Farhangi Ali, Norouzian Dariush, Mehrabi Mohammad Reza, Chiani Mohsen, Saffari Zahra, Farahnak Maryam, Akbarzadeh Azim
Pilot Biotechnology Department, Pasteur Institute of Iran, No. 69, 12 Farvardin Street, Jomhoori Avenue, 13169-43551 Tehran, Iran.
Indian J Clin Biochem. 2014 Oct;29(4):406-17. doi: 10.1007/s12291-013-0331-2. Epub 2013 May 15.
The objective of this study is to induce experimental diabetes mellitus by streptozotocin in normal adult Wistar rats via comparison of changes in body weight, consumption of food, volume of water, urine and levels of glucose, insulin and C-peptide in serum, between normal and diabetic rats. Intra-venous injection of 60 mg/kg dose of streptozotocin in 250-300 g (75-90 days) adult Wistar rats makes pancreas swell and causes degeneration in Langerhans islet β-cells and induces experimental diabetes mellitus in 2-4 days. For a microscopic study of degeneration of Langerhans islet β-cells of diabetic rats, biopsy from pancreas tissue of diabetic and normal rats, staining and comparison between them, were done. In this process, after collagenase digestion of pancreas, islets were isolated, dissociated and identified by dithizone method and then with enzymatic procedure by DNase and trypsin, the islet cells changed into single cells and β-cells were identified by immune fluorescence method and then assayed by flow-cytometer. Donor tissue in each step of work was prepared from 38 adult male Wistar rats weighted 250-300 g (75-90 days). Transplantation was performed in rats after 2-4 weeks of diabetes induction. In this study, the levels of insulin, C-peptide and glucose in diabetic rats reached to normal range as compared to un-diabetic rats in 20 days after transplantation of islet cells. Transplantation was performed under the cortex of testis as immunoisolated place for islet cells transplantation.
本研究的目的是通过链脲佐菌素诱导正常成年Wistar大鼠患实验性糖尿病，比较正常大鼠和糖尿病大鼠的体重、食物消耗量、饮水量、尿量以及血清中葡萄糖、胰岛素和C肽水平的变化。对体重250 - 300克（75 - 90日龄）的成年Wistar大鼠静脉注射60毫克/千克剂量的链脲佐菌素，可使胰腺肿胀，导致胰岛β细胞变性，并在2 - 4天内诱发实验性糖尿病。为了对糖尿病大鼠胰岛β细胞的变性进行微观研究，对糖尿病大鼠和正常大鼠的胰腺组织进行活检、染色并比较。在此过程中，胰腺经胶原酶消化后分离出胰岛，用双硫腙法进行解离和鉴定，然后通过DNA酶和胰蛋白酶的酶促程序，使胰岛细胞变成单细胞，用免疫荧光法鉴定β细胞，再用流式细胞仪进行检测。工作各步骤中的供体组织取自38只体重250 - 300克（75 - 90日龄）的成年雄性Wistar大鼠。在诱导糖尿病2 - 4周后对大鼠进行移植。在本研究中，胰岛细胞移植后20天，糖尿病大鼠的胰岛素、C肽和葡萄糖水平与未患糖尿病的大鼠相比恢复到正常范围。移植在睾丸皮质下进行，作为胰岛细胞移植的免疫隔离部位。
