目的 探讨脑室周围白质软化（periventricular leukomalacia, PVL） 新生大鼠脑组织内源性白血病抑制因子（leukemia inhibitory factor, LIF）的表达变化。方法 采用左侧颈总动脉结扎并低氧（6% O2 4 h）处理建立 3 日龄 Wistar 大鼠缺氧缺血（HI）PVL 模型。HI 后 1 d、3 d、7 d、14 d 和 28 d 处死动物, 应用 Real-Time PCR 方法和 Western blot 方法分别检测脑组织 LIF mRNA 和蛋白的表达变化；应用免疫荧光双标染色方法检测
LIF 和胶质纤维酸性蛋白（glial fibrillary acidic protein, GFAP）的共表达情况。结果 HI 后 1、3、7 d PVL 组脑组织 LIF 蛋白表达量均高于对照组（均 P<0.01）；PVL 组脑组织 LIF 蛋白表达量于 HI 后 3 d 达到高峰，明显高于同组其他时间点（P<0.01），随后降低。LIF mRNA 的变化趋势与 LIF 蛋白一致。免疫荧光双标染色结果显示LIF 与 GFAP 存在共表达。结论 PVL 新生大鼠脑组织中星形胶质细胞 LIF mRNA 与蛋白的表达均呈先升高后降低的趋势；LIF 可能参与了 PVL 病变的修复过程。
Abstract
Objective To study the changes of endogenous leukemia inhibitory factor（LIF) in neonatal rats with periventricular leukomalacia（PVL). Methods A PVL model of 3-day-old Wistar rats was prepared by left carotid artery ligation followed by 6% oxygen for 4 hours. The rats were sacrificed at 1, 3, 7, 14 and 28 days of hypoxia ischemia（HI), and the brain tissues were sampled. Real-Time PCR and Western blot methods were applied to analyze the expression of LIF mRNA and protein. Double staining immunofluorescence was used to detect the co-expression of LIF and GFAP. Results At 1, 3 and 7 days of HI, LIF protein level in the PVL group was higher than in the control group（P<0.01). In the PVL group, the LIF protein level on the third day after HI reached a peak and was higher than the other time points（P<0.01). The change of LIF mRNA expression showed the same tendency with LIF protein. The double staining immunofluorescence showed a co-expression of LIF and GFAP. Conclusion LIF mRNA and LIF protein expression in astrocytes show a trend of initial increase followed by steady decline in neonatal rats with PVL, suggesting that endogenous LIF may participate in the repair of PVL.
关键词
白血病抑制因子 /
脑室周围白质软化 /
缺氧缺血 /
新生大鼠
Key words
Leukemia inhibitory factor /
Periventricular leukomalacia /
Hypoxia ischemia /
Neonatal rats
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