目的 探讨再殖小胶质细胞对小鼠脑出血(ICH)神经功能恢复的影响及其机制.方法 采用C57BL/6J成年雄性小鼠,脑立体定向注射胶原酶Ⅳ制备ICH模型.分组为假手术组、单纯ICH组和再殖小胶质细胞组.在ICH后第3天和第14天,采用改良神经功能缺损评分(mNSS)和转角试验评估小鼠神经功能缺损,实时荧光定量聚合酶链反应分析血肿周围脑组织白细胞介素(IL)-6和IL-1β等促炎性细胞因子水平.组间比较采用t检验和单因素方差分析.结果 与假手术组比较(0.38±0.48、50.00±8.88),ICH 后第 3 天单纯 ICH 组(8.00±1.22、90.00±7.07)和再殖小胶质细胞组(7.13±1.27、83.75±9.92)小鼠mNSS评分(F=43.520,P＜0.05)及右侧转角百分比(F=39.180,P＜0.05)均显著高于假手术组;再殖小胶质细胞组mNSS评分及右侧转角百分比低于单纯ICH组,但差异无统计学意义(P＞0.05).ICH后第14天,再殖小胶质细胞组小鼠(3.25± 0.97、56.25±6.96)mNSS评分(F=109.400,P＜0.05)及右侧转角百分比(F=12.590,P＜0.05)明显低于单纯 ICH 组(4.75±1.39、68.75±7.81).且与单纯 ICH 组(术后第 3 天:2.63±0.35、18.57± 7.78;术后第14天:2.48±0.56、13.72±3.27)比较,再殖小胶质细胞组(术后第3天:1.47±0.31、4.46±4.56;术后第14天:1.20±0.02、4.57±1.83)小鼠血肿周围脑组织中IL-6、IL-1 β等促炎性细胞因子在 ICH 后第 3 天(IL-6:t=4.270,P＜0.05;IL-1β:t=2.710,P＜0.05)和第 14 天(IL-6:t=3.956,P＜0.05;IL-1β:t=4.232,P＜0.05)的表达量均显著低于单纯ICH组.结论 再殖小胶质细胞能改善小鼠ICH后长期神经功能缺损障碍,其机制可能是通过抑制神经炎症发挥作用.
Repopulating microglia improved neurological deficits in mice with hemorrhagic stroke
Objective To investigate the effect of repopulating microglia on the recovery of neuro-logical deficits in mice with intracerebral hemorrhage(ICH)and the underlying mechanism.Methods ICH model was prepared by stereotactic injection of collagenase Ⅳ into the striatum of C57BL/6J mice.The mice were divided into Sham group,ICH/original microglia group and ICH/repopulating microglia group.The modified neurological severity score(mNSS)and corner turning test were used to evaluate the neurological deficits of mice at 3rd and 14th day after ICH.Pro-inflammatory cytokines such as interleukin(IL)-6 and IL-1 β were analyzed using quantitative real time polymerase chain reaction.Results Compared to the sham group(0.38±0.48,50.00±8.88),the mNSS(F=43.520,P＜0.05)and corner turning rate(F=39.180,P＜0.05)in the ICH group(8.00±1.22,90.00±7.07)and the repopulating micro-glia group(7.13±1.27,83.75±9.92)were significantly increased on the 3rd day after ICH.Compared to the ICH group,the mNSS and corner turning rate of mice in the repopulating microglia group were lower,but the difference was not statistically significant(P＞0.05).On the 14th day after ICH,the mNSS(F=109.400,P＜0.05)and corner turning rate(F=12.590,P＜0.05)in the repopulating microglia group(3.25±0.97,56.25±6.96)were significantly lower than those in ICH group(4.75±1.39,68.75± 7.81,P＞0.05).In addition,compared to the ICH group(3rd d:2.63±0.35,18.57±7.78;14th d:2.48±0.56,13.72±3.27),the expression levels of pro-inflammatory cytokines in the perihematomal brain tissue of mice in the repopulating microglia group(3rd d:1.47±0.31,4.46±4.56;14th d:1.20±0.02,4.57±1.83)were significantly decreased on the 3rd d(IL-6:t=4.270,P＜0.05;IL-1β:t=2.710,P＜0.05)and 14th d(IL-6:t=3.956,P＜0.05;IL-1β:t=4.232,P＜0.05)after ICH.Conclusion Repopulating microglia improved the long-term neurological deficits after ICH in mice proba-bly through inhibiting neuroinflammation.
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