| ObjectivesPrevious studies have suggested that inflammatory reactions have neurotoxic effects in the pathophysiological process of cerebral hemorrhage,Recent studies have found that inflammation may play a bidirectional role in the pathophysiological process of intracerebral hemorrhage,And the bidirectional effect may be related to different types of inflammatory cells or different subtypes of the same cell.Current research in animal experiments found that the composition of immune cells in peripheral blood and hematoma fluid of acute cerebral hemorrhage has changed significantly,and the proportion or number of some cell components is related to the prognosis of cerebral hemorrhage,there is still no systematic clinical study on the components of cellular immune cells in peripheral blood and hematoma fluid in the acute stage of cerebral hemorrhage.To evaluate the changes of cellular immune components in peripheral blood and hematoma fluid of patients with acute cerebral hemorrhage and their relationship with clinical prognosis.This study examined the changes of cellular immune components in peripheral blood and hematoma fluid in acute stage of cerebral hemorrhage,and evaluated the relationship between them and 90-day prognosis of patients with cerebral hemorrhage.It is hoped that through this study,therapeutic targets that may be related to the prognosis of cerebral hemorrhage will be screened out..Method35 patients with acute ICH underwent minimally invasive evacuation of hematoma in the neurosurgery of the Fifth Affiliated Hospital of Zhengzhou University and the people’s Hospital of Gongyi city were enrolled as the study group from January 1,2016,to October 31,2017.Fifty-five age-matched healthy controls were also recruited.Peripheral blood samples(10 m L)were collected once from ICH patients before the surgery.And drainage fluid samples(7-10 m L)from the hematoma.Peripheral blood samples(10 m L)were drawn from control subjects on the day after they signed the consent.Flow cytometry was used to detect the percentage of lymphocyte,monocyte,granulocyte,T helper cells(Th),cytotoxic T lymphocytes(CTL)and B lymphocyte,natural killer cells(NK),Th17 cells,and regulatory T cells(Treg),T cell receptor(TCR)T cells,T cell receptor(TCR)T cells,dendritic cells(DC),M1 macrophages,M2 macrophages,M1 microglia,and M2 microglia.National Institutes of Health Stroke Scale(NIHSS),Glasgow Coma Scale(GCS),time from symptom onset to MIS,lesion volume and other clinical data were recorded.Neurological scores(m RS)were recorded at 90 days after onset.The changes of immune cells in peripheral blood and hematoma fluid of patients with acute cerebral hemorrhage were analyzed,and the relationship between the proportion of immune cells and the degree of brain injury at admission and the prognosis at 90 days was evaluated.Results1.Changes of immune cell count and proportion in peripheral blood and hematoma fluid of patients with acute cerebral hemorrhage.Compared with the control group,the absolute counts of white blood cells,monocytes,granulocytes and the percentage of granulocytes,Treg cells,Th17 cells were all significantly increased in peripheral blood.whereas the absolute count of lymphocytes and the percentage of lymphocytes,M1 and M2 macrophages were significantly decreased in peripheral blood of ICH patients.Compared with tthe peripheral blood of ICH patients,the percentage of lymphocytes,cytotoxic T cells,dendritic cells,M1 and M2 macrophages were significantly higher,while the percentage of granulocytes,Th cells and B lymphocytes were significantly lower in the hematoma.2.Comparison of immune cells in peripheral blood and hematoma of patients with acute cerebral hemorrhage in different groupsUnivariate analysis showed that the percentage of TCRαβ T cells was lower(P=0.028),and the percentage of M1 macrophages higher(P=0.038),in the hematoma of female patients than in that of male patients.Compared with patients younger than 60 years old,patients younger than 60 years old had lower proportion of Treg cells in peripheral blood and higher proportion of Th17 cells,M1 macrophages and M1 microglia in hematoma fluid.The percentage of Treg cells in peripheral blood of patients with GCS < 7 was each lower than that of patients with GCS > 7.The percentage of Th cells in the peripheral blood was significantly lower at 12-30 hours than it was within the first 12 hours after ICH.the percentage of CTLs was higher in the hematoma of patients with lesion volume > 50 m L than in that of patients with lesion volume ≤ 50 m L.3..Analysis of the correlation between the degree of brain injury and the proportion of immune cells in peripheral blood and hematoma blood in patients with acute cerebral hemorrhage at admissionThe lesion volume was positively correlated with the percentage of CTLs but negatively correlated with the percentage of M2 microglial cells in hematoma fluid.GCS score was positively correlated with the time from symptom onset to MIS(r=0.458,p =0.006)negatively correlated with NIHSS scorer(-0.352,p = 0.038)on admission,After adjustment for the time from symptom onset to MIS and NHISS score,the GCS score and positively correlated with the percentage of T cells in the hematoma fluid.NIHSS score was positively correlated with age,and negatively correlated with GCS score on admission.After adjustment for age and GCS score,the NIHSS score marginally correlated with the percentage of M1-like macrophages(r=0.34,P=0.056)in the peripheral blood of ICH patients.4.The correlation between the proportion of immune cells in peripheral blood or hematoma fluid and the prognosis of 90 days after acute ICH.Univariate analysis revealed that patients with a favorable outcome(m RS≤3)on day 90 were significantly younger than those with an unfavorable outcome(m RS>3)(56.4 years vs.67.7 years;P=0.014).The NIHSS score of patients with a favorable outcome was significantly lower than that of patients with an unfavorable outcome(19 vs.24;P=0.013).Univariate analysis of immunocytes and inflammatory cytokines indicated that,when compared to patients with an unfavorable outcome,patients with a favorable outcome had significantly higher percentages of monocytes and Treg cells,and significantly lower percentages of granulocytes in peripheral blood.In addition,the percentages of M1-like macrophages,and M1-like microglia were significantly lower in the hematoma of patients with a favorable outcome than in that of patients with an unfavorable outcome(all P<0.05).Bivariate logistic regression analysis showed that the increase of M1 macrophage proportion in hematoma fluid(OR = 5.23,P = 0.044)after excluding the effects of age,NHISS score and systolic blood pressure was related to the adverse prognosis of patients with cerebral hemorrhage at 90 days.Conclusion1.Intracerebral hemorrhage has a significant impact on the peripheral immune system.The immune cell components in the hematoma fluid may originate from the ruptured blood vessel extravasation or migrate to the peripheral immune cells around the hematoma through the blood-brain barrier.2.Immunocyte components in peripheral blood or hematoma fluid may have predictive value for the prognosis of cerebral hemorrhage,such as the increase of M1 macrophage proportion in hematoma fluid is associated with adverse prognosis. |
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