| The hypoxic ischemic brain injury is a complex pathophysiological reaction process. There are still some obscure issues in the mechanism. In the process of acute cerebral infarction, some inflammatory factors, like IL-10, IL-6, TNF-α, TGF-β1, CRP and so on, are expressed at high levels in the brain tissue suffered with acute hypoxic ischemic injury, which indicate that the formed complexity of inflammatory factors stimulated by the hypoxic ischemic injury might play a role in the repair process. Some current studies have shown that the exposure of some antigens from the damaged brain tissue triggers immunological and inflammatory reaction in which cell immunity occupies a leader place. During the process, inflammatory factors induce the expression of adhesive molecules and chemotactic factors which cause the migration, infusion and aggregation of inflammatory cells into the surrounding place beside the damaged brain tissue. In the development of inflammation, the disorder of inflammatory reaction leads to the second hit to the brain. However, some evidences of the middle cerebral artery blocked rat models tests have demonstrated T cells are in a state of immunological depression in which Treg cells are at a reactive high level after cerebral infarction. Thus, the immunological depression might be attributed to the repairment of the damaged brain. But in the clinical practice, we observed that the rate of pulmonary and urinary infections is at a high level for patients suffered from massive cerebral infarction rather than patients with lacunar or normal cerebral infarction. According to the above observation, we designed this research to clarify some distinctions of immunological and inflammatory reaction in patients with different types of cerebral infarctions and the relationship between the immunological and inflammatory reaction.To test and analyze the proportion of Treg cells and levels of associated inflammatory factors in the peripheral blood of patients in the early stage of acute cerebral infarction.1. Different groups designationWe selected 45 cases diagnosed as the acute cerebral infarction by the DWI sequence of MRI in the different groups of cerebral infarction. The volume of cerebral infarction was determined by the equation "length X width X thickness/2" of lesion. According to the volume of lesions, cases were designed as three groups, the mall infarction group (<5 cm3), the normal infarction group (5-10 cm3) and the massive infarction group (>10 cm3).15 TIA (transient ischemic attack) cases without positive lesions in the DWI sequence of MRI were designed as the Control group.2. Laboratory testsThe fasting peripheral blood was sampled from patients at different time points, 24h,72h and 6 days after the onset of cerebral infarction. Serums and mononuclear cells were separated from blood by using the method of density gradient separation. The levels of hs-CRP, IL-10, TGF-β1 were determined by ELISIA tests, and the proportions of Treg, IL-10+ and TGF-β1 cells in the lymphocytes were determined by detecting CD4+/CD25+/Foxp3+, IL10+ and TGF-β1+ by using flow cytometry.3. Statistic analysisData were analyzed by t-tests and linear correlation analysis.1. The analysis of serum hs-CRPAfter the acute cerebral infarction 24 hours、72 hours、6 days,the levels of hs-CRP in the serum were determined, (P<0.01). There is no significant statistic difference in comparison between TIA and small cerebral infarction groups (P>0.05). The level of hs-CRP was downgraded after 72 hours.There are significant statistic differences in the comparison among the small infarction group, normal infarction group and massive infarction group.2. The results of flow cytometric tests(1) The proportion of CD4+ T cells in the lymphocytes of different groups:in the comparison between the TIA and small cerebral infarction groups, there is no significant difference (P=0.52). Compared with the normal group, data of the small cerebral infarction group make no significant differences(P=0.3). There is a significant statistic difference in the comparison between the normal cerebral infarction and massive cerebral infarction groups (P<0.05)(2) The proportion of CD4+CD25+Foxp3+ T cells in the lymphocytes of different groups:in the comparison between the TIA and small cerebral infarction groups, there is no significant difference (P=0.85). Compared with the normal group, data of the small cerebral infarction group make a significant differences (P<0.01). There is a significant statistic difference in the comparison between the normal cerebral infarction and massive cerebral infarction groups (P<0.001)(3) The proportion of TGF-β1+ T cells in the lymphocytes of different groups:in the comparison between the TIA and small cerebral infarction groups, there is no significant difference (P=0.9). Compared with the normal group, data of the small cerebral infarction group make no significant differences(P=0.3). There is a significant statistic difference in the comparison between the normal cerebral infarction and massive cerebral infarction groups (P<0.05)(4) The proportion of IL-10+ T cells in the lymphocytes of different groups:in the comparison between the TIA and small cerebral infarction groups, there is no significant difference (P=0.56). Compared with the normal group, data of the small cerebral infarction group make a significant differences (P<0.05). There is a significant statistic difference in the comparison between the normal cerebral infarction and massive cerebral infarction groups (P<0.05)3. The results of ELISIA test(1) After the acute cerebral infarction 24 hours, the levels of TGF-1, in the comparison between the TIA and small cerebral infarction groups, there is no significant difference (P=0.08). Compared with the normal group, data of the small cerebral infarction group make no significant differences (P=0.08). There is a significant statistic difference in the comparison between the normal cerebral infarction and massive cerebral infarction groups (P<0.001)(2) After the acute cerebral infarction 24 hours, the levels of IL-10 in the serum, significant difference (P=0.8). Compared with the normal group, data of the small cerebral infarction group make no significant differences(P=0.7). There is a significant statistic difference in the comparison between the normal cerebral infarction and massive cerebral infarction groups (P<0.001)4. The analysis of the correlation between the serum hs-CRP and proportions of Treg cells, TGF-β1+ cells, and IL-10+ cellsThere is significant positive linear correlation between the proportion of Treg+ cells in the 24 hours time point and the serum hs-CRP in the 72 hours time point (r=0.87, P <0.05). The similar results were also observed in the comparison of TGF-β1 vs hs-CRP (r=0.78, P<0.05) and IL-10 vs hs-CRP (r=0.51, P<0.05).1. Compared with the TIA control group, the level of serum hs-CRP was enhanced after the acute cerebral infarction 24 hours, and at the peak in the time point of 72 hours; as time goes, the level of hs-CRP was downgraded after 72 hours; the change of serum hs-CRP in the small cerebral infarction group is not significant.2. Compared with the TIA control group, after the onset of cerebral infarction 24 hours, Treg cells proliferated and released TGF-β1 and IL-10 at a high level in the massive cerebral infarction group; the capacity of Treg cells releasing IL-10 rather than TGF-β1 was enhanced in the normal infarction group; for the small cerebral infarction group, there is no significant change in the capacity of Treg cells releasing TGF-β1 and IL-10.3. Compared with the TIA control group, after the onset of cerebral infarction 24 hours, the levels of serum TGF-β1 and IL-10 were dramatically enhanced in the massive infarction group, rather than the small and normal infarction group.4. There is significant positive linear correlation between the proportions of Treg+ cells, TGF-β1+ and IL-10+ cells in the 24 hours time point and the serum hs-CRP in the 72 hours time point. |
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