| Co-Investigator(Kenkyū-buntansha) |
MASUDA Midori KANNSAIMEDICAL UNEVERSITY,FACULTY OF MEDICINE,ASSISTANT, 医学部, 講師 (50173753)
KOMIYAMA Yutaka KANNSAIMEDICAL UNEVERSITY,FACULTY OF MEDICINE,ASSISTANT, 医学部, 講師 (40140264)
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| Research Abstract |
We have been studying the role of central nervous system in the production of endogenous digitalislike factor (s) (EDLF) by using experimental animals for more than 15 years. Recently, we found that the adrenal medulla also contained digitalislike immunoreactivity in rats. Therefore, we thought that the cell-line of adrenomedullary origin could produce (EDLF). In fact, we found that PC-12 cells which is originated from pheochrocytoma cells produced ouabainlike immunoreactive substance, which was characterized by repeated chromatography. Furthermore, production of EDLF in this culture system was obviously but slightly augmented by a growth factor (since this evidence has not been published, we do not want to indicate the material here). In the present study period, we collected a great amount of culture medium after stimulating with the growth factor. After passing serveral kinds of elution columns, the EDLF was purified, and the fraction which contains ouabainlike immunoreactivity was analyzed by liquid chromatography with mass spectrometry. As the result, the substance was identified to be ouabain. This must be the first evidence that mammals can endogenously produce digitalis, because the other reports always have had the possibility that the identified material is of the diet origin. However, some other materials which showed the different chromatographic characteristics were there. The analysis is now going on. Furthermore, we identified the ouabain-binding protein in plasma, which was a fragment of IgG.
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