| Background and Purpose:It has been believed that it is a critical management to promote intracerebral hematoma to absorb faster, to control the brain edema and ameliorate the injured tissues around the hematoma after intracerebral hemorrhage (ICH) up to now. However, there is few method available to treat the hematoma itself. It is even short of efficacious medicine in promoting the hematoma to liquidize and absorb, except for extracting the hematoma by surgery. Therefore, we referred to the previous therapy studies of ICH in experimental and clinical aspects with hirudo (leech) and its polypharmacy. Our aims of this study is to explore the native absorption mechanisms of experimental intracerebral hematoma in Wistar rats, mainly to investigate the effect of hirudo extract liquor (HEL) on intracerebral hematoma in rats and its potential pharmaceutical mechanisms from the body, tissues, cells to molecular biology level by the method of combined with in vivo and in vitro, also investigate related complications after HEL therapy, trying to provide more experimental bases for its clinical use in future.Methods: We established the experimental ICH model in rats by stereotaxical injecting quantitative collagenase(0.7U collagenaseⅦ) into their left caudate nuclei. In vitro, brain microvascular endothelial cells (BMEC) and astrocytes (As) from Wistar rat cerebral cortex were cultured respectively. In the present study, hematoma volume, infarction volume in perihematoma tissues (PHT), neurological severity scores, brain water content (BWC), biochemical index, including tissue-typeplasminogen activator (tPA) and plasminogen activator inhibitor (PAI) activity, and tPA, PAI-1and D-dimer (D-D) contents in plasma, PHT and cultured supernatants, as well as variation of semi-quantification of tPA, PAI-1 mRNA levels by RT-PCR in PHT and in cultured rat BMEC, were determined respectively. Fbg, PT, aPTT levels in plasma were simultaneously determined too. Histopathological changes including light microscope, electronic microscope and angiography were also observed. At the same time, the effects of HEL on them were delineated. BWC was calculated by drying-weighing method. Neurological deficits were examined with modified Bederson's method, infarction area with TTC staining. Activities of tPA, PAI were measured by colorimetric assay. Contents of tPA, PAI-1 and D-D were respectively determined by using specific ELISA. Local capillaries were observed by angiography with ink-dextran infusion. Expression of HSP70,TGFβ-1 and tPA protein in PHT,BMEC and As was investigated by immunohistochemistry.Cell activity or injury was assayed by measuring mitrochondrial function (MTT assay). Cell morphology under the inverted phase contrast microscope was observed and cell activity with MTT assay was measured after exposing BMEC to a selected concentration of HEL ranging from 0.0625 to 8 mg/μl. The survival of As was investigated in vitro by exposing the cells to a selected concentration of thrombin ranging from 0.1 to 100U/ml and of HEL ranging from 0.25 to 4 mg/μl by observing cell morphology and measuring the lactate dehydrogenase (LDH) from damaged cells, a marker of cell death. Results:(1) There was ischemic damage in PHT during the acute phase of experimental ICH. (2) With the time past, intracerebral hematoma became smaller and smaller, but it was remarkably small till the 10th day, leaving incomplete absorption in some rats till the 14th day. The tPA amount, activity and its mRNA expression in PHT were increased, aPTT lengthened on the 3rd day and the 6th day after experimental ICH, showing dose-dependent in the experiment. However, PAI-1 amount, PAI relative activity and PAI-1 mRNA expression were not obviously depressed. (3) HEL could quicken intracerebral hematoma in rats to liquidize and absorb, significantly decreasing the hematoma volume and ischemic area on the 6th and 10th day, as well asreduce brain edema, markedly decreasing BWC both at 48 hours and on the 4th day. In addition, the recovery of neurological...
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