| Project/Area Number |
58870111
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Biological pharmacy
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| Research Institution | Tohoku University |
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Principal Investigator |
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| Project Period (FY) |
1983 – 1985
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| Project Status |
Completed (Fiscal Year 1985)
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| Budget Amount *help |
¥5,300,000 (Direct Cost: ¥5,300,000)
Fiscal Year 1985: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1984: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1983: ¥3,500,000 (Direct Cost: ¥3,500,000)
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| Keywords | Atniinflammatory Agent / Prostaglandin / Leukotriene / Fluorescein-label / Bovine serum albumin / 空気嚢法 |
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| Research Abstract |
1. Aim of the project: It is to develop a rational method for evaluating antiinflammatory efficacy of a drug in that the evaluation could be done based on the assay of pathophysiological aspects of inflammatory reaction together with the assay of chemical mediators in the site of inflammation. The principle of the assay may be well materialized by using experimental models of the inflammation of air pouch type which has been developed in this laboratory.
2. Methods and the results:
(1) Almost complete extraction of fluorescein-labeled bovine serum albumin from rat dermis was attempted with successful results by digesting the tissue with NaOH followed by clearing it with the aid of ethanol. The supernatant can be assayed in a fluorospectrometer to determine plasma exudation in dermal inflammatory response.
(2) An HPLC method was investigated to determine leukotrienes in inflammatory pouch fluids. Sep-pak C-18 cartridge was proved to be effective in the extraction of leukotrienes from the pouch fluids. The leukotriene faction from the cartridge was loaded on a reverse phase column first. Each leukotriene fraction collected was again loaded on a straight phase column for the final determination. This was successful to separate and measure leukotriene B4, while recovery rates of peptidoleukotrienes were unsatisfactory. The assay method has been thus established for the measurement of leukotriene B4 for individual rats with the air pouch inflammation.
(3) Our trial for HPLC measurement of PGE was unsuccessful, because radioimmunoassay has been shown to be more convenient to measure PGE in the pouch fluid.
(4) The present proposal for the rational evaluation of antiinflammatory agents is to mesure plasma exudation in combination with the PGE level in the exudate and to measure leukocyte infiltration in combination with the chemotactic acitivity of the exudate.
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