长链非编码RNA TUG1调控miR-137参与局灶性脑缺血大鼠神经损伤的作用机制  

Regulatory mechanism of long non-coding RNA TUG1 on nerve injury in rats with by regulating miR-137

作　　者：张明菊 魏运兰 李建建 王炳旭 Zhang Mingju;Wei Yunlan;Li Jianjian(Department of Neurology,Luzhong Hospital,Pe-king University Medical College,Zibo Shandong255400;不详)

机构地区：[1]山东省淄博市北大医疗鲁中医院神经内科,255400 [2]山东省淄博市北大医疗鲁中医院儿科,255400

出　　处：《卒中与神经疾病》2021年第5期506-512,共7页Stroke and Nervous Diseases

摘　　要：目的探讨长链非编码RNA牛磺酸上调基因1(Long non-coding RNA taurine upregulation gene 1,LncRNA TUG1)调控微小RNA(MicroRNA,miR)-137参与局灶性脑缺血大鼠神经损伤的作用机制。方法将大鼠分为假手术组、模型组、过表达LncRNA TUG1组、敲低LncRNA TUG1组、空质粒(NC)组;Longa法评估大鼠神经功能;大鼠脑组织称重,计算脑组织含水量;脑组织2,3,5-三苯基氯化四氮唑(2,3,5-Triphenyltetrazolium chloride, TTC)染色观察脑梗死情况;脑组织尼氏染色观察海马CA1区神经细胞受损情况;比色法检测脑组织中半胱氨酰天冬氨酸特异性蛋白酶(Cysteing aspartate-spe-cific protease, Caspase)-3和Caspase-9活性;实时定量聚合酶链反应(Real-time quantitative polymerase chain reaction, RT-qPCR)检测脑组织中LncRNA TUG1和miR-137的表达水平;双荧光素酶法检测TUG1和miR-137靶向关系;蛋白质免疫印迹(Western blot, WB)检测脑组织中丝裂原活化蛋白激酶激活蛋白激酶2(Mitogen-activated protein kinase activated protein kinase 2,MK2)、肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)、白细胞介素-6(Interleukin-6,IL-6)的表达水平。结果与假手术组比较,模型组、NC组神经功能损伤评分、脑梗死占比、Caspase-3和Caspase-9活性、LncRNA TUG1以及MK2,TNF-α,IL-6表达水平均显著升高,miR-137表达水平显著降低(P<0.05);与模型组比较,过表达LncRNA TUG1组神经功能损伤评分、脑梗死占比、Caspase-3和Caspase-9活性、LncRNA TUG1以及MK2,TNF-α,IL-6表达水平均显著升高,miR-137表达水平显著降低(P<0.05),敲低LncRNA TUG1组神经功能损伤评分、脑梗死占比、Caspase-3和Caspase-9活性、LncRNA TUG1以及MK2,TNF-α,IL-6表达水平均显著降低,miR-137表达水平显著升高(P<0.05)。结论 LncRNA TUG1通过抑制miR-137表达来调控MK2介导的炎症反应,从而促进局灶性脑缺血大鼠神经损伤。Objective To explore the mechanism of long non-coding RNA TUG1(LncRNA TUG1) in nerve injury of rats with focal cerebral ischemia by regulating miR-137. Methods Rats were divided into sham operation group, model group, LncRNA TUG1 over expression group, LncRNA TUG1 knockdown group and NC(empty plasmid) group. Nerve function was evaluated by Longa method;the brain tissue was weighed and the brain water content was calculated;the infarction and the neurons in hippocampal CA1 area were observed by TTC staining and Nissl staining respectively;the activities of Caspase-3 and Caspase-9 in brain tissue were detected by colorimetry;the expression of LncRNA TUG1 and miR-137 in brain tissue were detected by real-time quantitative polymerase chain reaction(RT-qPCR);the targeting relationship between TUG1 and miR-137 was detected by dual luciferase assay;the expression of mitogen-activated protein kinase activated protein kinase 2(MK2), tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) in brain tissue were detected by Western blot. Results Compared with those in the sham operation group, the neurological impairment score, cerebral infarction rate, activities of Caspase-3 and Caspase-9, expression of LncRNA TUG1, MK2, TNF-α and IL-6 in the model group and NC group were significantly higher, the expression of miR-137 was significantly lower(P<0.05).Compared with those in the model group, the neurological impairment score, cerebral infarction rate, activities of Caspase-3 and Caspase-9, expression of LncRNA TUG1, MK2, TNF-α and IL-6 in the LncRNA TUG1 overexpression group were significantly higher, the expression of miR-137 was significantly lower(P<0.05), the neurological impairment score, cerebral infarction rate, activities of Caspase-3 and Caspase-9, expression of LncRNA TUG1, MK2, TNF-α and IL-6 in the LncRNA TUG1 knockdown group were significantly lower, the expression of miR-137 was significantly higher(P<0.05). Conclusion LncRNA TUG1 can regulate MK2-mediated inflammatory response by inhibiting the expression o

关 键 词：长链非编码RNA牛磺酸上调基因1 微小RNA-137 局灶性脑缺血 大鼠 丝裂原活化蛋白激酶激活蛋白激酶2 神经损伤 

分 类 号：R743.3[医药卫生—神经病学与精神病学]