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.
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.
ObjectiveTo investigate the expression of miRNA-1 in denervated skeletal muscle at different periods, and to explore effects of passive movement on the expression of miRNA-1 and differentiation of myoblasts in denervation-induced skeletal muscle atrophy in rats. MethodsTwenty-seven Sprague Dawley rats, weighing (200±10) g, were randomly divided into sham-operated group (group A, n=3), denervated group (group B, n=12), and passive movement group (group C, n=12). After the right sciatic nerve was exposed and dissociated, the sciatic nerve of 1 cm in length was removed in groups B and C; resection was not performed in group A. At 1 day after operation, passive flexion and extension movement was performed on the right hind limb in group C. At 6 hours in group A and at 3, 7, 14, and 28 days in groups B and C, 3 rats were sacrificed to measure the wet weight ratio of gastrocnemius muscle, to observe the diameter of the gastrocnemius muscle cell and evaluate the muscle atrophy by HE staining; RT-PCR was used to detect the mRNA expression of miRNA-1 and myocyte differentiation factor (MyoD), and immunohistochemistry to determine the protein expression of MyoD. ResultsAtrophy in various degrees was observed in denervated gastrocnemius muscle of groups B and C. The muscle fiber arranged in disorder and the diameter of the muscle cells decreased gradually with the time, without normal structure and morphology. The wet weight ratio and the cell diameter of the gastrocnemius in groups B and C were significantly less than those in group A (P<0.05); the wet weight ratio at 7, 14, 28 days and the cell diameter at 7, 14 days of group B were significantly greater than those of group A (P<0.05). The expressions of miRNA-1 and MyoD mRNA gradually increased with time in groups B and C, but were significantly less than those of group A at each time point (P<0.05). At 7, 14, and 28 days after operation, the expressions of miRNA-1 and MyoD mRNA in group C were significantly higher than those in group B (P<0.05). Immunohistochemical staining showed positive expression of MyoD in groups A, B, and C at each time point, but higher expression was observed in groups B and C than group A; the expression increased with time in groups B and C, and it was significantly higher in group C than group B. The correlation analysis results showed that the overall change trend of miRNA-1 and MyoD had no relation with the gastrocnemius wet weight ratio at 3 and 7 days (P>0.05), and had positive correlation at 14 and 28 days (P<0.05); positive correlation was found between the relative expression of MyoD and miRNA-1 mRNA (P<0.05). ConclusionPassive movement can prevent amyotrophy by increasing the expression of miRNA-1 and promoting the differentiation of myoblasts.
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.
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.
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.
ObjectiveTo explore expression, clinical and biological significance of plasma miRNA-196a from patients with advanced gastric cancer.MethodsReal time quantitative RT-PCR (qRT-PCR) method was used to detect the miRNA-196a levels in tissues and plasma from 75 gastric cancer patients and 35 benign gastric lesions controls. Then clinic pathological correlations of plasma miRNA-196a in 75 gastric cancer patients were analyzed. Twenty-five gastric cancer patients were randomized selected from 75 patients, to compare plasma miRNA-196a levels between preoperation and postoperation. Meanwhile, the effect of miRNA-196a on the invasion ability of gastric cancer MGC-803 cell line was observed in vitro.ResultsThe levels of miRNA-196a in both plasma and tissues from 75 gastric cancer patients were significantly increased compared with 35 benign gastric lesions controls (P<0.000 1). Clinic pathological data of 75 gastric cancer patients showed that the expressions of miRNA-196a were significantly up-regulated in gastric cancer patients with serosal invasion (P<0.001), lymph node metastasis (P=0.004), distant metastasis (P<0.001) and late clinical stage (P<0.001). The expression of miRNA-196a in peripheral plasma of patients with gastric cancer was significantly down regulated after operation (P<0.000 1). In vitro, overexpression of miRNA-196a significantly increased the invasion ability of MGC-803 cells (P<0.05), whereas knockdown of endogenous miRNA-196a significantly inhibited the invasion ability of MGC-803 cells (P<0.05).ConclusionsThe expression of miRNA-196a is up-regulated not only in peripheral plasma of patients with gastric cancer, but also with the progression of gastric cancer (serosal invasion, lymph node metastasis and distant metastasis). The up-regulation of miRNA-196a expression in peripheral plasma is mainly due to the release of primary tumor tissue. miRNA-196a is expected to be a prognostic marker and a potential therapeutic target for advanced gastric cancer.
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.
ObjectiveTo explore the influence of miRNA-155/PU.1 signaling pathway blockade on bone marrow-derived dendritic cells (DCs) maturation and immune function of rat small intestinal transplantation. MethodsThe DCs were induced by adherent culture.The critical transcription factor gene PU.1 was designed and PU.1 siRNA was synthe-sized.The DCs were transfected by liposome transfection and a pair of PU.1 siRNA was screened according to the high silencing efficiency.The expressions of DCs surface markers CD80, CD86, and MHC-Ⅱamong three groups (PU.1 silent group, negative control group, and control group) were analyzed by flow cytometry.The IL-10 and IL-12p70 secretion levels in the supernatant were tested by ELISA method.The allogeneic T lymphocyte proliferation was tested by mixed lymphocyte reaction.The transfected cells were intravenously injected into the recipient rat on day 7 before intestinal transplantation.The survival conditions as well as pathological changes were observed in each group recipients. Results①The surface molecules CD80, CD86, and MHC-Ⅱin the PU.1 silent group were (27.0±5.6)%, (23.6±4.8)%, and (36.8±6.8)%, respectively; versus (74.0±9.4)%, (76.5±8.7)%, and (87.8±11.3)% in the negative control group, respectively, which were significantly lower in former and showing an in creased trend (P < 0.05).②Compared with the negative control group, IL-10 secretion level was significantly increased (P < 0.05), IL-12p70 secretion level significantly decreased (P < 0.05) in the PU.1 silent group.③The proliferation of T lymphocytes in the PU.1 silent group was significantly lower than that in the negative control group (P < 0.05).④When the transfected DCs were injected into the intestinal transplantation rats on day 7 before operation, the survival time was (14.3±3.3) d, (7.8±1.5) d, and (8.0±2.5) d in the PU.1 silent group, negative control group, and control group, respectively, which in the PU.1 silent group were significantly longer than that in the other two groups (P < 0.05), and the graft pathology showed that there were mild intestinal tissue damage, lymphocyte infiltration or villus edema in the PU.1 silent group. ConclusionmiRNA-155/PU.1 signaling pathway blockade could reduce DCs maturation and induce receptor-specific immune tolerance, which are proved both in vivo and in vitro.
Objective To explore the diagnostic value of miRNAs for pancreatic cancer. Methods PubMed, Scopus, Web of Science, CBM, CNKI, WanFang Data and VIP databases were retrieved from inception to December 31st 2015, to collect diagnostic accuracy studies about miRNAs for pancreatic cancer. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies by using the QUADAS-2 tool. Then meta-analysis was performed using MetaDiSc 1.4 and Stata 12.0 softwares. Results A total of 40 articles involving 109 studies were included. The results of meta-analysis showed that the pooled Sen, Spe, +LR, –LR and DOR were 0.81 (95%CI 0.80 to 0.82), 0.77 (95%CI 0.75 to 0.78), 3.15 (95%CI 2.78 to 3.58), 0.27 (95%CI 0.24 to 0.31) and 13.58 (95%CI 10.89 to 16.94), respectively. The AUC of SROC was 0.86 (95%CI 0.84 to 0.88). Subgroups analysis showed that: as to diagnostic accuracy, Caucasian was superior to Asian (AUC=0.89vs. 0.84); multiple-miRNAs profiling-based assays was superior to single miRNA assays (AUC=0.91vs. 0.84). Conclusion Current evidence suggests that miRNA has potential diagnostic value for pancreatic cancer, particularly using multiple miRNAs. Due to limited quality of the included studies, more high quality studies are needed to verify the above conclusion.