褪黑素抑制早期蛛网膜下腔出血大鼠皮层小胶质细胞活化  

Melatonin inhibits the activation of cortical microglia in rats with early subarachnoid hemorrhage

作　　者：邹正 陈立刚 韩雨薇 郝广志 董玉书 Zou Zheng;Chen Ligang;Han Yuwei;Hao Guangzhi;Dong Yushu(Institute of Neuroscience,General Hospital of Northern Theater Command Base,Shenyang 110016,China)

机构地区：[1]中国人民解放军北部战区总医院神经外科,沈阳110016

出　　处：《神经解剖学杂志》2021年第3期310-314,共5页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(81671313,82071481);全军医学科技青年培育计划拔尖项目(17QNP053);中国博士后科学基金(2016M592951)。

摘　　要：目的:研究褪黑素对蛛网膜下腔出血大鼠(SAH)早期皮层小胶质细胞活化的影响及分子机制。方法:45只成年雄性SD大鼠分为假手术组(sham)、SAH组(SAH)和褪黑素治疗组(SAH+Melatonin)。利用注射自体血液的方法制备SAH大鼠模型,利用Western Blot检测大鼠皮层中小胶质细胞标记物CD11b的表达,利用real time RT-PCR检测大鼠皮层中诱导型一氧化氮合酶(iNOS)和环氧合酶-2(COX-2)mRNA的表达,利用酶联免疫吸附试验(ELISA)检测脑脊液中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)的表达,利用Western Blot检测大鼠皮层中toll样受体4(TLR4)和p38丝裂原活化蛋白激酶(p38 MAPK)的表达及磷酸化水平。结果:SAH模型大鼠皮层中CD11b的表达增加明显,iNOS和COX-2 mRNA表达上调,脑脊液中炎性因子IL-1β、IL-6、TNF-α水平增加;大脑皮层中TLR4表达增加,同时p38 MAPK磷酸化水平增加。褪黑素预处理可以显著逆转上述变化。结论:褪黑素预处理可以通过TLR4-p38 MAPK轴抑制早期SAH大鼠皮层小胶质细胞活化。Objective:To investigate the effect of melatonin on the activation of cortical microglia in early subarachnoid hemorrhage(SAH)rats and its molecular mechanism.Methods:Forty-five adult male SD rats were divided into sham operation group(sham),SAH group(SAH)and melatonin treatment group(SAH+Melatonin).The SAH rat model was prepared by injecting autologous blood.The protein expression of CD11b in the cortex was detected by Western Blot.The mRNA expression of inducible nitric oxide synthase(iNOS)and cyclooxygenase-2(COX-2)in the cortex was detected by real time RT-PCR.The levels of TNF-α,IL-1β,and IL-6 in the cerebrospinal fluid were detected by enzyme-linked immunosorbent assay(ELISA).The protein expression TLR4 and p38 MAPK in the cortex was detected by Western Blot.Results:The protein expression of CD11b was significantly increased in the cortex of SAH model rats.The mRNA expression of iNOS and COX-2 was up-regulated.The levels of TNF-α,IL-1β,and IL-6 in cerebrospinal fluid were increased.The expression of TLR4 and p38 MAPK phosphorylation in the cortex was increased.Pretreatment with melatonin can significantly reverse these changes.Conclusion:Preconditioning with melatonin can inhibit the activation of cortical microglia through TLR4-p38 MAPK axis in early SAH rats.

关 键 词：褪黑素 小胶质细胞 蛛网膜下腔出血 早期脑损伤 大鼠 

分 类 号：R743.35[医药卫生—神经病学与精神病学]