老年急性缺血性脑卒中患者血清miR-150-5p及miR-148b-3p的表达及其临床意义

目的探讨老年急性缺血性脑卒中(AIS)患者血清miR-150-5p及miR-148b-3p的表达水平及其临床意义。方法选取本院收治的178例老年AIS,按照美国国立卫生研究院卒中量表(NIHSS)评分分为:轻度组(52例,NIHSS评分5分)、中度组(83例,5分≤NIHSS评分≤20分),重度组(43例,NIHSS评分 20分)。另选择65例健康体检正常者作为对照组。采用实时荧光定量PCR检测各组血清miR-150-5p及miR-148b-3p表达水平。应用ROC曲线分析血清miR-150-5 p及miR-148 b-3 p表达水平对老年AIS诊断的价值。采用多因素logistic回归分析影响老年AIS的危险因素。结果 AIS组血清miR-125b-5p及miR-148b-3p水平明显低于对照组(P 0. 01)。重度组血清miR-125b-5p及miR-148 b-3 p水平均明显低于中度组和轻度组(P 0. 01)。血清miR-150-5 p及miR-148 b-3 p表达水平诊断老年AIS的最佳临界值分别为2. 82、1. 46,两项联合诊断老年AIS的AUC (95%CI)为0. 927 (0. 868～0. 991),其敏感度和特异度为93. 0%和86. 5%。多因素logistic回归分析显示,miR-150-5 p (OR=3. 107,95%CI:2. 194～6. 715)及miR-148 b-3 p (OR=2. 602,95%CI:1. 713～4. 350)低表达是老年AIS发生的独立危险因素。结论血清miR-150-5 p及miR-148 b-3 p表达水平在老年AIS患者中明显降低,是老年AIS发生的独立危险因素,有望作为老年AIS诊断的生物学标志物。     

miR-26a-5p alleviates CFA-induced chronic inflammatory hyperalgesia through Wnt5a/CaMKII/NFAT signaling in mice

Background

Inflammation often leads to the occurrence of chronic pain, and many miRNAs have been shown to play a key role in the development of inflammatory pain. However, whether miR-26a-5p relieves pain induced by inflammation and its possible mechanism are still unclear.

Methods

The complete Freund's adjuvant (CFA)-induced inflammatory pain mouse model was employed. Intrathecal or subcutaneous injection of miR-26a-5p agomir was performed after modeling to study its antinociceptive effect and the comparison of different administration methods. Bioinformatics analysis of miRNAs was performed to study the downstream mechanisms of miR-26a-5p. HE staining, RT-qPCR, Western blotting, and immunofluorescence were used for further validation.

Results

A single intrathecal and subcutaneous injection of miR-26a-5p both reversed mechanical hypersensitivity and thermal latency in the left hind paw of mice with CFA-induced inflammatory pain. HE staining and immunofluorescence studies found that both administrations of miR-26a-5p alleviated inflammation in the periphery and spinal cord. Bioinformatics analysis and dual-luciferase reporter gene analysis identified Wnt5a as a direct downstream target gene of miR-26a-5p. Wnt5a was mainly expressed in neurons and microglia in the spinal cord of mice with inflammatory pain. Intrathecal injection of miR-26a-5p could significantly reduce the expression level of Wnt5a and inhibit the downstream molecules of noncanonical Wnt signaling Camk2/NFAT, inhibiting the release of spinal cord inflammatory factors and alleviating the activation of microglia. In addition, miR-26a-5p could also inhibit lipopolysaccharide (LPS)-stimulated BV2 cell inflammation in vitro through a noncanonical Wnt signaling pathway.

Conclusions

miR-26a-5p is a promising therapy for CFA-induced inflammatory pain. Both intrathecal and subcutaneous injections provide relief for inflammatory pain. miR-26a-5p regulated noncanonical Wnt signaling to be involved in analgesia partly through antineuroinflammation, suggesting a pain-alleviating effect via noncanonical Wnt signaling pathway in the CFA-induced inflammatory pain model in vivo.     

血清miR-423-5p对生长激素腺瘤增殖的影响

目的 探讨生长激素腺瘤血清Micr0-RNA(miRNA)的表达情况,及miR-423-5p对生长激素腺瘤增殖的影响.方法 各检测6例生长激素腺瘤病人和正常人血清外泌体miRNA的表达情况,对比两者之间的差异.结果 外泌体miRNA表达谱显示:生长激素腺瘤病人和正常人血清之间有169个差异表达miRNA(P<0.05,...     

miR-1224-5p在胶质瘤中的表达及其对胶质瘤细胞增殖的影响

目的探讨miR-1224-5p在胶质瘤中的表达及其对人脑胶质瘤细胞增殖的影响。方法分析中国脑胶质瘤基因组计划(CGGA)数据库中miR-1224-5p在不同级别的胶质瘤组织中的表达数据,并采用原位杂交技术进行组织验证,运用CGGA数据分析miR-1224-5p的表达水平与胶质母细胞瘤患者临床预后的相关性。采用miR-1229-5p寡聚核苷酸转染胶质瘤U251及U87细胞,通过CCK8实验观察上调miR-1224-5p表达后对胶质瘤细胞增殖的影响。结果 miR-1224-5p在人脑胶质瘤组织中的表达随着病理分级的升高而降低。胶质母细胞瘤患者中低表达miR-1224-5p提示预后不良。上调miR-1224-5p表达可以抑制胶质瘤细胞的增殖。结论 miR-1224-5p与胶质瘤的级别和临床预后相关,miR-1224-5p可以抑制胶质瘤细胞的增殖能力。     

Neuron-derived exosomes with high miR-21-5p expression promoted polarization of M1 microglia in culture

BackgroundNeuroinflammation is a characteristic pathological change of acute neurological deficit and chronic traumatic encephalopathy (CTE) after traumatic brain injury (TBI). Microglia are the key cell involved in neuroinflammation and neuronal injury. The type of microglia polarization determines the direction of neuroinflammation. MiR-21-5p elevated in neurons and microglia after TBI in our previous research. In this study, we explore the influence of miR-21-5p for neuroinflammation by regulating microglia polarization.MethodsIn this study, PC12 and BV2 used to instead of neuron and microglia respectively. The co-cultured transwell system used to simulate interaction of PC12 and BV2 cells in vivo environment.ResultsWe found that PC12-derived exosomes with containing miR-21-5p were phagocytosed by microglia and induced microglia polarization, meanwhile, the expression of miR-21-5p was increased in M1 microglia cells. Polarization of M1 microglia aggravated the release of neuroinflammation factors, inhibited the neurite outgrowth, increased accumulation of P-tau and promoted the apoptosis of PC12 cells, which formed a model of cyclic cumulative damage. Simultaneously, we also got similar results in vivo experiments.ConclusionsPC12-derived exosomes with containing miR-21-5p is the essential of this cyclic cumulative damage model. Therefore, regulating the expression of miR-21-5p or the secretion of exosomes may be an important novel strategy for the treatment of neuroinflammation after TBI.     

Overexpression of miR-582-5p Inhibits the Apoptosis of Neuronal Cells after Cerebral Ischemic Stroke Through Regulating PAR-1/Rho/Rho Axis

Objective

The purpose of this study was to explore the role of miR-582-5p/proteinase-activated receptors type I (PAR-1)/Rho/Rho in neuronal cell apoptosis after cerebral ischemic stroke (CIS).

miR-384-5p ameliorates neuropathic pain by targeting SCN3A in a rat model of chronic constriction injury

ABSTRACT

Objective: To explore the potential regulation mechanisms of miR-384-5p in Neuropathic pain (NP).

Methods: Rat model of chronic constriction injury (CCI) was established to induce NP in vivo. NP levels were assessed using Withdrawal Threshold (PWT) and Paw Withdrawal Latency (PWL). qPCR and Western blotting were used to determine the relative expression of miR-384-5p and SCN3A. The inflammation response in spinal microglia cells was determined by ELISA assay. Immunofluorescence assay was used to demonstrate the co-localization of miR-384-5p with SCN3A in rat dorsal root ganglions (DRGs). The target genes of miR-384-5p were verified by dual-luciferase report assays.

Results: In the current study, the miR-384-5p expression level was significantly downregulated in CCI rats when comparing to the sham group. In addition, miR-384-5p agomir significantly repressed mechanical allodynia and heat hyperalgesia in CCI rats. Meanwhile, the current study indicated miR‐384‐5p could decrease inflammation progress in spinal microglia cells incubated in lipopolysaccharide. Consistently, overexpression of miR-384-5p obviously depressed inflammation cytokine levels in CCI rats. Dual-luciferase reporter assays indicated that SCN3A is a target gene of miR-384-5p.

Conclusion: miR-384-5p is a negative regulator in the development of neuropathic pain by regulating SCN3A, indicating that miR-384-5p might be a promising therapeutic target in the treatment of neuropathic pain.

Abbreviations: CCI: Chronic constriction injury; ZEB1: Zinc finger E box binding protein-1; MAPK6: Mitogen-activated protein kinase 6; COX-2: cyclooxygenase-2.     

重型颅脑损伤患者外周血miR-122-5p表达水平变化及其对近期预后的预测价值

目的 探讨重型颅脑损伤(sTBI)患者外周血miR-122-5p表达水平变化及其对近期预后的预测价值.方法 选择2016年3月-2019年8月本院收治的sTBI患者作为sTBI组,同期体检的健康志愿者作为对照组,检测外周血miR-122-5p的表达水平;随访sTBI组的近期预后并分为死亡患者和好转患者,分析预后的影响因...     

血清miR-122-5p水平与短暂性脑缺血发作病人预后的关 系

目的 探讨短暂性脑缺血发作（TIA）病人血清miR-122-5p水平变化及其在病人预后评估中的作用。方法 选取2013年1月至2016年1月收治的TIA病人150例,采集空腹静脉血检测血清miR-122-5p水平。根据入院ABCD2评分分为高危、中危、低危组。以发病90 d内发生脑梗死或死亡、与症状相关的大动脉狭窄≥50%或心源性栓子需要抗凝治疗为主要结局衡量终点事件。结果 150例中,低危组49例,中危组71例,高危组20例。150例中,41例（27.33%）出现终点事件,其中脑梗死10例,死亡1例,大动脉狭窄≥50%有21例,心源性血栓需抗凝治疗9例;其余109例（72.67%）无终点事件情况发生。血清miR-122-5p水平与病人ABCD2评分呈正相关（r=0.784,P<0.05）。发生终点事件TIA病人血清miR-122-5p明显高于无终点事件病人（P<0.05）,其中症状相关的大动脉狭窄≥50%病人、脑卒中/死亡病人血清miR-122-5p水平均明显增高（P<0.05）,而心源性栓子需抗凝治疗病人血清miR-122-5p水平无明显变化（P>0.05）。多因素logistic回归分析结果显示,血清miR-122-5p水平增高是TIA病人发生终点事件的独立危险因素（P<0.05）。结论 TIA病人血清miR-122-5p水平与ABCD2评分呈正相关;血清miR-122-5p水平增高提示TIA病人预后不良。     

miR-497-5p靶向叉头蛋白4基因对星形胶质细胞瘤细胞增殖、迁移和侵袭的影响

目的 探讨miR-497-5p靶向叉头蛋白4（FOXP4）基因对星形胶质细胞瘤细胞增殖、迁移和侵袭的影响。方法 采用qRT-PCR和Western blotting检测星形胶质瘤细胞（U87、U251和BT325）和正常星形胶质细胞HA1800中miR-497-5p和FOXP4的表达。采用MTT法和Transwell实验分析过表达miR-497-5p或沉默FOXP4对U251细胞增殖、迁移和侵袭的影响。双荧光素酶报告基因实验和Western blotting分析miR-497-5p和FOXP4靶向调控的关系。结果 3种星形胶质细胞瘤细胞中miR-497-5p的表达水平较HA1800细胞降低（P<0.05）,FOXP4的表达水平较HA1800细胞升高（P<0.05）。过表达miR-497-5p后U251细胞增殖、迁移和侵袭（均P<0.05）能力降低。沉默FOXP4后U251细胞增殖、迁移和侵袭（均P<0.05）能力降低。miR-497-5p靶向负性调控FOXP4表达。过表达FOXP4可部分逆转miR-497-5p对U251细胞增殖、迁移和侵袭的抑制作用。结论 miR-497-5p靶向下调FOXP4表达抑制星形胶质细胞瘤细胞的增殖、迁移和侵袭。     

大鼠颅脑损伤后脑组织miR-122-5p含量变化及其对神经功能的影响

目的 探讨大鼠颅脑损伤后脑组织miR-122-5p含量变化及其对神经功能的影响。方法 将40只成年SD大鼠随机分为假手术组,TBI模型组,乱序miRNA组,miR-122-5p模拟物组。采用Feeney法制备TBI大鼠模型;乱序miRNA组造模24 h后,立体定向注射乱序miRNA 5 μl（20 μmol/L）;miR-122-5p模拟物组造模24 h后,立体定向注射miR-122-5p模拟物组5 μl（20 μmol/L）。qRT-PCR法检测大鼠脑组织miR-122-5p的表达变化;免疫印迹法法检测p53蛋白表达水平;水迷宫法检测学习记忆功能;TUNEL法检测脑组织细胞凋亡情况;流式细胞术检测脑细胞线粒体膜电位的变化。结果 与假手术组大鼠比较,模型组和乱序miRNA组大鼠脑组织miR-122-5p表达明显减少,p53蛋白显著上调,脑组织细胞凋亡数明显增多,线粒体膜电位下降显著,水迷宫测试大鼠寻找隐形平台时间显著延长;与模型组大鼠比较,miR-122-5p模拟物组大鼠miR-122-5p表达明显增加,并伴随p53蛋白低表达,细胞凋亡水平下降,神经功能障碍得到明显逆转。结论 颅脑损伤后miR-122-5p的表达下调可能导致p53蛋白的表达升高,促进神经细胞凋亡。     

Development of a psychosocial functioning questionnaire for patients with major depressive disorder

BackgroundThe importance of psychosocial functioning in the assessment, diagnosis and treatment of major depressive disorder (MDD) is widely recognised. However, there is a lack of effective scales to assess psychosocial functioning in patients with MDD.AimsTo develop a professional questionnaire to evaluate the psychosocial functioning of patients with MDD.MethodsUsing a literature review, an open-ended questionnaire survey, and patient interviews, a theoretical model of psychosocial functioning was constructed, and an initial questionnaire was formed which included four dimensions. After two rounds of testing, using items analysis and exploratory factor analysis, a finalized questionnaire was created. There were 460 patients with MDD selected from six psychiatric hospitals for formal testing using a convenience sampling method. Forty patients were randomly selected and retested one week later to evaluate the test-retest reliability of the scale. The Patient Health Questionnaire-9 (PHQ-9), Short Form of Quality of Life Enjoyment and Satisfaction Questionnaire (Q-LES-Q-SF), Sheehan Disability Scale (SDS), and Dysfunctional Attitudes Scale (DAS) provided major standards to check the criterion validity of this questionnaire. Correlation analysis, confirmatory factor analysis, and internal consistency reliability testing were used to examine other psychometric characteristics of the finalized questionnaire.ResultsThe Psychosocial Functioning Questionnaire (PFQ) for patients with MDD included three dimensions: psychological cognitive functioning, subjective well-being, and social functioning, with a total of 18 items. The overall internal consistency reliability of the questionnaire was 0.957, and the test-retest reliability was 0.840. Confirmatory factor analysis showed that the model fitted well: Goodness of Fit Index (GFI)=0.888; Root Mean SquareError of Approximation (RMSEA)=0.085. The total score of PFQ was significantly correlated with the total score of PHQ-9, DAS, SDS and Q-LES-Q-SF (|r|=0.599–0.870, p<0.001).ConclusionsThe Psychosocial Functioning Questionnaire has good reliability and validity. It can be used to measure the psychosocial functioning of patients with MDD.     

血肿周围脑组织miR-340-5p、PDCD4表达水平与高血压性脑出血病人预后的关系系

目的 探讨高血压脑出血血肿周围脑组织miR-340-5p、程序性细胞死亡蛋白4（PDCD4）表达的相关性及临床意义。方法 收集2016年9月至2020年9月手术切除的高血压性脑出血血肿周围脑组织217例,选择同期病理科尸检获取的正常脑组织标本50例作为对照。PCR检测脑组织miR-340-5p、PDCD4 mRNA水平。发病90 d用改良Rankin量表评分评估预后,0~2分为预后良好,3~5分为预后不良。结果 217例中,预后良好148例,预后不良69例。血肿周围脑组织miR-340-5p水平明显低于对照组（P<0.05）,而PDCD4 mRNA水平明显高于对照组（P<0.05）。血肿周围脑组织miR-340-5p水平与PDCD4 mRNA水平呈明显负相关（r=-0.683,P<0.05）。预后不良组血肿周围脑组织miR-340-5p水平明显低于预后良好组（P<0.05）,而PDCD4 mRNA水平明显高于预后良好组（P<0.05）。多因素logistic回归分析显示,血肿周围脑组织miR-340-5p水平降低、PDCD4 mRNA水平增高是高血压性脑出血预后不良的独立危险因素（P<0.05）。ROC曲线分析显示,血肿周围脑组织miR-340-5p水平预测预后不良的最佳截断值为0.665,曲线下面积为0.808（95%置信区间0.733~0.882）,灵敏度、特异度分别为70.97%、71.05%;PDCD4 mRNA的最佳截断值为1.425,曲线下面积为0.834（95%置信区间0.775~0.894）,灵敏度、特异度分别为71.08%、68.35%。结论 高血压性脑出血后,血肿周围脑组织miR-340-5p表达减少,可能通过负向调控PDCD4参与脑损伤病理过程。血肿周围脑组织miR-340-5p、PDCD4 mRNA表达水平对发病90 d预后评估具有一定临床价值。     

脑胶质瘤病人血浆miR-9-3p的变化及其与病人临床预后的关系

目的 探讨脑胶质瘤病人血浆miR-9-3p的变化及临床意义。方法 选取2016年3月1日至2019年3月1日收治的脑胶质瘤42例,另选20例同期健康查体者为对照,用RT-PCR法检测血浆miR-9-3p水平。42例脑胶质瘤术后随访24～60个月,记录生存情况;以血浆miR-9-3p平均值为准,分为低表达和高表达。结果 脑胶质瘤病人血浆miR-9-3p水平明显降低（P<0.05）,多因素Cox分析显示,血浆miR-9-3P低表达是脑胶质瘤病人不良生存预后的独立危险因素（P<0.05）。生存曲线分析显示,miR-9-3p低表达脑胶质瘤病人中位总生存期较高表达病人明显缩短（P<0.05）。结论 人脑胶质瘤血浆miR-9-3p表达下调,与病人不良生存预后密切相关。     

miR-125a-5p靶向MEIS2基因对垂体瘤AtT20细胞增殖、凋亡的影响

目的探讨微小RNA-125a-5p(miR-125a-5p)是否可靶向调控MEIS2基因表达并分析其对垂体瘤AtT20细胞增殖及凋亡的影响。方法采用实时荧光定量PCR(qRT-PCR)检测AtT20细胞及小鼠正常垂体细胞(NPC)中miR-125a-5p及MEIS2 mRNA表达量。采用瞬时转染技术分别将miR-125a-5p mimic质粒、anti-miR-125a-5p抑制剂及si-MEIS2质粒转染至AtT20细胞。双荧光素酶报告基因检测miR-125a-5p与MEIS2的靶基因关系,同时共转染miR-125a-5p mimic与pcDNA-MEIS2验证miR-125a-5p是否通过靶向MEIS2表达发挥作用。MTT法检测各组细胞增殖能力;流式细胞术检测各组细胞凋亡率;蛋白免疫印迹(Western blotting)检测各组细胞MEIS2蛋白表达。结果 qRT-PCR检测结果显示,与NPC比较,miR-125a-5p在AtT20细胞中表达水平显著降低,MEIS2 mRNA表达水平显著升高;与miR-NC组比较,miR-125a-5p组miR-125a-5p表达水平显著升高。W...     

Up-regulation of Sirt1/miR-149-5p signaling may play a role in resveratrol induced protection against ischemia via p53 in rat brain

Micro-RNA(miRNA) are well studied small noncoding RNA, which plays a diverse role in the regulation of vital elements in cell survival and apoptosis. However, the functional significance of miRNAs after the pathogenesis of ischemic stroke remains unclear. The present study is designed to investigate the regulatory role of miR-149-5p on Sirtuin-1/p53 axis during ischemic-reperfusion-induced injury. Middle cerebral artery occlusion (MCAO) was performed by nylon monofilament for 60 min. Resveratrol was administered via intraperitoneal (IP) route, 30 min before the MCAO. Our study demonstrated that the miR-149-5p levels were markedly decreased at 24 h after ischemic-reperfusion (I/R) injury. Further, we observed decreased p53 protein expression and increased miR-149-5p activity on sirtuin1 (Sirt1) activation with resveratrol after 24 h following MCAO. Moreover, immunohistochemistry studies found that resveratrol treatment significantly decreased the immunoreactivity of p53 and caspase-3 on activation of Sirt1/miR149-5p axis. In conclusion, our findings suggest that miR-149-5p could play a regulatory role in neuronal cell death via Sirt1/p53 axis, which offers a new target for novel therapeutic interventions during acute ischemic stroke.