EFFECT OF METHYLPREDNISOLONE ON THE DEVELOPMENT OF HEMORRHAGE AFTER CEREBRAL ISCHEMIA: DEVELOPMENT OF A PROTOCOL FOR HEMORRHAGE ANALYSIS

Abstract

Ischemic strokes, caused by artery occlusion, can be treated using thrombolysis with tissue plasminogen activator (tPA), which restores blood flow and prevents further brain damage. Unfortunately, one of the side effects of tPA is symptomatic hemorrhagic transformation (SHT), which can be fatal. As a result, many drugs that may prevent hemorrhaging in the brain are being studied. Methylprednisolone, a steroid, is being studied at the Veteran's Affairs Medical Center in Augusta, Georgia. The effects of drugs such as methylprednisolone are determined by comparing blood content in the brains of control and treated animals using qualitative methods, because currently there are no quantitative methods being used to measure blood content. A quantitative method is needed because the effects of the drugs may not be detected using qualitative measurements. Two possible methods for quantitatively measuring blood content in the brain are currently being investigated: the Enzyme-Linked Immunosorbent Assay (ELISA) test and the spectrophotometric hemoglobin assay. The hemoglobin assay is preferred for the methylprednisolone study because it requires less time and fewer chemical reagents. The focus of this study was to determine if the hemoglobin assay would be a suitable method for measuring blood in a study investigating the effects of methylprednisolone and similar drugs. The results of the study indicate that the assay is not a satisfactory quantitative technique for measuring hemoglobin in the brain because the hemoglobin cannot be adequately purified and measured. Also, lipids create turbidity in the samples, which can cause artifacts. Therefore, the ELISA test should be looked at as a possible technique for quantitatively determining blood content because it may be more specific and sensitive to hemoglobin.