Inhibition of miR-429 improves neurological recovery of traumatic brain injury mice and attenuates microglial neuroinflammation

BackgroundNeuroinflammation is a common therapeutic target for traumatic brain injury (TBI) due to its contribution to delayed secondary cell death and has the potential to occur for years after the initial insult. Previous studies demonstrate that miR-429 is up-regulated in the brain lesions of TBI mice, while its role in regulating neuroinflammation and brain injury remains largely unknown.MethodThe expression of miR-429 in LPS-activated microglia and microglia in TBI model was detected by RT-PCR. The effects of miR-429 inhibitors on LPS-activated microglia in vitro as well as neurological recovery and post-traumatic neuroinflammatory response in TBI model mice were detected in vivo.ResultsLPS and TBI significantly induce the up-expression of miR-429, inflammatory cytokines, MAPK-p38 and phosphorylated NF-κB in microglia, which were all inhibited by miR-429 inhibitors. Meanwhile, miR-429 inhibitors also attenuated the neurological impairment in TBI mice. Bioinformatics analysis showed that miR-429 could target and inhibit the expression of dual specificity protein phosphatase 1 (DUSP1), thus inhibiting the expression of MAPK-p38 and phosphorylated NF-κB.ConclusionmiR-429 plays a pro-inflammatory role in activated microglia by targeting DUSP1 signaling pathway. Inhibiting miR-429 can attenuate the inflammatory response of microglia and TBI-mediated brain damage.     

Long noncoding RNA GAS5 promotes microglial inflammatory response in Parkinson's disease by regulating NLRP3 pathway through sponging miR-223-3p

Parkinson’s disease (PD) is the second most common neurodegenerative disorder. Neuroinflammation induced by microglia plays an important role in the pathogenesis of PD. Long noncoding RNA GAS5 was showed to have significant effects on regulating inflammatory response. Here, we aim to investigate the effects of GAS5 on the inflammatory response of PD, and the underlying mechanism. An in vivo model of PD was established in C57BL/6 mice by rotenone and an in vitro cell model was conducted on microglia by lipopolysaccharide (LPS). Our results indicated that GAS5 was upregulated in tissues in a mice model of PD and microglia activated by LPS. Gain- and loss- of functional experiments demonstrated that GAS5 promoted the inflammation of microglia in vitro. Besides, the knockdown of GAS5 repressed the PD progression in vivo. Mechanistically, GAS5 positively regulated the NLRP3 expression via competitively sponging miR-223-3p. Overall, our finding illuminates that GAS5 accelerates PD progression through targeting miR-223-3p/NLRP3 axis.     

circ-KEL在急性髓系白血病中的表达及其对白血病细胞的调控作用和机制

目的探讨circ-KEL在急性髓系白血病（AML）患者中的表达及其对AML细胞的调控作用和机制。方法收集116例AML患者以及40名健康者骨髓标本，分离骨髓单个核细胞，使用实时定量RT-PCR法检测circ-KEL的表达水平并分析其与AML患者临床特征之间的关系。通过生物信息学分析以及双荧光素酶报告基因实验等验证circ-KEL与miR-335-5p/LRG1的靶向关系。运用CCK-8、流式细胞术等方法检测circ-KEL在AML细胞中的生物学作用。结果circ-KEL在AML患者中的表达较正常人群明显增高（−7.117±1.831对−8.669±1.771，P<0.001），circ-KEL高表达患者总生存期（OS）明显短于低表达者（P＝0.037）。AML患者接受化疗后circ-KEL表达水平较初诊时下降。同时，circ-KEL可充当miR-335-5p的“海绵”，靶向调节LRG1。数据库分析显示miR-335-5p高表达患者预后好，且其与LRG1水平呈负相关。LRG1可促进AML细胞的增殖，抑制其凋亡，在AML患者中表达水平较健康人群也显著升高。circ-KEL可以通过miR-335-5p/LRG1轴发挥对白血病细胞的促增殖以及抑制凋亡作用。结论circ-KEL在AML患者中高表达，与AML患者预后高度相关，通过影响miR-335-5p/LRG1在AML疾病进展中发挥重要作用。     

Effects of propofol on cardiac function and miR-494 expression in rats with hepatic ischemia/reperfusion injury

ObjectiveThis study aimed to investigate the effects of propofol on cardiac function and miR-494 expression in rats with hepatic ischemia/reperfusion (I/R) injury.MethodsForty healthy adult male Sprague-Dawley rats were allocated to the sham operation group and three hepatic I/R injury groups. The I/R injury groups included I/R injury only (I/R group), treatment with propofol (propofol group), and treatment with propofol + overexpressed miR-494 (propofol+miR-494 group). Apoptosis of myocardial cells and changes in cardiac function indices, including left ventricular end-diastolic diameter, left ventricular end-systolic diameter, and left ventricular posterior wall thickness, as well as changes in miR-494, were monitored.ResultsThe apoptotic rate of myocardial cells in the I/R group was higher, cardiac function was deteriorated, and miR-494 levels were elevated compared with the sham group. The apoptotic rate was lower, cardiac function was improved, and miR-494 levels were suppressed in the propofol group compared with the I/R group. The apoptotic rate was higher, cardiac function was deteriorated, and miR-494 levels were elevated in the propofol+miR-494 group compared with the propofol group.ConclusionPropofol plays a vital role in preventing myocardial cell apoptosis and improvement of cardiac function by suppressing miR-494 in a hepatic I/R injury rat model.     

褪黑激素对体外培养神经元七氟醚损害的保护作用

目的 探讨褪黑激素（MLT）对体外培养神经元七氟醚（SEV）损伤的保护作用及其机制。方法 取SD大鼠乳鼠全脑皮层组织,分离皮质神经元进行体外培养;MLT预处理24 h后,4% SEV作用神经元;采用CCK-8法检测神经元存活率,流式细胞术检测神经元凋亡率,qPCR检测神经元miR-130a-3p和ROCK2 mRNA水平;免疫印迹法检测凋亡相关蛋白表达;应用在线预测软件TargetScan分析预测miR-130a-3p与ROCK2靶向关系并验证。结果 SEV明显降低神经元存活率以及miR-130a-3p和ROCK2 mRNA水平（P<0.05）,明显增加神经元凋亡率及凋亡相关蛋白cle-caspase3和cle-caspase9表达水平（P<0.05）;MLT明显抑制SEV的作用（P<0.05）。软件TargetScan分析显示,miR-130a-3p与ROCK2的3’非编码区第846~852碱基处存在结合位点,PCR和免疫印迹法检测显示,miR-130a-3p与ROCK2存在靶向关系。ROCK2过表达明显逆转MLT预处理的效果（P<0.05）。结论 MLT预处理明显改善SEV对体外培养的大鼠乳鼠皮质神经元的损害,机制可能是上调miR-130a,进而靶向抑制ROCK2表达。     

LncRNA 4344 promotes NLRP3-related neuroinflammation and cognitive impairment by targeting miR‐138-5p

ObjectiveCognitive impairment is a common neurological disease of which NLRP3-related neuroinflammation has been demonstrated to be an essential mediator. Previous studies have indicated that long non-coding RNAs (lncRNAs) are critical for the development of neurological disorders. However, the roles and functions of lncRNA 4344 in neuroinflammation during cognitive impairment are unknown and need to be further elucidated.MethodsLipopolysaccharide (LPS)-induced rat cognitive impairment and rat microglia (RM) cell inflammation models were established in vitro and in vivo. The Morris water maze test was used to evaluate the cognitive behavior of the rats. Gene expression was assessed using real-time quantitative polymerase chain reaction, and protein levels using enzyme-linked immunosorbent assay, or western blot analysis. The targeting relationship between lncRNA 4344, miR-138-5p, and NLRP3 was identified using bioinformatics analysis and a dual-luciferase reporter gene assay. Hematoxylin-Eosin and Nissl stainings, terminal deoxynucleotidyl transferase dUTP nick end labeling, or immunofluorescence staining assays were performed to detect pathological changes, neuronal apoptosis, or positive cells in hippocampal tissues, respectively.ResultsThe expression levels of lncRNA 4344 and NLRP3 were upregulated in the hippocampal tissues of LPS-treated rats and RM cells, and showed a strong positive correlation between each other. LncRNA 4344 overexpression further enhanced the expression of NLRP3 and its downstream genes (caspase-1, IL-1β, and IL-18), as well as neuronal apoptosis in LPS-stimulated RM cells, whereas lncRNA 4344 silencing attenuated the inflammatory injuries. Moreover, miR-138-5p was the direct target of lncRNA 4344 and was downregulated in the RM cell inflammation model. We also found that miR-138-5p directly reduced the expression of NLRP3 and its downstream genes. Subsequently, the results of the animal experiments showed that the lncRNA 4344/miR-138-5p/NLRP3 axis plays an essential role in regulating the cognitive behavior, pathological changes and apoptosis of hippocampal neurons, expression of inflammation-related factors (NLRP3, caspase-1, IL-1β, and IL-18), and microglial activation in LPS-induced cognitive impairment rats.ConclusionOur results demonstrated for the first time that lncRNA 4344 regulates NLRP3-related neuroinflammation and cognitive impairment by targeting miR-138-5p, providing a possible target for the treatment of diseases characterized by a cognitive deficit.     

CircAGFG1 acts as a sponge of miR-4306 to stimulate esophageal cancer progression by modulating MAPRE2 expression

ObjectiveThis work aims to determine the role of circular RNA (circRNA) AGFG1 and related molecular mechanism in esophageal squamous cell carcinoma (ESCC) cells.MethodsCircAGFG1 expression in ESCC cell lines was probed with qRT-PCR. ESCC cells were transfected/cotransfected with si-circAGFG1, pcDNA3.1-circAGFG1, si-Microtubule Associated Protein RP/EB Family Member 2 (MAPRE2), pcDNA3.1-circAGFG1 + miR-4306 mimic or pcDNA3.1-circAGFG1 + si-MAPRE2. The interactions between circAGFG1 and miR-4306 as well as miR-4306 and MAPRE2 were confirmed by dual-luciferase reporter assay. Cell proliferation, migration and invasion were detected by CCK-8, cell scratch and Transwell assays, respectively. Relative RNA expression levels of circAGFG1, miR-4306 and MAPRE2 in ESCC cells were measured by qRT-PCR. The protein level of MAPRE2 in ESCC cells was monitored by Western blot.ResultsCircAGFG1 was observably upregulated in ESCC cell lines. Besides, circAGFG1 silencing hindered ESCC cell development in vitro, and these effects were enhanced by miR-4306 overexpression or MAPRE2 silencing. Mechanistic analysis evidenced that circAGFG1 might act as a competitive endogenous RNA of miR-4306 to relieve the repressive effect of miR-4306 on its target MAPRE2.ConclusionCircAGFG1 facilitates ESCC progression via the miR-4306/MAPRE2 axis, and it may act as a possible biomarker for therapy and diagnosis in ESCC treatment.