ObjectiveTo explore the effect of expression of miRNA-21 on bone marrow mesenchymal stem cells (BMSCs).MethodsIn this study, flow cytometry was used to identify the surface-associated antigens of BMSCs. The 10 μmol/L 5-azacytidine was used to induce BMSCs to differentiate to cardiomyocyte-like cells. Immunofluorescence was used to detect the expression of troponin I (cTnI). The samples were assigned to 3 groups: a blank group, a miRNA-21 mimic group, and a negative control (NC) group. The proliferation of BMSCs was detected by methyl thiazolylte-trazolium (MTT), the apoptosis of BMSCs was analyzed by flow cytometry. Western-blotting was used to identify the expression of cTnI and myod in the BMSCs.ResultsThe proliferation of BMSCs was increased, because of the over expression of miRNA-21. But the apoptotic rate of the BMSCs was slower in the miRNA-21 group, on account of the expression of miRNA-21 was higher than that in the NC group and the CK group. The expression of cTnI in the miRNA-21 group was higher than that in the NC group or the CK group.ConclusionThe results suggest that the up-regulation of miRNA-21 enhances proliferation of BMSCs, reduces the apoptosis of BMSCs. miRNA-21 promotes the differentiation of BMSCs, which may pave the way for the treatment directed toward restoring miRNA-21 function for myocardial ischemia.
Objective To detect expression of miR-483-5p in surem of patients with hepatocellular carcinoma (HCC) and investigate it’s clinical significance for diagnosis of HCC. Methods The rerum samples of 112 patients with HCC (HCC group), 85 patients with chronic viral hepatitis B (CHB group), and 56 healthy people for physical examination (healthy control group) were collected from January 2010 to January 2012 in the First Hospital of Lanzhou University. According to the results of preliminary chip detection of miRCURY LNATM miRNA, the real-time fluorescent quantitative PCR was adopted to quantitate the serum levels of miR-483-5p and miR-500a and the routine electrochemical method was used to detect the serum alpha fetoprotein (AFP) in every group. The receiver operating characteristic (ROC) curve was utilized to analyze the diagnostic values of serum miR-483-5p, miR-500a, and AFP for the HCC. Results The serum levels of miR-483-5p and miR-500a in the HCC group were significantly higher than those of the CHB and healthy control groups (both P<0.000 1), which had no significant differences between the CHB group and the healthy control group (P>0.05). The serum miR-483-5p level of the HCC patient decreased markedly at the postoperative 30 d (P<0.000 1) as compared with the preoperative level. The area under the ROC curve (AUC) of miR-483-5p, miR-500a, AFP, or miR-483-5p in combination with AFP for the diagnosis of the HCC was 0.74 (cutoff value=2.842, sensitivity=74% and specificity=66%), 0.66 (cutoff value=1.830, sensitivity=74% and specificity=51%), 0.81 (cutoff value=20 μg/L, sensitivity=78% and specificity=70%), and 0.92 (cutoff value=3.78, sensitivity=81% and specificity=83%), respectively. The AUC values of miR-483-5p in the diagnosis of the HCC patients with positive AFP (AFP>20 μg/L) and negative AFP (0–20 μg/L) were 0.78 and 0.83, respectively. Conclusions Serum miR-483-5p highly expresses in HCC, which has a certain accuracy in diagnosis of HCC, it combined with AFP could further increase its diagnostic value. Serum miR-483-5p might play an important supplemental role in diagnosis of HCC patient with negative AFP.
Mechanoresponsive microRNAs are a type of miRNAs which are sensitive or responsive to mechanical strain applied to them, their expression levels were changed after mechanical loading, thus affecting the expression levels of mRNA and proteins they regulated. Up to now, some mechanoresponsive miRNAs have been discovered in physiological or pathological tissues or organs. However, these discoveries are usually limited, and they are not able to guide clinical practice well. According to this situation, this paper summarizes research findings of mechanoresponsive miRNAs, and provides directions for clinical practice and further researches.
Objective To clarify that the vascular endothelial cell injury caused by obstructive sleep apnoea hypopnea syndrome (OSAHS) is partly mediated by miRNA-92a. Methods Serum miRNA-92a level was measured in patients who underwent polysomnography between January 2018 and December 2018. The correlation between miRNA-92a and OSAHS was analyzed. Meanwhile, endothelial cells were cultured in vitro, and morphological changes and JC-1 staining results of endothelial cells were observed after OSAHS serum stimulation, so as to further clarify the injury of endothelial cells. The changes of miRNA-92a target gene were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot to further clarify the mechanism of endothelial cell injury. Results Seventy-two patients received polysomnography, including 22 cases in the non-OSAHS group, 18 in the mild OSAHS group, 10 in the moderate OSAHS group, and 22 in the severe OSAHS group. Serum miRNA-92a level was significantly increased in the OSAHS patients, and it also increased with the aggravation of OSAHS severity. OSAHS serum significantly damaged endothelial cells. Endothelial cells were swollen, disordered arrangement, and unclear boundaries. JC-1 staining showed that green fluorescence was significantly enhanced compared with the control group. RT-PCR and Western blot showed that the expressions of Krüppel-like factor-2 (KLF-2), Krüppel-like factor-4 (KLF-4) and endothelial nitric oxide synthase (eNOS) were significantly decreased under OSAHS serum stimulation. Conclusion Serum miRNA-92a of OSAHS patients is significantly increased, and reduces the expression of target genes KLF-2, KLF-4 and eNOS, affects the mitochondrial function of endothelial cells, and injures endothelial cells.
Objective To screen pyroptosis-related miRNAs of acute aortic dissection (AAD) from the GEO database, and analyze and verify their functions. MethodsThe microarray data set based on the miRNA chip in the GEO database was downloaded, the differentially expressed miRNAs were screened, and the target genes were predicted by the miRWalk database. Pyroptosis-related genes (PRGs) were searched in the PubMed database with "pyroptosis" as the keyword, and the intersection of PRGs and differential miRNAs predicting target genes were taken as AAD PRGs by Venn diagram. GO and KEGG enrichment analyses were performed. CytoHubba was used to screen the critical AAD PRGs and then the AAD pyroptosis-related miRNAs were identified. Aortic tissues were collected from gender- and age-matched AAD patients and healthy people, and the critical PRGs and miRNAs were verified by Western blotting and RT-qPCR. ResultsA total of 46 AAD differentially expressed miRNAs were screened, and 49 AAD PRGs were obtained by Venn diagram. GO enrichment analysis showed that the genes played a vital role in apoptosis regulated by cysteine endopeptidases. KEGG analysis showed that the genes enriched in Salmonella infection, necroptosis, and Nod-like receptor signaling pathways. CytoHubba screened the critical AAD PRGs such as cysteine aspartase-1 (Caspase-1), tumor necrosis factor (IL)-1β, and tumor necrosis factor (TNF), then obtained 12 AAD pyroptosis-related miRNAs. Aortic tissues were collected from 6 AAD patients and 6 healthy people. There were 5 males and 1 females in the AAD group with an average age of 48.70±6.35 years, and 4 males and 2 females in the healty control group with an average age of 45.30±4.58 years. There was no statistical difference between the two groups in terms of gender, age, smoking history, hypertension, diabetes, or coronary heart disease (P>0.05). Western blotting and RT-qPCR results showed that Caspase-1 was up-regulated in the AAD patients' aortic tissues compared with the healthy aorta, and the corresponding miRNAs were miR-198, miR-3202, and miR-514b-5p, which were all down-regulated. Conclusion Through bioinformatics analysis and verification, the critical AAD PRGs are Caspase-1, IL-1β, and TNF, and Caspase-1 is up-regulated and 3 corresponding pyroptosis-related miRNAs are down-regulated, which provides new ideas for the molecular mechanism and targeted therapy of AAD cell pyroptosis.
ObjectiveThe aim of this meta-analysis and systematic review is to assess the effectiveness of microRNAs as a diagnostic tool for individuals with epilepsy. MethodsA systematic search of PubMed, EMBASE, the Cochrane Library, and Web of Science databases was performed to collect literature on miRNA diagnosis of epilepsy up to January 1, 2024. Two researchers independently screened and extracted the literature and resolved discrepancies by negotiation. The QUADAS-2 evaluation tool was used to assess the quality of the included studies. Statistical analysis was performed using Review Manager 5.4, Meta-Disc 1.4, and Stata 17.0. Results A total of 17 papers were included, including 942 patients with epilepsy and 932 healthy controls. miRNA in the diagnosis of epilepsy had a combined sensitivity of 0.76 [95%CI (0.71, 0.79)], combined specificity of 0.78 [95%CI (0.74, 0.82)], and area under the SROC curve of 0.84 [95%CI (0.80, 0.87)]. Subgroup analysis showed that miRNA had higher diagnostic value for temporal lobe epilepsy, especially medial temporal lobe epilepsy with hippocampal sclerosis (MTLE-HS). ConclusionThe study suggests that miRNA may be a promising tool for the diagnosis of epilepsy, especially temporal lobe epilepsy, but more high-quality studies are needed to support it.
ObjectiveTo systematically review the diagnostic value of miRNAs for Alzheimer’s disease (AD).MethodsPubMed, Web of Science, EMbase, The Cochrane Library, CNKI, WanFang Data, VIP, and CBM databases were electronically searched to collect diagnostic tests of miRNAs for AD from inception to October 31, 2020. Two researchers independently screened literature, extracted data, and assessed the risk of bias of the included studies. RevMan 5.3 and Stata 14.0 software were used for meta-analysis. ResultsA total of 22 studies involving 4 006 subjects were included. The meta-analysis results showed that the pooled sensitivity, specificity, positive and negative likelihood ratios, diagnostic odds ratio, and the areas under the working characteristic curve of miRNA in AD diagnosis were 0.83 (95%CI 0.79 to 0.87), 0.80 (95%CI 0.76 to 0.83), 4.07 (95%CI 3.37 to 4.92), 0.21 (95%CI 0.17 to 0.27), 19.20 (95%CI 12.96 to 28.48) and 0.88 (95%CI 0.85 to 0.90), respectively. ConclusionThe current evidence shows that miRNAs have a high diagnostic value for AD. However, because of the limited quality and quantity of the included studies, more high-quality studies are required to verify the above conclusion.
ObjectiveTo investigate the effect of Smad4 on the fibrosis of tendon derived fibroblasts (TDFs) induced by transforming growth factor β1(TGF-β1) by targeted regulation of miRNA219-5P (miR219-5P). MethodsThe tendons donated by the volunteers were harvested to isolate and culture TDFs. The 3rd generation cells were used for experiment. Chemically synthesized miR219-5P mimics, miR219-5P inhibitor, and negative control sequences were transfected into TDFs. The gene expression of miR219-5P in TDFs was detected by real-time PCR, and the protein expression of Smad4 in TDFs was detected by Western blot at 48 hours after transfection. The combining sites of miR219-5P and Smad4 in 3'UTR district were predicted by informatics software. Wild type and mutant type reporter gene expression vectors were constructed and then targeted verification was carried out by the luciferase reporter gene test. Transfected TDFs were then induced by TGF-β1. The proliferation activity of the cells were measured by the cell counting kit 8 after culturing for 24, 48, and 72 hours. The expressions of fibrosis related proteins in TDFs were detected by Western blot at 72 hours. ResultsAfter TDFs were transfected by miR219-5P mimics, miR219-5P expression was significantly up-regulated, but the expressions of Smad4 was decreased subsequently (P<0.05). Intracellular expression of miR219-5P was inhibited by miR219-5P mimics inhibitor, however, the protein expression of Smad4 was significantly increased (P<0.05). Luciferase reporter gene test showed that luciferase activities were significantly decreased in pGL3-WT-Smad4+mimics group, but were significantly increased in pGL3-WT-Smad4+inhibitor group when compared with pGL3-WT-Smad4 transfected group (P<0.05), but no significant difference was found between GL3-MT-Smad4+mimics and pGL3-MT-Smad4+inhibitor groups (P>0.05). Cell proliferation and the fibrosis related proteins were increased in TGF-β1 induced TDFs, however, decreased in TGF-β1 induced TDFs after transfected by miR219-5P inhibitor (P<0.01). ConclusionmiR219-5P can significantly inhibit fibrosis of TDFs induced by TGF-β1 by down-regulating Smad4 expression.
ObjectiveTo explore the dynamic expression changes of neuronal growth and differentiation-associated miR-124a and miR-9 in the process of epileptogenesis. MethodsEstablish the lithium-pilocarpine induced status epilepticus (SE) rat model. Animal behavior change induced by SE as well as in the period of chronic epilepsy was observed by naked-eye or video-recording. Major time points for the study were chosen at 1d, 7d, 14d and 28d post-SE, on which the post-SE rats were decapitated and their hippocampal specimens were obtained. Total RNA from each specimen was extracted and qPCR was exploited to detect miR-124a and miR-9 expression in the specimens. Statistical analysis was used to show the dynamic expressional changes of miR-124a and miR-9 in rat hippocampus at 1d, 7d, 14d and 28d post-SE during the process of epileptogenesis. ResultsCompared with normal rats, the expression level of miR-124a in rat hippocampus did not show a significant difference at 1d post-SE, but it had shown markedly differences at 7d, 14d and 28d post-SE(P < 0.05), with a declining trend. Compared with normal rats, the expression level of miR-9 had demonstrated significant differences at 1d, 7d, 14d and 28d post-SE(P < 0.05)with a generally increasing trend, although there was slight fluctuation of expressional up-regulation at 7d post-SE. ConclusionNeuronal growth and differentiation-associated miR-124a and miR-9 had shown dynamic changes of down-regulation or up-regulation in the process of epileptogenesis. It can be suspected that miR-124a and miR-9 take part in hippocampal neurogenesis post-SE and be involved in epileptogenesis process.
Objective To review the research progress of miRNA regulation in the differentiation of adipose-derived stem cells (ADSCs). Methods The recent literature associated with miRNAs and differentiation of ADSCs was reviewed. The regulatory mechanism was analyzed in detail and summarized. Results The results indicate that the expression of miRNAs changes during differentiation of ADSCs. In addition, miRNAs regulate the differentiation of ADSCs into adipocytes, osteoblasts, chondrocytes, neurons, and hepatocytes by regulating the signaling pathways involved in cell differentiation. Conclusion Through controlling the differentiation of ADSCs by miRNAs, the suitable seed cell for tissue engineering can be established. The review will provide a theoretical basis for molecular targeted therapy and stem cell therapy in clinic.