يك روش ساده براي اندازه گيري هيستامين و فعاليت آنزيم هيستيدين دكربوكسيلاز در مغز موش آزمايشگاهي

The content of the histamine in most mammalian tissues is very low. Thus , presentation a simple and sensitive method to assay the histamine content in crude extracts tissues is required . In this work ,we presented a simple method for the measurement of the histamine. 2 mg of rat brains was homogenized in 5 ml of 0.5 M hCIO4 .The homogenates were centrifuged (3000*g , 3 min )and supernatants were adjusted to ph 5 with 0.4 M HCI. Then the precipitate was removed by centrifugation (3000*g, 5 min) , and the supernatants were put into a micro centrifuge filter tube . ( exclusion molecular weight 10000,Millipor ) and centrifuged at 3000 *g, for 5 min and supernatants was put into a tube .The histamine was measured spectrophotometrically using 2,4,6-trinitrobenzen sulfonate ,at 420 nm. One unit of enzyme activity is defined as the amount of enzyme which will produce 1 micro mole of the histamine per min at 25° C. The results presented in this work indicated that the highest content of histamine level and histidine decarboxylase were in hypothalamus. The results obtained here clearly showed that this method is simple, sensitive , reliable and could be applied to assay histamine content and histidine decarboxylase activity in the brain.