微小RNA-141-3p在脑出血患者血清中的表达及其作用机制  

MicroRNA-141-3p expression in serum of patients with cerebral hemorrhage and its mechanism

作　　者：李晓波[1] 夏鹰[1] 聂柳[1] 金虎[1] 李友俊[1] LI Xiao-bo;XIA Ying;NIE Liu;JIN Hu;LI You-jun(Department of Neurosurgery,Haikou People’s Hospital,Haikou 570208,China)

机构地区：[1]海口市人民医院神经外科,海口570208

出　　处：《解剖学报》2021年第4期506-511,共6页Acta Anatomica Sinica

基　　金：国家自然科学基金(81760234)。

摘　　要：目的分析微小RNA-141-3p(miR-141-3p)在脑出血(ICH)患者中的表达水平,并探讨miR-141-3p对大鼠脑出血损伤的作用及其机制。方法以40例脑出血患者和40例体检健康对照者为研究对象,采用Real-time PCR方法检测血清中miR-141-3p的含量。采用生物信息学预测miR-141-3p的靶标基因,双荧光素酶报告分析验证miR-141-3p对核苷酸结合寡聚结构域样受体蛋白3(NLRP3)的调控作用。在大鼠脑出血模型侧脑室注射miR-141-3p激动剂(agonist)或者激动剂对照(agonist NC),治疗7 d后进行神经功能评分。麻醉处死大鼠后取出脑组织,HE染色观察脑组织病理改变。干湿重法测定脑组织含水量,并采用Real-time PCR测定miR-141-3p和NLRP3的表达,通过Western blotting方法测定白细胞介素(IL)-1β、IL-6和IL-18的表达。结果与健康对照组(0.520±0.028)相比,脑出血患者血清中miR-141-3p的表达水平(0.068±0.038)显著下调(t=15.93,P<0.001),且与血肿周围水肿严重程度成负相关(r=-0.8948,-0.9434~-0.8087,P<0.001)。miR-141-3p靶向调控NLRP3基因表达。miR-141-3p的表达水平在miR-141-3p agonist组中明显高于agonist NC组(P<0.001),炎症小体NLRP3和炎症因子IL-1β、IL-6和IL-18表达水平明显低于agonist NC组(P<0.001)。与agonist NC组相比,agonist组miR-141-3p在ICH术后7 d脑水肿减轻,神经功能评分明显降低(P<0.001)。HE染色结果表明,ICH大鼠注射miR-141-3p激动剂后血肿周围水肿明显缩小。结论miR-141-3p通过靶向NLRP3降低炎症反应,减轻对脑出血后血肿周围脑组织的损伤。Objective To analyze the expression level of microRNA-141-3p(miR-141-3)in patients with intracerebral hemorrhage(ICH),and explore the effect and mechanism of miR-141-3p on cerebral hemorrhage injury in rats.Methods Forty patients with ICH and 40 healthy controls in total were enrolled in this study.The expression of miR-141-3p in peripheral blood serum was determined by the Real-time PCR method.The target relationship between miR-141-3p and nucleotide binding oligomerization domain-like receptor protein 3(NLRP3)3’UTR was confirmed by dual luciferase reporter assay.miR-141-3p agonist and agonist NC were injected into rats via the lateral ventricle,respectively.On day 7 after treatment,the neurological function score was evaluated,and then all rats were killed to obtain brain tissue.Brain water content was examined by the dried and wet mass.HE staining was conducted to observe the pathological changes of cerebral tissue.The m RNA expressions of NLRP3 and miR-141-3p were detected by Real-time PCR.The protein expression of interleukin(IL)-1β,IL-6 and IL-18 were detected by Western blotting analysis.Results The expression of miR-141-3p in serum of ICH patients was significantly down-regulated compared to healthy controls and negatively correlated with the severity of edema around the hematoma[(0.068±0.038)vs(0.520±0.028),t=15.93,P<0.001;r=-0.8948,-0.9434 to-0.8087,P<0.001].The result of luciferase reporter assay showed that miR-141-3p was related to the regulation of NLRP3 gene expression.The relative expression levels of miR-141-3p in agonist group were significantly higher than those in the agonist NC group(P<0.001),while the expression levels of NLRP3,IL-1β,IL-6 and IL-18 were significantly lower than those in the agonist NC group(P<0.001).Compared with the agonist NC group,the cerebral water content reduced significantly(P<0.001),and the neurological function score was significantly improved on the day 7 after treatment in agonist group(P<0.001).The result of HE staining showed that injection of miR-141-3p i

关 键 词：脑出血 NOD样受体蛋白3 微小RNA-141-3p 炎症反应 双荧光素酶报告分析 苏木素-伊红染色 人 大鼠 

分 类 号：R743.34[医药卫生—神经病学与精神病学]