米诺环素对缺氧缺血脑损伤未成熟新生大鼠TLR4、NF-κBp65和TNF-α表达的影响  

Effects of Minocycline on expression of TLR4, NF-κBp65 and TNF-α in an developing rat model of hypoxic-ischemic brain damage

作　　者：喻婷[1] 李晓瑜[1] 邹晓萍[2] 皮荣标[3] 欧阳 余斯乐 车丽洪[5] 

机构地区：[1]中山大学附属第一医院儿科,广东广州510080 [2]广东省佛山市第一人民医院儿科,广东佛山528000 [3]中山大学药学院药理学与毒理学实验室,广东广州510006 [4]中山大学孙逸仙纪念医院,广东广州510120 [5]中山大学附属第一医院病理科,广东广州510080

出　　处：《解剖学研究》2013年第6期401-408,438,共9页Anatomy Research

摘　　要：目的观察米诺环素(Minocycline,MN)对缺氧缺血脑损伤(hypoxic-ischemic brain damage,HIBD)未成熟新生大鼠Toll样受体4(Toll-like receptor 4,TLR4)、核因子-κB(nuclear factor-kappa B,NF-κB)p65和TNF-α表达的影响,探索米诺环素脑保护作用机制。方法将160只生后2 d(P2)Sprague-Dawley(SD)新生大鼠随机分成正常对照组、假手术组、HIBD组、HIBD加MN组。通过结扎左侧颈总动脉及8%氮氧混合气缺氧4 h,制备未成熟新生大鼠缺氧缺血性脑损伤模型。HIBD加MN组大鼠缺氧后予腹腔注射1次MN 45 mg/kg,HIBD组予腹腔注射等剂量的无菌PBS(pH 7.4)。HI后24h、48 h、72 h取材,Western blotting检测TLR4、NF-κBp65和TNF-α蛋白表达,HI后72 h、4周行脑组织HE染色及病理评分,HI后4周行行为学检测。结果 HI后72 h、4周HIBD加MN组脑组织病理损伤较HIBD组减轻,HIBD加MN组半定量病理评分低于HIBD组,差异有统计学意义(P<0.05)。Western blotting显示HI后24、48和72 h HIBD加MN组TLR4、NF-κBp65和TNF-α的表达低于HIBD组,较正常组及假手术组升高。HI后4周,在悬吊试验及斜坡试验中,HIBD加MN组与正常组、假手术组差异无统计学意义(P>0.05)。旷场试验中,HIBD加MN组与正常组、假手术组对比,差异有统计学意义(P<0.05);与HIBD组比较,P=0.375,差异无统计学意义(P>0.05)。圆筒实验中HIBD加MN组左侧上肢触壁百分比较HIBD组降低(P<0.05),与正常组、假手术组差异无统计学意义(P>0.05);右侧触壁百分比的比较中,HIBD加MN组与正常组、假手术组、HIBD组差异无统计学意义(P>0.05)。结论米诺环素对缺氧缺血脑损伤近期及远期具有良好的保护作用,其对缺氧缺血脑损伤的保护作用机制可能与抑制TLR4-NF-κBp65-TNF-α信号途径的激活有关。Objective To explore the mechanism of neuroprotective effects of Minocycline, and observe the expression of TLR4, NF-κBp65 and TNF-a in immature rats with hypoxic-ischemic brain damage. Methods 160 Sprague-Dawley rats at the age of postnatal day 2 （P2） were randomly divided into 4 groups ： normal control group, sham operation group, HIBD group and HIBD＋MN group. HIBD group and HIBD＋MN group were subjected to the left common carotid artery ligation （CCAL） and exposure to 8% oxygen for 4 h. The HIBD＋MN group received a dose of minocycline （45 mg/kg） in PBS and the HIBD group received the same dose of sterile phosphate buffered saline （PBS） （pH 7.4） intraperitoneally immediately after the hypoxic exposure. 24, 48 and 72 hours after the HI insult, the expressions of TLR4, NF-κBp65 and TNF-a were detected by Western blotting. Brain tissues were collected on 72 hours and 4 weeks after HI for hematoxylin-eosin staining and histological scoring. Rats underwent behavioral tests on postnatal day 30. Results 72 hours and 4 weeks after the HI insult, the pathological changes were improved in HIBD＋MN group compared with the HIBD group. Compared with the HIBD group, histological scores in the left cerebral hemisphere of HIBD＋ MN group was significantly lower 72 hours and 4 weeks after HI （P〈0.05）. Western blotting showed that the expression of TLR4,NF-κBp65 and TNF-a protein in HIBD＋MN group were lower than that in HIBD group, and higher than normal group and sham group in 24, 48 and 72 hours after the HI insult. The outcomes of neurobehavioral test of HIBD group were abnormal.In the hanging test and inclined plane test, the scores of the HIBD＋MN group had no statistical significance compared with the normal group and sham group（P〉0.05 ）, but was higher than HIBD group. In the open field test, there was no significant statistical difference between the HIBD group and HIBD＋MN group （P=0.375）, but there was significant difference between these two groups and the normal gro

关 键 词：缺氧缺血 脑损伤 米诺环素 Toll样受体4 脑室周围白质软化 

分 类 号：R651.15[医药卫生—外科学]