scholarly journals MiR-384-3p aggravates propofol induced apoptosis in developing neurons by targeting Wnt3a

2021 ◽  
Vol 19 (12) ◽  
pp. 2505-2512
Author(s):  
Xiaoling Shi ◽  
Wei Jia
Keyword(s):  
Hippocampal Neurons ◽  
Neonatal Rat ◽  
Flow Cytometry Analysis ◽  
Chain Reaction ◽  
Expression Levels ◽  
Qrt Pcr ◽  
Diphenyltetrazolium Bromide ◽  
Polymerase Chain ◽  
Induced Apoptosis ◽  
Developing Neurons

Purpose: To evaluate the cytotoxicity of miR-384-3p toward propofol-treated neonatal rat hippocampal neurons and investigate its related molecular mechanism(s).Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to determine miR-384-3p expression levels in propofol-treated neonatal rat hippocampal neurons. Cell apoptosis and cell viability were evaluated by flow cytometry analysis and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, respectively. Bioinformatics and dual-luciferase reporterassay were applied together to forecast and verify the interaction between miR-384-3p and Wnt3a. Wnt3a expression in transfected neonatal rat hippocampal neurons was assessed by Western blot assay.Results: Propofol induced apoptosis in developing neurons by upregulating miR-384-3p expression levels. Knockdown of miR-384-3p reduced propofol-induced apoptosis in developing neurons. Subsequent experiments indicated that miR-384-3p directly regulated Wnt3a expression via coupling with the 3′-untranslated region of Wnt3a. Furthermore, upregulation of Wnt3a expression levels alleviated propofol-induced cytotoxicity promoted by miR-384-3p (p < 0.01).Conclusion: MiR-384-3p aggravates propofol-induced apoptosis in developing neurons by targeting Wnt3a. Keywords: MiR-384-3p, Wnt3a, Propofol, Apoptosis, Developing neurons

scholarly journals Seven-hydroxy-2-(4-hydroxy-3-methoxyphenyl)-chromen-4- one reduces atherogenic index and Nrf2 and GPx gene expressions in hyperlipidemic rats

2021 ◽  
Vol 18 (6) ◽  
pp. 1173-1177
Author(s):  
Prasetyastuti ◽  
Rahmah Dara Ayunda ◽  
Sunarti
Keyword(s):  
Density Lipoprotein ◽  
Atherogenic Index ◽  
Related Factor ◽  
Nuclear Factor ◽  
Chain Reaction ◽  
Gene Expressions ◽  
Expression Levels ◽  
Qrt Pcr ◽  
Polymerase Chain ◽  
Different Doses

Purpose: To investigate the effect of 7-hydroxy-2-(4-hydroxy-3-methoxyphenyl)-chromen-4-one isolated from mahogany (Swietenia macrophylla King) seeds on atherogenic index, expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and expression of the glutathione peroxidase (GPx) genes in hyperlipidemic rats. Methods: A total of 25 rats male aged 8 weeks and weighing an average of 200 g were used. They were divided into five groups as follows: (I) normal (N), (II) hyperlipidemic (HL), (III) hyperlipidemic rats treated with simvastatin (HL+SV), (IV and V) hyperlipidemic rats treated with 30 or 90mg, respectively, of 7-hydroxy-2-(4-hydroxy-3-methoxyphenyl)-chromen-4-one per 200 g body weight per day for 4 weeks. Atherogenic index (AI) was calculated from the levels of triglyceride (TG) and high-density lipoprotein (HDL) while Nrf2 and GPx gene expressions were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Results: Two different doses of 7-hydroxy-2-(4-hydroxy-3-methoxyphenyl)-chromen-4-one in hyperlipidemic rats significantly reduced their atherogenic index (p < 0.05). Nrf2 and GPx expression levels were lower than (p > 0.05) those of hyperlipidemic group. Conclusion: Seven-hydroxy-2-(4-hydroxy-3-methoxyphenyl)-chromen-4-one reduces the atherogenic index and expression levels of Nrf2 and GPx genes in hyperlipidemic rats. Thus, this compound has potential as an antihyperlipidemic agent

MicroRNA-582-5p Reduces Propofol-induced Apoptosis in Developing Neurons by Targeting ROCK1

2020 ◽  
Vol 17 (2) ◽  
pp. 140-146
Author(s):  
Zhongjie Zhang ◽  
Yan Xu ◽  
Songyuan Chi ◽  
Longji Cui
Keyword(s):  
Cell Viability ◽  
Hippocampal Neurons ◽  
Neurological Diseases ◽  
Regulatory Gene ◽  
Nerve Cells ◽  
Dependent Manner ◽  
Diphenyltetrazolium Bromide ◽  
Induced Apoptosis ◽  
Developing Neurons ◽  
Related Proteins

Background: Propofol is an intravenous drug commonly used in anesthesia procedures and intensive care in children. However, it also has neurotoxic effects on children. MicroRNA plays an important role in neurological diseases and neurotoxicity. Methods: In this study, primary rat hippocampal neurons were used to investigate the role of miR- 582-5p in propofol-induced neurotoxicity. Cell viability was monitored by 3-(4,5-dimethylthiazolyl)- 2,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, while the expression of proteins was monitored by real-time quantitation polymerase chain reaction (RT-qPCR) and western blot. TargetScan and double luciferase report assay were used to predict the targeting relationship between miR-582-5p and Rho-associated serine-threonine protein kinase 1 (ROCK1). Results: In the present study, the viability of neurons and the expression of miR-582-5p were decreased in a time-dependent manner after propofol treatment. Besides, miR-582-5p overexpression significantly reduced the toxicity of propofol on neuron cells but had no significant effect on normal nerve cells. In addition, miR-582-5p overexpression significantly reversed the expression of apoptosis-related proteins (cleaved caspase 3 and cleaved caspase 9) induced by propofol but had no significant effect in normal nerve cells. TargetScan and Dual-luciferase report assay revealed that ROCK1 was a targeted regulatory gene for miR-582-5p, and propofol treatment up-regulated ROCK1 expression by inhibiting miR-582-5p expression. Notably, miR-582-5p overexpression significantly increased cell viability, while ROCK1 overexpression reversed the effect of miR-582- 5p. Conclusion: Taken together, these findings suggest that miR-582-5p alleviated propofol-induced apoptosis of newborn rat neurons by inhibiting ROCK1.

scholarly journals Serum lnc34a is a potential prediction biomarker for bone metastasis in hepatocellular carcinoma patients

BMC Cancer ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Li Zhang ◽  
Hao Niu ◽  
Ping Yang ◽  
Jie Ma ◽  
Bao-Ying Yuan ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Bone Metastasis ◽  
Univariate Analysis ◽  
High Serum ◽  
Chain Reaction ◽  
Serum Samples ◽  
Early Screening ◽  
Expression Levels ◽  
Node Metastasis ◽  
Polymerase Chain

Abstract Background Early screening and intervention therapies are crucial to improve the prognosis of hepatocellular carcinoma (HCC) patients with bone metastasis. We aimed to identify serum lncRNA as a prediction biomarker in HCC bone metastasis. Methods The expression levels of lnc34a in serum samples from 157 HCC patients were detected by quantitative real-time polymerase chain reaction (PCR). Univariate analysis and multivariate analysis were performed to determine statistically significant variables. Results Expression levels of lnc34a in serum from HCC patients with bone metastasis were significantly higher than those without bone metastasis. The high expressions of lnc34a, vascular invasion and Barcelona Clinic Liver Cancer (BCLC) stage were associated with bone metastasis by analysis. Moreover, lnc34a expression was specifically associated with bone metastasis rather than lung or lymph node metastasis in HCC. Conclusions High serum lnc34a expression was a independent risk factor for developing bone metastasis in HCC.

scholarly journals Analysis and validation of a highly sensitive one-step nested quantitative real-time polymerase chain reaction assay for specific detection of severe acute respiratory syndrome coronavirus 2

2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Yang Zhang ◽  
Chunyang Dai ◽  
Huiyan Wang ◽  
Yong Gao ◽  
Tuantuan Li ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Quantitative Polymerase Chain Reaction ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Qrt Pcr ◽  
Highly Sensitive ◽  
One Step ◽  
Polymerase Chain

Abstract Background Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, is posing a serious threat to global public health. Reverse transcriptase real-time quantitative polymerase chain reaction (qRT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. Due to technical limitations, the reported positive rates of qRT-PCR assay of throat swab samples vary from 30 to 60%. Therefore, the evaluation of alternative strategies to overcome the limitations of qRT-PCR is required. A previous study reported that one-step nested (OSN)-qRT-PCR revealed better suitability for detecting SARS-CoV-2. However, information on the analytical performance of OSN-qRT-PCR is insufficient. Method In this study, we aimed to analyze OSN-qRT-PCR by comparing it with droplet digital PCR (ddPCR) and qRT-PCR by using a dilution series of SARS-CoV-2 pseudoviral RNA and a quality assessment panel. The clinical performance of OSN-qRT-PCR was also validated and compared with ddPCR and qRT-PCR using specimens from COVID-19 patients. Result The limit of detection (copies/ml) of qRT-PCR, ddPCR, and OSN-qRT-PCR were 520.1 (95% CI: 363.23–1145.69) for ORF1ab and 528.1 (95% CI: 347.7–1248.7) for N, 401.8 (95% CI: 284.8–938.3) for ORF1ab and 336.8 (95% CI: 244.6–792.5) for N, and 194.74 (95% CI: 139.7–430.9) for ORF1ab and 189.1 (95% CI: 130.9–433.9) for N, respectively. Of the 34 clinical samples from COVID-19 patients, the positive rates of OSN-qRT-PCR, ddPCR, and qRT-PCR were 82.35% (28/34), 67.65% (23/34), and 58.82% (20/34), respectively. Conclusion In conclusion, the highly sensitive and specific OSN-qRT-PCR assay is superior to ddPCR and qRT-PCR assays, showing great potential as a technique for detection of SARS-CoV-2 in patients with low viral loads.

Metformin Attenuates the Metabolic Disturbance and Depression-like Behaviors Induced by Corticosterone and Mediates the Glucose Metabolism Pathway

2021 ◽  
Author(s):  
Yong Hao ◽  
Yingpeng Tong ◽  
Yanhong Guo ◽  
Xiaoe Lang ◽  
Xinxin Huang ◽  
...  
Keyword(s):  
Glucose Metabolism ◽  
Tricarboxylic Acid Cycle ◽  
Antidepressant Activity ◽  
Tricarboxylic Acid ◽  
Serum Glucose ◽  
Metabolomic Analysis ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Metabolism Pathway ◽  
Polymerase Chain

Abstract Background Metabolism disturbances are common in patients with depression. The drug metformin has been reported to exhibit antidepressant activity. The purpose of this study was to investigate metabolism disturbances induced by corticosterone (CORT) and determine if metformin can reverse these effects and their accompanying depression-like behaviors. Methods Rats were exposed to corticosterone with or without metformin administration. Depression-like behaviors were tested. Gene expression was confirmed by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot analysis. In addition, the metabolites were quantified by LC-MS/MS analysis. Results Metformin attenuated the depression-like behaviors induced by CORT. Furthermore, metformin reversed disturbances in body weight, serum glucose, and triglyceride levels, as well as hepatic TG levels induced by CORT. Metformin normalized the alterations in the expression of glucose metabolism-related genes (PGC-1α, G6pc, Pepck, Gck, PYGL, Gys2, PKLR, GLUT4) and insulin resistance-related genes (AdipoR1, AdipoR2) in the muscles and livers of rats induced by CORT. Metabolomic analysis showed that metformin reversed the effects of CORT on 11 metabolites involved in the pathways of the tricarboxylic acid cycle, glycolysis, and gluconeogenesis (3-phospho-D-glycerate, β-D-fructose 6-phosphate, D-glucose 6-phosphate, and pyruvate). Conclusion Our findings suggest that metformin can attenuate metabolism disturbances and depression-like behaviors induced by CORT mediating the glucose metabolism pathway.

scholarly journals Downregulation of miR-383 reduces depression-like behavior through targeting Wnt family member 2 (Wnt2) in rats

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Shanshan Liu ◽  
Qing Liu ◽  
Yanjie Ju ◽  
Lei Liu
Keyword(s):  
Inflammatory Response ◽  
Family Member ◽  
Chronic Stress ◽  
Hippocampal Neurons ◽  
Luciferase Reporter ◽  
Mild Stress ◽  
Neural Injury ◽  
Expression Levels ◽  
Qrt Pcr

AbstractThis study aimed to evaluate the role of miR-383 in the regulation of Wnt-2 signaling in the rat model of chronic stress. The male SD rats with depressive-like behaviors were stimulated with chronic unpredictable mild stress (CUMS) including ice-water swimming for 5 min, food deprivation for 24 h, water deprivation for 24 h, stimulating tail for 1 min, turning night into day, shaking for 15 min (once/s), and wrap restraint (5 min/time) every day for 21 days. The expression levels of miRNAs were detected by qRT-PCR, and the expression levels of Wnt2, depression-impacted proteins (GFAP, BDNF, CREB), brain neurotransmitters (5-HT, NE, DA) and apoptosis-related proteins (Bax and Bcl-2) were evaluated by qRT-PCR and western blot. Bioinformatic analysis and luciferase reporter assay were performed to determine the relationship between miR-383 and Wnt2. Ethological analysis was evaluated by sugar preference test, refuge island test and open field tests. Rescue experiments including knockdown of miR-383, overexpression and silencing of Wnt2 were performed to determine the role of miR-383. High expression levels of miR-383 were observed in the hippocampus of rats submitted to CUMS model. Downregulation of miR-383 significantly inhibited the apoptosis and inflammatory response of hippocampal neurons, and increased the expression levels of GFAP, BDNF and CREB which were impacted in depression, as well as neurotransmitters, then attenuated neural injury in rats induced by CUMS. Furthermore, Wnt family member 2 (Wnt2) was identified as a target of miR-383, and silencing of Wnt2 obviously attenuated the protective effect of miR-383 inhibitor on the apoptosis and inflammatory response in hippocampal neurons, as well as neural injury in CUMS-induced rats. Downregulation of miR-383 ameliorated the behavioral and neurochemical changes induced by chronic stress in rats by directly targeting Wnt2, indicating that the miR-383/Wnt2 axis might be a potential therapeutic target for MDD.

Molecular Basis for Functional Differences of AMPA-Subtype Glutamate Receptors

Physiology ◽  
1996 ◽  
Vol 11 (2) ◽  
pp. 77-82 ◽  
Author(s):  
S Ozawa ◽  
J Rossier
Keyword(s):  
Polymerase Chain Reaction ◽  
Ampa Receptors ◽  
Molecular Basis ◽  
Hippocampal Neurons ◽  
Polymerase Chain Reaction Amplification ◽  
Inward Rectification ◽  
Chain Reaction ◽  
Functional Differences ◽  
Reaction Amplification ◽  
Polymerase Chain

To trace the molecular basis of functional properties of native a-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) receptors, we have coupled patch-clamp recordings and reverse transcription followed by polymerase chain reaction amplification. AMPA receptors lacking the GluR2 subunit in a population of hippocampal neurons exhibited a strong inward rectification and were highly permeable to Ca2+.

scholarly journals LncRNA PVT1 Promotes Hypoxia-Induced Cardiomyocyte Injury by Inhibiting miR-214-3p

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Chuanliang Liu ◽  
Jieqiong Zhang ◽  
Xuejie Lun ◽  
Lei Li
Keyword(s):  
Cell Viability ◽  
Cell Counting ◽  
Luciferase Reporter ◽  
H9c2 Cell ◽  
H9c2 Cells ◽  
Flow Cytometry Analysis ◽  
Chain Reaction ◽  
Cell Vitality ◽  
Gene Assay ◽  
Polymerase Chain

Objective. To explore the effect and related mechanism of LncRNA PVT1 on hypoxia-induced cardiomyocyte injury. Methods. PVT1RNA and miR-214-3p levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell vitality and apoptosis were, respectively, evaluated by Cell Counting Kit-8 (CCK-8) and flow cytometry analysis. Starbase and Dual luciferase reporter (DLR) gene assay was employed to validate the interaction between miR-214-3p and PVT1. Results. PVT1 was statistically upregulated, and miR-214-3p was statistically downregulated in hypoxia-induced H9c2 cells. The survival rate of H9c2 cells induced by hypoxia decreased statistically, while the apoptosis rate increased statistically ( P < 0.05 ). PVT1 knockdown upregulated the hypoxia-induced H9c2 cell viability and inhibited apoptosis. DLR assay verified the targeting relationship between PVT1 and miR-214-3p. In addition, miR-214-3p inhibitors reversed the viability of H9c2 cells with PVT1 knockout and promoted apoptosis. Conclusion. Silencing PVT1 can enhance the hypoxia-induced H9c2 cell viability and inhibit apoptosis, providing a potential target for the treatment of cardiovascular diseases.

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