hsa_circ_0008039和miR-484在SK-N-SH细胞氧糖剥夺-复糖复氧损伤中的作用:与Fis1的关系

来源 :中华麻醉学杂志 | 被引量 : 0次 | 上传用户:tomlibu
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
目的:评价hsa_circ_0008039和miR-484在人神经母细胞瘤(SK-N-SH)细胞氧糖剥夺-复糖复氧(OGD/R)损伤中的作用及其与线粒体分裂蛋白1(Fis1)的关系。方法:体外培养SK-N-SH细胞至对数生长期,采用随机数字表法分为5组(n n=25):对照组(C组)、OGD/R组、hsa_circ_0008039 siRNA组(S组)、hsa_circ_0008039过表达组(E组)和hsa_circ_0008039 siRNA+miR-484抑制剂组(S+I组)。C组细胞在正常条件下培养;S组、E组和S+I组分别转染has_circ_0008039 siRNA、has_circ_0008039过表达载体、has_circ_0008039 siRNA和miR-484抑制剂后,氧糖剥夺12 h复糖复氧24 h制备OGD/R损伤模型。于复糖复氧24 h时采用CCK-8法检测细胞活力和LDH漏出量,采用RT-qPCR法检测hsa_circ_0008039、miR-484和Fis1 mRNA的表达,Western blot法检测Fis1表达;双荧光素酶报告验证hsa_circ_0008039与miR-484的靶向关系。n 结果:与C组比较,其余4组细胞活力降低,LDH漏出量增加,OGD/R组和E组hsa_circ_0008039和Fis1表达上调,miR-484表达下调,S组hsa_circ_0008039和Fis1表达下调,miR-484表达上调,S+I组hsa_circ_0008039和miR-484表达下调,Fis1表达上调(n P<0.05);与OGD/R组比较,E组和S+I组细胞活力降低,LDH漏出量增加,S组细胞活力升高,LDH漏出量减少,E组hsa_circ_0008039和Fis1表达上调,miR-484表达下调,S组hsa_circ_0008039和Fis1表达下调,miR-484表达上调,S+I组hsa_circ_0008039和miR-484表达下调,Fis1表达上调(n P<0.05);与S组比较,S+I组细胞活力降低,LDH漏出量增加,miR-484表达下调,Fis1表达上调(n P<0.01)。双荧光素酶报告实验表明:has_circ_0008039靶向结合miR-484。n 结论:hsa_circ_0008039靶向结合miR-484参与了SK-N-SH细胞氧糖剥夺-复糖复氧损伤的过程,与上调Fis1表达有关。“,”Objective:To evaluate the role of has_circ_0008039 and miR-484 in oxygen-glucose deprivation/reoxygenation (OGD/R) injury in SK-N-SH cells and the relationship with Fis1.Methods:SK-N-SH cells were cultured n in vitro to logarithmic growth stage and divided into 5 groups (n n=25 each) according to the random number table method: control group (group C), OGD/R group, has_circ_0008039 siRNA group (group S), hsa_circ_0008039 over-expression group (group E) and has_circ_0008039 siRNA plus miR-484 inhibitor group (group S+ I). Cells were cultured in normal condition in group C. In S, E and S+ I groups, after the cells were transfected with hsa_circ_0008039 siRNA, has_circ_0008039 over-expression vector, hsa_circ_0008039 siRNA and miR-484 inhibitor, the cells were subjected to oxygen-glucose deprivation for 12 h followed by 24 h restoration of On 2-glucose supply to develop the OGD/R model.At 24 h of restoration of On 2-glucose supply, the cell viability and amount of lactic dehydrogenase (LDH) released were measured using CCK-8 assay, the expression of hsa_circ_0008039, miR-484 and Fis1 mRNA was detected using real-time polymerase chain reaction, and the expression of Fis1 was detected by Western blot.A dual-fluorescein experimental report was used to verify the targeting relationship between hsa_circ_0008039 and miR-484.n Results:Compared with group C, the cell viability was significantly decreased, and the amount of LDH released was increased in the other 4 groups, the expression of hsa_circ_0008039 and Fis1 was significantly up-regulated, and the expression of miR-484 was down-regulated in OGD/R and E groups, the expression of hsa_circ_0008039 and Fis1 was significantly down-regulated, and miR-484 was up-regulated in group S, and the expression of hsa_circ_0008039 and miR-484 was significantly down-regulated, and the expression of Fis1 was up-regulated in group S+ I (n P<0.05). Compared with group OGD/R, the cell viability was significantly decreased, and the amount of LDH released was increased in E and S+ I groups, the cell viability was significantly increased, and the amount of LDH released was decreased in group S, the expression of hsa_circ_0008039 and Fis1 was significantly up-regulated, and the expression of miR-484 was down-regulated in group E, the expression of hsa_circ_0008039 and Fis1 was significantly down-regulated, and the expression of miR-484 was up-regulated in group S, and the expression of hsa_circ_0008039 and miR-484 was significantly down-regulated, and the expression of Fis1 was up-regulated in group S+ I (n P<0.05). Compared with group S, the cell viability was significantly decreased, the amount of LDH released was increased, the expression of miR-484 was down-regulated, and the expression of Fis1 was up-regulated in group S+ I (n P<0.01). The dual-fluorescein experimental report verified that miR-484 was the target of hsa_circ_0008039 which binded to miR-484 specifically.n Conclusions:has_circ_0008039 is involved in OGD/R injury in SK-N-SH cells by targetedly binding to miR-484, which is associated with up-regulation of Fis1 expression.
其他文献
肾间叶性软骨肉瘤是一种罕见的恶性软组织肿瘤。本文报道1例,患者因体检发现左肾肿瘤7d入院,行机器人辅助腹腔镜肾部分切除术,术后经病理和基因检测诊断为左肾间叶性软骨肉瘤。术后6个月复查,未见肿瘤局部复发和转移。“,”Renal mesenchymal chondrosarcoma is a rare malignant soft tissue tumor. This paper reports a patient who was admitted to hospital because of left re
目的:探讨整合耻骨上膀胱穿刺固定腺体法的经尿道前列腺等离子剜除术的可行性和安全性。方法:回顾性分析2020年1—6月广西医科大学第一附属医院收治的15例良性前列腺增生患者的临床资料。年龄(70.27±5.35)岁;术前血清前列腺特异性抗原(PSA)(3.03±1.37)ng/ml,前列腺总重量80.3(70.49,96.78)g;术前残余尿量80(55,108)ml,最大尿流率(Qn max)(6.13±2.25)ml/s;国际前列腺症状评分(IPSS)25 (22,27)分,生活质量评分(QO
目的:评价含十字形结构域蛋白3(JMJD3)在顺铂致急性肾损伤小鼠肾纤维化中的作用。方法:健康C57BL/6雄性小鼠48只,8~10周龄,体重20~30 g,采用随机数字表法分为4组(n n=12):对照组(CON组)、对照+ JMJD3抑制剂组(CON-A组)、顺铂组(CIS组)和顺铂+ JMJD3抑制剂组(CIS-A组)。CIS组和CIS-A组分别于第1和14天时腹腔注射顺铂15 mg/kg,制备急性肾损伤小鼠肾纤维化模型;第4天时分别腹腔注射JMJD3抑制剂GSKJ4 10 mg/kg和等
目的:探讨肠道嗜黏蛋白阿克曼菌(AKK)人群分布特征及其和肥胖关系的剂量效应,为该菌用于肥胖干预提供参考。方法:纳入2008年广东肠道微生物组计划中6 986名包含体重指数、腰围及常见混杂因素包括年龄、性别、舒张压、收缩压、空腹血糖、三酰甘油、总胆固醇、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇、尿酸的志愿者。根据细菌16S核糖体RNA(16S rRNA)基因测序信息,计算AKK属及其操作分类单元(OTUs)丰度;根据2002年中国肥胖工作组标准,诊断中心性或全身性肥胖。通过多变量n logisti
目的:探讨根治性膀胱切除患者行双侧输尿管单侧皮肤造口术的长期疗效。方法:回顾性分析2013年1月至2020年12月于上海市第十人民医院行输尿管皮肤造口术的104例膀胱癌患者的临床资料,其中66例根治术后行双侧输尿管单侧皮肤造口术(单侧组),38例行双侧输尿管双侧皮肤造口术(双侧组)。单侧组男53例,女13例;年龄(71.8±9.8)岁;体质指数(23.3±3.2)kg/mn 2。双侧组男33例,女5例;年龄(75.1±10.8)岁;体质指数(22.7±3.0)kg/mn 2;两组差异均
慢性前列腺炎/慢性盆腔疼痛综合征(CP/CPPS)是泌尿男科常见病之一,治疗后易复发。近年来,在传统药物疗效局限的情况下,体外冲击波治疗(ESWT)应用于CP/CPPS患者的安全性和有效性得到广泛证实。研究结果显示ESWT短期(≤ 12周)疗效令人满意,但缺乏长期(> 12周)的随访数据。此外,适应证不统一、作用机制未阐明等,限制了其进一步的临床推广。本文总结ESWT在CP/CPPS患者中应用的最新进展及其潜在作用机制。“,”Chronic prostatitis/chronic pelvic pain
通过尸检发现的前列腺癌也被称为前列腺潜伏癌。了解前列腺潜伏癌有助于全面认识前列腺癌,指导临床工作。现有的尸检研究提供了前列腺潜伏癌的患病率、Gleason评分、肿瘤体积、前列腺包膜外侵犯以及空间分布等重要信息。然而,不同的研究在研究时间、人群、研究方法以及结果呈现方面存在差异,使得整合这些结果较为困难。本文综述了目前前列腺潜伏癌的尸检研究结果,分析了尸检研究方法对结果的影响,提出了目前研究不足之处以及未来研究的方向。“,”Latent prostate cancer is defined as prost
畸胎瘤是一种少见的肿瘤,主要发生在卵巢、骶尾部、睾丸、中枢神经系统和纵隔,很少发生于肾脏,而原发于肾脏的成人非成熟性畸胎瘤极为罕见。本文报道1例成人左肾非成熟性畸胎瘤,行根治性肾切除术,病理检查示肿瘤组织内包含恶性上皮组织和少量脑组织。术后行化疗,随访26个月,死于全身转移导致的器官功能衰竭。“,”Teratoma is a rare tumor, mainly occurring in the ovary, sacrococcygeal region, testis, central nervous s