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.     

Down-regulation of HCP5 inhibits cell proliferation,migration, and invasion through regulating EPHA7 by competitively binding miR-101 in osteosarcoma

Patients with osteosarcoma (OS) usually have poor overall survival because of frequent metastasis. Long non-coding RNAs (lncRNAs) have been reported to be associated with tumorigenesis and metastasis. In this study, we investigated the expression and roles of lncRNA human histocompatibility leukocyte antigen (HLA) complex P5 (HCP5) in OS, aiming to provide a novel molecular mechanism for OS. HCP5 was up-regulated both in OS tissues and cell lines and high expression of HCP5 was associated to low survival in OS patients. Down-regulation of HCP5 inhibited cell proliferation, migration, and invasion, suggesting its carcinogenic role in OS. miR-101 was targeted by HCP5 and its expression was decreased in OS. The inhibitor of miR-101 reversed the impact of HCP5 down-regulation on cell proliferation, apoptosis, and metastasis in OS. Ephrin receptor 7 (EPHA7) was proved to be a target of miR-101 and had ability to recover the effects of miR-101 inhibitor in OS. In conclusion, lncRNA HCP5 knockdown suppressed cell proliferation, migration, and invasion, and induced apoptosis through depleting the expression of EPHA7 by binding to miR-101, providing a potential therapeutic strategy of HCP5 in OS.     

MicroRNA-629-5p promotes osteosarcoma proliferation and migration by targeting caveolin 1

Osteosarcoma is a highly malignant tumor that occurs in the bone. Previous studies have shown that multiple microRNAs (miRNAs) regulate the development of osteosarcoma. This study aimed to explore the role of miR-629-5p and its target gene, caveolin 1 (CAV1), in osteosarcoma development. To analyze the expression of miR-629-5p and CAV1 mRNA in osteosarcoma tissues and cell lines, qRT-PCR analysis was performed. Dual-luciferase reporter experiments were subsequently performed to validate the relationship between CAV1 and miR-629-5p. CCK8 assay was used to measure osteosarcoma cell proliferation, and wound-healing assay was performed to study their migratory phenotype. Our findings revealed that miR-629-5p was overexpressed in osteosarcoma tissues and cells, and thereby enhanced cell proliferation and migration. Further, we validated that miR-629-5p targets CAV1 mRNA directly. CAV1 expression, which was negatively correlated with miR-629-5p expression, was found to be downregulated in osteosarcoma tissue samples. Moreover, our data showed that an increase in CAV1 level led to a decline in osteosarcoma cell proliferation and migration, which could be rescued by miR-629-5p upregulation. Overall, our study confirmed that miR-629-5p promoted osteosarcoma proliferation and migration by directly inhibiting CAV1.     

miRNA-125b Signaling Ameliorates Liver Injury Against Obstructive Jaundice-Induced Excessive Fibrosis in Experimental Rats

PurposeMultiple pathways are involved in inducing liver fibrosis, which can damage the integrity of liver. Among them, miR-125b has been found to exert an activating action on hepatic stellate cells. Endoplasmic reticulum stress and autophagy lead to liver disorders. Here, we evaluated the therapeutic influence of miR-125b on the endoplasmic reticulum function in injured livers submitted to bile duct ligation.Materials and MethodsFor inducing injury, bile duct ligation was done on miR-125b transgenic rats (miR-125b-Tg) in wild type rats. The rat T-6 cells received transfection of miR-125b mimic and Tunicamycin. Protein expressions were observed by western blot analysis.ResultsCompared to wild type rats, liver-injured rats showed significant impairment of liver function as assessed by the total bilirubin levels. The miR-125b-Tg rats showed decrease in activity of aspartate transaminase and alanine transaminase. Liver tissues of miR-125b-Tg rats showed weaker fibrotic matrix formation. Upregulation of miR-125b decreased the bile duct ligation-mediated hepatic disturbances for the expressions of endoplasmic reticulum kinase, inositol-requiring kinase 1alpha, sXBP1, CHOP, LC3, p62, ULK, and caspase-3/-8/-9. T-6 cells transfected with miR-125b mimic and treated with Tunicamycin caused decrease in levels of cleaved caspase-3, sXBP1, CHOP, and LC3. The miR-125b signaling showed protective effect on the liver tissues subjected to injury and fibrosis histopathology.ConclusionThis study demonstrates a novel insight into the miR125b-mediated stabilization of endoplasmic reticulum integrity, which slows the progression of injury-induced hepatic deterioration.     

miR-196a通过靶向NR6A1降低宫颈癌Hela细胞生存和运动能力

目的探究微小型RNA-196a(miR-196a)对宫颈癌Hela细胞生存和运动能力的影响和作用机制。方法RT-PCR检测正常宫颈上皮HcerEpic细胞和宫颈癌Hela细胞、CaSki细胞中miR-196a和核受体NR6A1的mRNA水平;阴性对照miR-NC和miR-196a inhibitor转染Hela细胞,分别记为NC组和miR-196a inhibitor组,免疫印迹检测NR6A1蛋白的表达,生物信息学和荧光素酶报告实验分析和验证miR-196a和NR6A1的靶向调控关系;阴性对照质粒pcDNA和过表达质粒pcDNA-NR6A1转染Hela细胞,分别记为pcDNA组和pcDNA-NR6A1组,免疫印迹检测NR6A1蛋白表达;将miR-196a inhibitor和pcDNA-NR6A1单独或联合转染Hela细胞,分别记为miR-196a inhibitor组、pcDNA-NR6A1组和inhibitor+NR6A1组,CCK8检测细胞增殖情况,流式细胞术检测细胞凋亡情况,Transwell检测细胞侵袭能力,划痕实验检测细胞迁移能力,免疫印迹检测Ki-67、Caspase-3、VEGF和MMP-9的表达。结果与HcerEpic细胞比较,Hela细胞和CaSki细胞中miR-196a和NR6A1 mRNA表达水平均较高(P<0.01),Hela细胞中miR-196a和NR6A1 mRNA表达变化最为显著(P<0.05);miR-196a inhibitor组Hela细胞中NR6A1蛋白表达显著降低(P<0.01),pcDNA-NR6A1组NR6A1的蛋白表达显著升高(P<0.01);miR-196a inhibitor能显著降低Hela细胞增殖倍数和Ki-67表达水平(P<0.05),同时还能提高Hela细胞凋亡率和Caspase-3表达(P<0.01);此外,miR-196a inhibitor还能减少侵袭细胞数,降低Hela细胞划痕闭合率并抑制VEGF和MMP-9的表达(P<0.01);NR6A1能显著减弱miR-196a inhibitor对Hela细胞增殖、凋亡、侵袭、迁移及对Ki-67、Casapse-3、VEGF和MMP-9表达的调控作用(P<0.01)。结论miR-196a inhibitor能通过靶向抑制NR6A1表达降低宫颈癌Hela细胞的生存和运动能力。