eye were taken at adequate intervals to evaluate the lenticular andfundoscopic changes.After 24 weeks of diabetes, animals were sacrificed with deep anesthesia, bloodwas collected in EDTA-coated vacuutainers and retinae were isolated, blotted,ชั่งน้ำหนัก และเก็บไว้สำหรับการวิเคราะห์เพิ่มเติม ไต ตับอ่อน หัวใจ และตับได้แยก และถาวรสำหรับแสงแบบไมโครสโค Homogenate จอประสาทตาถูกเตรียมในฟอสเฟตบัฟเฟอร์ ค่า pH 7.4 ประเมินทางชีวเคมี ทำทั้งหมดประมาณในที่ซ้ำกัน
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