基于miR-223-3p研究淫羊藿苷治疗大鼠颅脑外伤后神经损伤的作用机制  

Mechanism of icariin on nerve injury in rats with craniocerebral trauma based on miR-223-3p

作　　者：房正华[1] 马宝林[1] 宋湖平[1] 代景伟[2] 孙晓欧[3] FANG Zheng-hua;MA Bao-lin;SONG Hu-ping;DAI Jing-wei;SUN Xiao-ou(Department of Neurosurgery,The First Affiliated Hospital of Xiamen University,Xiamen 361022,China;Department of Anesthesiology,The First Affiliated Hospital of Xiamen University,Xiamen 361022,China;Department of Neurosurgery,First Affiliated Hospital of Soochow University,Suzhou 215000,China)

机构地区：[1]厦门大学附属第一医院神经外科,福建厦门361022 [2]厦门大学附属第一医院麻醉科,福建厦门361022 [3]苏州大学附属第一医院神经外科,江苏苏州215000

出　　处：《中草药》2021年第19期5948-5955,共8页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81971171)。

摘　　要：目的基于miR-223-3p探讨淫羊藿苷治疗大鼠颅脑外伤神经损伤的作用机制。方法随机选取70只大鼠,建立局部脑挫裂伤模型,将建模成功的60只大鼠随机分为模型组、淫羊藿苷(30mg/kg)组、淫羊藿苷(30mg/kg)+阴性对照组和淫羊藿苷(30mg/kg)+miR-223-3pantagomir组,每组15只,另取15只大鼠作为对照组。给予相应药物进行干预,末次干预2 h后,对各组大鼠进行改良神经功能缺损评分(modified neurological severity scores,m NSS);测定各组大鼠脑组织含水量;采用ELISA法检测各组大鼠脑组织白细胞介素-18(interleukin-18,IL-18)和IL-1β水平;采用苏木素-伊红(HE)染色法观察各组大鼠脑组织病理变化;采用TUNEL染色法观察各组大鼠脑组织神经元凋亡情况;采用q RT-PCR法检测各组大鼠脑组织miR-223-3p和核苷酸结合寡聚化结构域样受体蛋白3(nucleotide-bindingoligomerizationdomain-likereceptor protein 3,NLRP3)m RNA表达情况;采用Western blotting法检测各组大鼠脑组织NLRP3、半胱氨酸蛋白酶-1(Caspase-1)、B淋巴细胞瘤2(B-cell lymphoma 2,Bcl-2)和Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)蛋白表达情况;采用双荧光素酶报告基因系统检测miR-223-3p对NLRP3的靶向性。结果与模型组比较,淫羊藿苷组大鼠m NSS显著降低(P<0.05),脑组织含水量显著降低(P<0.05),脑组织IL-18和IL-1β水平显著降低(P<0.05);脑组织损伤面积和红细胞数减少;神经元细胞凋亡指数显著降低(P<0.05);脑组织miR-223-3p mRNA表达水平升高,NLRP3 mRNA表达水平显著降低(P<0.05);脑组织NLRP3、Caspase-1和Bax蛋白表达水平降低,Bcl-2蛋白表达水平显著升高(P<0.05)。与淫羊藿苷组比较,淫羊藿苷+miR-223-3pantagomir可显著逆转上述指标(P<0.05)。双荧光素酶报告结果显示,miR-223-3p可靶向调控NLRP3。结论淫羊藿苷对大鼠颅脑外伤神经损伤具有改善作用,其作用机制可能与调控miR-223-3p从而靶向抑制NLRP3表达、抑制炎性反应、减少�Objective To explore the mechanism of icariin on nerve injury in rats with craniocerebral trauma based on mi R-223-3 p. Methods Seventy rats were randomly selected to establish a local brain contusion model. Sixty rats were successfully modeled and randomly divided into model group, icariin(30 mg/kg) group, icariin(30 mg/kg) + NC group and icariin(30 mg/kg) + mi R-223-3 p antagomir group, with fifteen rats in each group, and 15 rats were selected as control group. The corresponding drugs were given for intervention. 2 h after last intervention, rats in each group were subjected to record modified neurological severity scores(mNSS);Water content of brain tissue of rats in each group was measured;ELISA was used to detect interleukin-18(IL-18) and IL-1β levels in brain tissue of rats in each group;Hematoxylin-eosin(HE) staining method was used to observe the pathological changes of brain tissue of rats in each group;TUNEL staining method was used to observe neuron apoptosis in brain tissue of rats in each group;q RT-PCR was used to detect mi R-223-3 p and nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3) mRNA expressions in brain tissue of rats in each group;Western blotting was used to detect NLRP3, Caspase-1, B-cell lymphoma 2(Bcl-2) and Bcl-2 associated X protein(Bax) protein expressions in brain tissue of rats in each group;Dual luciferase reporter gene system was used to detect the targeting of miR-223-3 p to NLRP3. Results Compared with model group, mNSS of rats in icariin group was significantly reduced(P < 0.05), brain tissue water content was reduced(P < 0.05), IL-18 and IL-1β levels in brain tissue were reduced(P < 0.05);Brain tissue damage area and red blood cell count were decreased;Neuronal cell apoptosis index was decreased(P < 0.05);miR-223-3 p mRNA expression in brain tissue was increased, and NLRP3 m RNA expression was decreased(P < 0.05);NLRP3, Caspase-1 and Bax protein expressions in brain tissue were reduced, and Bcl-2 protein expression were increased(P < 0.05). Compared

关 键 词：淫羊藿苷 颅脑外伤 神经损伤 miR-223-3p NLRP3 炎性反应 

分 类 号：R285.5[医药卫生—中药学]