Tofogliflozin attenuates renal lipid deposition and inflammation via PPARα upregulation mediated by miR-21a impairment in diet-induced steatohepatitic mice.

We previously demonstrated hepatic, cardiac, and skin inflammation in a high-fat diet-induced steatotic liver disease (SLD) model. However, the molecular mechanism in the kidneys in this model remains unclear. It has been recently reported that SGLT2 inhibitors improve chronic kidney disease (CKD). Therefore, we used this model to evaluate the effects of tofogliflozin on renal lipid metabolism and inflammation. Male 8-10-week-old C57Bl/6 mice were fed a high-fat/high-cholesterol/high-sucrose/bile acid (HF/HC/HS/BA) diet with 0.015% tofogliflozin (Tofo group) or an HF/HC/HS/BA diet alone (SLD group). After eight weeks, serum lipid profiles, histology, lipid content, and mRNA/microRNA and protein expression levels in the kidney were examined. The Tofo group showed significant reductions in body (26.9 ± 0.9 vs. 24.5 ± 1.0 g; p < 0.001) and kidney weight compared to those of the SLD group. Renal cholesterol (9.1 ± 1.6 vs. 7.5 ± 0.7 mg/g; p < 0.05) and non-esterified fatty acid (NEFA) (12.0 ± 3.0 vs. 8.4 ± 1.5 µEq/g; p < 0.01) were significantly decreased in the Tofo group. Transmission electron microscopy revealed the presence of fewer lipid droplets. mRNA sequencing analysis revealed that fatty acid metabolism-related genes were upregulated and NFκB signaling pathway-related genes were downregulated in the Tofo group. MicroRNA sequencing analysis indicated that miR-21a was downregulated and miR-204 was upregulated in the Tofo group. Notably, the expression of PPARα, which has been known to be negatively regulated by miR-21, was significantly increased, leading to enhancing ß-oxidation genes, Acox1 and Cpt1 in the Tofo group. Tofogliflozin decreased renal cholesterol and NEFA levels and improved inflammation through the regulation of PPARα and miR-21a.

The Association of Micro-RNA 21 and Hypertension: A Meta-Analysis.

Hypertension is a multifactorial, complex disease with high morbidity and mortality rates. Studies have found that micro-RNA 21 (miR-21) levels are significantly increased in patients with hypertension. However, other studies have reported opposite results. Therefore, the relationship between miR-21 expression and hypertension remains controversial. This meta-analysis was conducted to statistically evaluate the miR-21 levels of patients with hypertension. A literature research was conducted using Web of Science, Embase, PubMed, and CNKI. To search for titles or abstracts, 'hypertension' in combination with the terms 'miR-21,' 'microRNA-21,' or 'miRNA-21' were used as keywords. Standardized mean differences (SMD) with corresponding 95% confidence intervals (CIs) were determined from the results of the meta-analysis. In total, 12 articles were included in this meta-analysis, involving 546 cases and 436 controls. The results of the meta-analysis showed that miR-21 levels in patients with hypertension were significantly higher than those in the controls (SMD: 1.22; 95% CI [0.35, 2.09]). This meta-analysis is the first to evaluate miR-21 in patients with hypertension. MiR-21 may be a new target for the prediction and treatment of hypertension. Further high-quality studies are needed to better support the association between miR-21 and hypertension.

MicroRNA-21-5p agomir inhibits apoptosis of oligodendrocyte precursor cell and attenuates white matter injury in neonatal rats.

BACKGROUND AND RESEARCH QUESTION/HYPOTHESIS: Excessive oligodendrocyte precursor cell (OPC) apoptosis occurs during intrauterine infection-induced white matter injury (WMI) in premature infants, preventing excessive apoptosis of OPCs is one of the mechanisms protecting WMI. Micro-RNA-21-5p (miR-21-5p) mediating anti-apoptotic activity was observed in other diseases. Therefore, the aim of this study was to determine whether miR-21-5p protects against WMI by modulating phosphatase and tensin homologue deleted on chromosome 10/phosphatidylinositol-3-kinase/protein kinase B (PTEN/PI3K/Akt) signalling pathway. METHODS: A lipopolysaccharide (LPS)-induced neonatal Sprague-Dawley (SD) rat model of preterm WMI was established. To explore the effect of miR-21-5p on WMI, we intraventricularly injected miR-21-5p agomir and miR-21-5p antagomir to activate or inhibit endogenous miR-21-5p. Immunofluorescent labelling of myelin basic protein, immunohistochemical labelling of 2',3'-cyclic-nucleotide 3'-phosphodiesterase (CNPase), and terminal deoxynucleotidyl transferase dUTP nick end labelling assays were conducted to observe pathological white matter changes. The antibody of anti-oligodendrocyte marker 4 (O4) was used to specifically recognise OPCs. The expressions of miR-21-5p and PTEN mRNA in the brain were detected with quantitative real-time polymerase chain reaction (qRT-PCR). PTEN, Akt, and phosphorylated Akt (p-Akt) protein levels were assayed with western blotting, and apoptotic proteins associated with PI3K/Akt signalling were quantified. RESULTS: Intense white matter dysplasia and excessive OPC apoptosis were observed in the brains of rats with WMI. When the miR-21-5p agonist miR-21-5p agomir was used in the WMI group, apoptosis of OPCs was significantly reduced, and myelin maturation increased. MiR-21-5p agomir relieved WMI. MiR-21-5p agomir inhibited the mRNA and protein expression of PTEN, increased p-Akt phosphorylation, and decreased the expression and activation of related apoptotic proteins.On the other hand, the administration of miR-21-5p specific blocker, miR-21-5p antagomir, reduced the level of p-AKT, increased OPC apoptosis, and worsened WMI. INTERPRETATION: Our findings revealed that miR-21-5p agomir had anti-OPC over-apoptotic effects and enhanced myelin development in WMI by modulating the PTEN/Akt signalling pathway.

Relations of expressions of serum hypoxia inducible factor 2α and miR-21 with cerebral vasospasm in patients with aneurysmal subarachnoid hemorrhage after interventional embolization / 中华神经医学杂志

Objective:To investigate the relations of serum hypoxia inducible factor 2α (HIF-2α) and miR-21 expressions with cerebral vasospasm in patients with aneurysmal subarachnoid hemorrhage (aSAH) after interventional embolization.Methods:One hundred and seventy-four patients with aSAH underwent interventional embolization in our hospital from October 2017 to June 2019 were prospectively selected. DSA examination was performed 4 d after surgery, and severity of cerebral vasospasm was evaluated. Enzyme-linked immunosorbent assay was used to detect the level of serum HIF-2α and reverse transcription-PCR was employed to detect the serum miR-21 expression before and 3 and 7 d after interventional embolization. The clinical data and changes of serum HIF-2α and miR-21 expressions in patients with different degrees of cerebral vasospasm were compared. Correlation analysis was performed to analyze the relation of HIF-2α expression with miR-21 expression 3 and 7 d after interventional embolization. Receiver operating characteristics curve was used to analyze the diagnostic values of serum HIF-2α and miR-21 levels in cerebral vasospasm 3 d after interventional embolization.Results:There were 100 patients without vasospasm, and 20, 38 and 16 patients with mild, moderate and severe cerebral vasospasm, respectively. The serum levels of HIF-2α and miR-21 in patients with mild, moderate and severe cerebral vasospasm increased successively 3 and 7 d after interventional embolization, with significant differences ( P<0.05). Positive correlation was noted between expressions of HIF-2α and miR-21 in serum 3 and 7 d after interventional embolization ( P<0.05). Area under the curve (AUC) of HIF-2α in diagnosis of cerebral vasospasm was 0.748 ( 95%CI: 0.615-0.883, P=0.000) 3 d after interventional embolization. AUC of serum miR-21 level in diagnosis of cerebral vasospasm was 0.715 ( 95%CI: 0.590-0.842, P=0.000). AUC of serum HIF-2α combined with miR-21 in diagnosis of cerebral vasospasm was 0.893 ( 95%CI: 0.792-0.985, P=0.000). When diagnostic critical points of HIF-2α and miR-21 were 82.75 pg/mL and 1.15, the sensitivity, accuracy and negative predictive value of HIF-2α combined with miR-21 in the diagnosis of cerebral vasospasm were higher than those of HIF-2α or miR-21 alone. Conclusion:The expressions of serum HIF-2α and miR-21 in patients with aSAH after interventional embolization can effectively predict the occurrence of cerebral vasospasm, and may be involved in the occurrence and development of cerebral vasospasm.

Targeting MicroRNA-21 Suppresses Gastric Cancer Cell Proliferation and Migration via PTEN/Akt Signaling Axis.

MicroRNA plays a pivotal role in various human cancers, especially in human gastric cancer. In the present study, we evaluated the effect of microRNA-21 (miR-21) on the gastric cancer cell proliferation, migration, apoptosis and the related signaling cascades. Here, we showed that down-regulation of miR-21 markedly reduced gastric cancer cell proliferation (AGS and NCI-N87 cells) in a time dependent manner. Moreover, our findings revealed that silencing miR-21 dramatically blocked gastric cancer cell migration and movement, which might be related to down-regulation of vimentin expression. We also found that down-regulation of miR-21 promoted cell apoptosis and repressed cell cycle progression. Further investigation showed that down-regulation of miR-21 significantly increased phosphatase and tensin homolog (PTEN) protein expression level, but not transcription level (mRNA level), which in turn decreased Akt phosphorylation at Thr308 and Ser473. Collectively, our results uncover that miR-21 targets PTEN/Akt signaling pathway and regulates cell proliferation, migration and apoptosis in human gastric cancer cells. Our findings may provide a therapeutic target for treatment of human gastric cancer.

Study on expression and mutual regulation of serum IL-35 and miR-21 in septic patients / 国际检验医学杂志

Objective To detect the expression level and correlation of interleukin 35(IL-35) and micro RNA-21(miR-21) in septic patients and to explore the role of their mutual regulation in sepsis .Methods The serum was collected from the patients with sepsis and healthy controls .The expressions of serum IL-35 and miR-21 were detected by ELISA and real-time PCR ,and their correlation was analyzed ;mice were intraperito-neally injected by lipopolysaccharide(LPS) to establish the sepsis model ,then the mice were given anti-IL-35 p35 antibody ,IgG ,sepsis miR-21 mimics(agomir-21) and sepsis miR-21 negative control(agomir-21 NC) by in-traperitoneal injection ,after that ,the serum miR-21 expression level in each group was detected by real-time PCR and IL-35 ,TNF-αand IL-6 levels were detected by ELISA .Results The expression levels of serum IL-35 and miR-21 in sepsis patients were significantly increased compared with the healthy control group ,the difference was statistical significance (P<0 .05) ,moreover their expression levels showed the positive correla-tion (P<0 .05);after septic mice antagonizing to IL-35 ,the serum miR-21 expression was significantly in-creased ,meanwhile the expressions of inflammatory factor TNF-α and IL-6 were increased obviously ,the difference was statistical significance (P<0 .05) .After treating the septic mice by agomir-21 ,the serum IL-35 level was decreased significantly ,while the expression of inflammatory factors TNF-αand IL-6 were increased significantly ,the difference was statistical significance(P<0 .05) .Conclusion Serum IL35 and miR-21 in the patients with sepsis are jointly participate in the septic inflammatory response by mutual regulation .

Increased microRNA 21 expression contributes to arsenic induced skin lesions, skin cancers and respiratory distress in chronically exposed individuals.

More than 26 million people in West Bengal, India, are exposed to arsenic through drinking water, leading to several deleterious endpoints including precancerous and cancerous skin lesions and other non-dermatological health effects. Here, our aim was to identify whether miR21 is associated with such dermatological and non-dermatological health outcomes in chronically exposed humans. A total of 123 subjects from West Bengal were recruited for this study (45 exposed individuals with skin lesions, 38 exposed individuals without skin lesions and 40 unexposed individuals). The miR21 expression patterns in the lymphocytes were studied by quantitative realtime PCR and the effects on downstream targets were validated by Western blotting. Associations between the miR21 expression patterns and non-dermatological health effects were determined from epidemiological survey data. In vitro studies were done with low dose (0.05ppm) of chronic arsenic exposure to HaCaT cells for 15 passages. Interestingly, within the exposed group, the skin lesion individuals showed almost 4.5 fold up-regulation of miR21 compared to the no skin lesion group. The expression of the downstream targets of miR21 (PTEN and PDCD4) varied inversely, while the expression of pAKT and PI3K varied proportionately with its expression levels. Results of in vitro studies showed similar trends. Again miR21 was 2.03 fold up-regulated in the exposed individuals with respiratory diseases compared to the individuals without the same. This study for the first time shows that miR21 plays an important role in contributing to arsenic induced dermatological and non-dermatological health outcomes in an exposed population.

Updates of microRNA-21 in the occurrence and development of cholangiocarcinoma / 中华消化外科杂志

MicroRNA (miRNA) is an endogenous nonprotein-coding short-chain RNA,and it involves in the evolution of a variety of malignant tumors through targeted inhibition of mRNA translation or mRNA cleavage of gene expression regulation cancer or tumor suppressor genes,meanwhile,is also an important molecule in the occurrence and development processes of tumor.In the recent studies,miRNA has played an important role in the proliferation,apoptosis and invasion of cholangiocarcinoma cells,and it maybe become a new biomarker for the early diagnosis,assessment of prognosis and chemotherapy of cholangiocarcinoma.

Significance of micro RNA-21 expression in diffuse large B-cell lymphoma / 白血病·淋巴瘤

ObjectiveTo investigate the expression of miR-21 in diffuse large B cell lymphoma (DLBCL)and normal lymph tissues and its potential relevance with clinicopathological characteristics.MethodsThe expression levels of miR-21 in 50 primary DLBCL and 12 normal lymph node tissue specimens were examined by TaqMan real-time polymerase chain reaction.The expression of bcl-2 and p53 was detected by immunohistochemistry staining. ResultsThe expression of miR-21 was significantly higher in tumor tissues than that in normal tissues, in GCB subtypes higher than in non-GCB subtypes. And it was negatively correlated with bcl-2(P=0.020),while positively correlated with p53(P=0.022). Up-regulated miR-21 expression was low in three years of survival rate. ConclusionMiR-21 may indicate a more aggressive phenotype and serve as a molecular prognostic marker in DLBCL. High-expression of miR-21 is a key feature that is correlated with cell proliferation in DLBCL.miR-21 may have some guiding significance in prognosis.bcl-2,p53 is possibly one of the targets of miR-21 in DLBCL.