Immunologic process of diabetes induced by sub-dose of streptozotocin.

Research Project

Project/Area Number	60571047
Research Category	Grant-in-Aid for General Scientific Research (C)
Allocation Type	Single-year Grants
Research Field	Biological pharmacy
Research Institution	University of Tokushima
Principal Investigator	KAWADA Jun University of Tokushima, Professor, 薬学部, 教授 (10035537)
Co-Investigator(Kenkyū-buntansha)	NISHIDA Mikio University of Tokushima, AssociateProfessor, 薬学部, 助教授 (10035561) KOBAYASHI Shigeru University of Tokushima, Professor, 薬学部, 教授 (70035533)
Project Period (FY)	1985 – 1986
Project Status	Completed (Fiscal Year 1986)
Budget Amount *help	¥1,500,000 (Direct Cost: ¥1,500,000) Fiscal Year 1986: ¥300,000 (Direct Cost: ¥300,000) Fiscal Year 1985: ¥1,200,000 (Direct Cost: ¥1,200,000)
Keywords	Monolayer culture of pancreatic B-cells / Streptozotocin / 3-O-methyl-streptozotocin / Experimental diabetes / Cellular immunity / エチリデングルコーズ
Research Abstract	1. A system of pancreatic B-cell culture was established. The pancreatic tissues from new-born rats were digested with 0.2% trypsin and 0.1% collagenase, and the dispersed cells were cultured in medium 199 including 16.7 mM glucose. The cultured cells maintained good morphological appearances and a high rate of insulin secretion under these conditions for over 10 days. 2. A new diabetogenic streptozotocin analogue, 3-O-methyl-streptozotocin, was synthesized. The critical point of this synthesis was nitrosylation of an immediadte precursor compound of the final product. This difficulty was overcomed by using 40% acetic acid solution of NaN <O_2> . These conditions enabled us to establish a stable synthetic procedure of this new biologically active compound. 3. In vivo effects of 3-O-methyl-streptozotocin and streptozotocin were compared in rats. A variety of parameters tested suggested that the cytotoxicity for pancreatic B-cells of 3-O-methyl-streptozotocin was milder than that of streptozotocin. 4. A consecutive administration of sub-dose of streptozotocin in rats induced delayed hyperglycemia and low glucose tolerance. We examined wether an immunologic mechanism was involved in this phenomenon. However, such possibility was excluded by using ethylidene glucose as a probe. Therefore, species difference was thought to be an important factor for the onset of immunologic diabetes in experimental animals.