卒中后外周免疫反应在老年雄性和雌性啮齿动物中受白血病抑制因子的差异调节。
The Poststroke Peripheral Immune Response Is Differentially Regulated by Leukemia Inhibitory Factor in Aged Male and Female Rodents.
机构信息
Department of Neurology, College of Medicine, University of Kentucky, Lexington, KY 740 S. Limestone, 40536, USA.
Department of Neuroscience, College of Medicine, University of Kentucky, Lexington, KY 800 Rose St., 40536, USA.
出版信息
Oxid Med Cell Longev. 2020 Dec 10;2020:8880244. doi: 10.1155/2020/8880244. eCollection 2020.
BACKGROUND
The goal of this study was to determine whether leukemia inhibitory factor (LIF) promotes anti-inflammatory activity after stroke in a sex-dependent manner.
METHODS
Aged (18-month-old) Sprague-Dawley rats of both sexes underwent sham surgery or permanent middle cerebral artery occlusion (MCAO). Animals received three doses of intravenous LIF (125 g/kg) or PBS at 6, 24, and 48 h before euthanization at 72 h. Spleen weights were measured immediately following euthanization. Western blot was used to measure protein levels of CCL8, CD11b, CXCL9, CXCL10, IL-12 p40, IL-3, and the LIF receptor (LIFR) in spleen tissue. ELISA was used to measure IL-1, IL-6, TNF, and IFN in spleen tissue. A Griess Assay was used to indirectly quantify NO levels via measurement of nitrite. Levels of cellular markers and inflammatory mediators were normalized to the baseline (sham) group from each sex. Statistical analysis was performed using two-way ANOVA and followed by Fisher's LSD post hoc test.
RESULTS
Aged female rats showed a significantly lower spleen weight after MCAO, but showed a significant increase in spleen size after LIF treatment. This effect was observed in aged male rats, but not to as great of an extent. CD11b levels were significantly higher in the spleens of MCAO+PBS males compared to their female counterparts, but there was no significant difference in CD11b levels between MCAO+LIF males and females. LIF significantly increased CXCL9 after LIF treatment in aged male and female rats. LIFR and IL-3 were upregulated after LIF treatment in aged females. Splenic nitrate increased after MCAO but decreased after LIF treatment in aged females. Splenic nitrate levels did not increase after MCAO but did increase after LIF treatment in aged males. The following cytokines/chemokines were not altered by sex or treatment: TNF, IL-6, IL-12 p40, CCL8, IFN, and CXCL10.
CONCLUSIONS
LIF treatment after permanent MCAO induces sex-dependent effects on the poststroke splenic response and the production of proinflammatory cytokines among aged rats.
背景
本研究旨在确定白血病抑制因子（LIF）是否以性别依赖的方式促进中风后的抗炎活性。
方法
年龄（18 个月）的雄性和雌性 Sprague-Dawley 大鼠接受假手术或永久性大脑中动脉闭塞（MCAO）。动物在安乐死后 72 小时前，于 6、24 和 48 小时接受三次静脉内 LIF（125μg/kg）或 PBS 注射。安乐死后立即测量脾脏重量。Western blot 用于测量脾脏组织中 CCL8、CD11b、CXCL9、CXCL10、IL-12 p40、IL-3 和 LIF 受体（LIFR）的蛋白水平。ELISA 用于测量脾脏组织中的 IL-1、IL-6、TNF 和 IFN。Griess 测定法通过测量亚硝酸盐间接定量 NO 水平。将细胞标志物和炎症介质的水平与每个性别组的基线（假手术）进行归一化。使用双向方差分析并随后进行 Fisher's LSD 事后检验进行统计分析。
结果
MCAO 后，老年雌性大鼠的脾脏重量明显降低，但 LIF 治疗后脾脏大小明显增加。这种作用在老年雄性大鼠中观察到，但程度较轻。MCAO+PBS 雄性大鼠的脾脏中 CD11b 水平明显高于雌性大鼠，但 MCAO+LIF 雄性和雌性大鼠之间的 CD11b 水平没有差异。LIF 显著增加了 LIF 治疗后老年雄性和雌性大鼠的 CXCL9。LIF 治疗后，老年雌性大鼠的 LIFR 和 IL-3 上调。MCAO 后脾脏硝酸盐增加，但 LIF 治疗后老年雌性大鼠的硝酸盐减少。MCAO 后脾脏硝酸盐水平没有增加，但 LIF 治疗后老年雄性大鼠的硝酸盐水平增加。以下细胞因子/趋化因子不受性别或治疗影响：TNF、IL-6、IL-12 p40、CCL8、IFN 和 CXCL10。
结论
永久性 MCAO 后 LIF 治疗会对老年大鼠中风后脾脏反应和促炎细胞因子的产生产生性别依赖性影响。
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