沉默Nogo-66受体的表达对宫内感染所致早产大鼠脑损伤的神经保护作用
发布时间：2019-06-02 16:11
【摘要】：目的探讨特异性si RNA沉默Nogo-66受体(Ng R)对宫内感染所致早产大鼠脑损伤修复的影响及作用机制。方法孕15 d Sprague-Dawley大鼠分别应用RU486和LPS诱导早产,随机选取RU486诱导的早产大鼠为对照组,将LPS诱导的宫内感染致脑损伤早产大鼠随机分为模型组、空载体组和Ng R-si RNA组,每组36只。对照组和模型组仅给予常规饲养,空载体组和Ng R-si RNA组均于出生后第1天(P1)经侧脑室1次性注入慢病毒空载体和Ng R-si RNA慢病毒载体后常规饲养。各组分别于P3、P7、P14时随机选取8只早产大鼠断头取脑。RT-PCR检测Ng R m RNA表达,Western blot测定活性Rho A蛋白表达,免疫荧光组化检测小胶质细胞活化程度和少突胶质前体细胞(OPCs)形态,苏木精-伊红染色观察脑组织病理学改变,P30时行动物行为学评分。结果 P3时,Ng R-si RNA组脑组织Ng R m RNA表达量、活性Rho A蛋白水平显著低于模型组和空载体组(P0.05);各组Ng R m RNA表达量与活性Rho A蛋白水平均呈正相关性(分别r=0.792、0.747、0.827、0.825,P0.05)。免疫荧光组化结果显示,Ng R-si RNA组P3时小胶质细胞CD11b荧光强度值较模型组和空载体组明显减弱(P0.05);各组O4抗体标记的OPCs细胞形态主要呈现三极突起形态。病理学结果显示,对照组脑室周围白质结构正常,染色清晰;模型组和空载体组白质结构疏松,纤维紊乱,可见软化灶;Ng R-si RNA组白质结构疏松,纤维紊乱相对较轻,胶质细胞增生不明显,无明显软化灶。行为学评分显示,Ng R-si RNA组的悬吊实验评分、活动总路程、平均速度和跨格次数大于模型组和空载体组,而斜坡实验时间及中心区活动时间和路程明显少于模型组和空载体组(P0.05),但同对照组比较差异均无统计学意义(P0.05)。结论 Ng R特异性si RNA可有效沉默宫内感染所致脑损伤早产大鼠Ng R基因表达,在脑损伤后的修复中具有显著神经保护作用。
[Abstract]:Objective to investigate the effect and mechanism of specific si RNA silencing Nogo-66 receptor (Ng R) on brain injury and repair induced by intrauterine infection in premature rats. Methods Sprague-Dawley rats on the 15th day of gestation were induced preterm delivery by RU486 and LPS, respectively. The premature rats induced by RU486 were randomly divided into model group, no-carrier group and Ng R-si RNA group. There are 36 rats in each group. The control group and the model group were only given routine feeding, while the empty carrier group and Ng R-si RNA group were fed with lentivirus empty vector and Ng R-si RNA lentivirus vector through the lateral ventricle on the first day after birth (P1). The brains of 8 premature rats were randomly selected at P3, P7 and P14. The expression of Ng R m RNA, Western blot was detected by RT-PCR. The expression of active Rho A protein was detected. The activation of microglia and the morphology of oligodendrocytes (OPCs) were detected by immunofluorescence histochemical method. The pathological changes of brain tissue were observed by hematoxylin-Ehong staining. The animal behavioral score was evaluated at P30. Results at P3, the expression of Ng R m RNA and the level of active Rho A protein in Ng R-si RNA group were significantly lower than those in model group and empty carrier group (P 0.05). The expression of Ng R m RNA was positively correlated with the level of active Rho A protein (r 鈮，
本文编号：2491251
[Abstract]:Objective to investigate the effect and mechanism of specific si RNA silencing Nogo-66 receptor (Ng R) on brain injury and repair induced by intrauterine infection in premature rats. Methods Sprague-Dawley rats on the 15th day of gestation were induced preterm delivery by RU486 and LPS, respectively. The premature rats induced by RU486 were randomly divided into model group, no-carrier group and Ng R-si RNA group. There are 36 rats in each group. The control group and the model group were only given routine feeding, while the empty carrier group and Ng R-si RNA group were fed with lentivirus empty vector and Ng R-si RNA lentivirus vector through the lateral ventricle on the first day after birth (P1). The brains of 8 premature rats were randomly selected at P3, P7 and P14. The expression of Ng R m RNA, Western blot was detected by RT-PCR. The expression of active Rho A protein was detected. The activation of microglia and the morphology of oligodendrocytes (OPCs) were detected by immunofluorescence histochemical method. The pathological changes of brain tissue were observed by hematoxylin-Ehong staining. The animal behavioral score was evaluated at P30. Results at P3, the expression of Ng R m RNA and the level of active Rho A protein in Ng R-si RNA group were significantly lower than those in model group and empty carrier group (P 0.05). The expression of Ng R m RNA was positively correlated with the level of active Rho A protein (r 鈮，
本文编号：2491251
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