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目的观察循环单核细胞亚群在小鼠脑缺血再灌注模型中的动态变化,探讨其与梗死面积及神经功能缺损程度间的关系。方法 30只C57BL/6健康雄性小鼠随机分为大脑中动脉堵塞/再灌注组即缺血再灌注组(MCAO/R组)和假手术组(Sham组)。采用线栓法制备小鼠MCAO/R模型。分别于术前及术后6 h、12 h、第1天、第2天、第3天行神经功能缺损评分,并于术前及术后第1天、第2天、第3天取血进行流式细胞术检测,循环单核细胞分为淋巴细胞抗原6C高表达(Ly6Chigh)及低表达(Ly6Clow)两个亚群,检测各时间点Ly6Chigh及Ly6Clow细胞比例变化。各时间点取小鼠大脑分别进行氯化三苯基四氮唑(TTC)染色、常规HE染色,并经相关性分析探讨单核细胞亚群比例变化与梗死面积及神经功能缺损程度间的关系。结果与基线相比,MCAO/R组小鼠Ly6Chigh单核细胞比例于术后第1天即明显升高,第2天达到最高峰,然后逐渐下降;与Sham组比较,MCAO/R组小鼠各时间点Ly6Chigh单核细胞比例均明显升高。TTC染色结果显示,第2天后梗死范围明显增大。术后各时间点Ly6Chigh单核细胞比例与脑组织梗死面积百分比呈显著正相关,Ly6Chigh单核细胞比例与神经功能缺损评分亦表现显著正相关。结论 MCAO/R模型术后单核细胞亚群动态变化发现,Ly6Chigh亚群于第2天达到高峰,与脑梗死面积变化相关。
Objective To observe the dynamic changes of circulating monocyte subsets in mouse model of cerebral ischemia-reperfusion and to explore its relationship with the infarct size and the degree of neurological deficits. Methods Thirty male C57BL / 6 male mice were randomly divided into MCAO / R group and sham operation group (sham group). Mice MCAO / R model was prepared by thread method. Neurological deficit scores were taken before operation and at 6 h, 12 h, 1 d, 2 d and 3 d after operation respectively. Blood samples were taken before operation and on the 1st, 2nd and 3rd day The flow cytometry was used to detect the changes of Ly6Chigh and Ly6Clow cells at different time points. The circulating monocytes were divided into two subgroups: Ly6Chigh and Ly6Clow. At each time point, the brain of mice was taken for TTC staining and routine HE staining respectively. Correlation analysis was performed to investigate the relationship between the proportion of monocyte subpopulation, infarct size and neurological deficit . Results Compared with the baseline, the proportion of Ly6Chigh monocytes in MCAO / R mice increased significantly on the first postoperative day and reached the peak on the second day, then decreased gradually. Compared with Sham group, the mice in MCAO / R group Ly6Chigh mononuclear cells were significantly increased at all time points. TTC staining results showed that the infarct size increased significantly after the second day. There was a significant positive correlation between Ly6Chigh mononuclear cell percentage and the percentage of infarct area at each time point. Ly6Chigh mononuclear cell percentage was also positively correlated with neurological deficit score. Conclusions The dynamic changes of monocyte subsets after MCAO / R model found that Ly6Chigh subgroup reached its peak on the second day, which was related to the change of infarction area.