The application of iodinated contrast medium has become a risk factor for metformin-associated lactic acidosis (MALA), which leads to the accumulation of metformin in vivo is one of the principal reasons for MALA. However, the molecular mechanism of the adverse event is not yet clear. In this study, iohexol injection was used as a model drug. The contrast agent acute kidney injury rat model, in vivo rat pharmacokinetic study, kidney slices and HK-2 cells were performed to elucidate the pharmacokinetic molecular mechanism of accumulation of metformin caused by contrast-induced nephropathy (CIN). Plasma exposure of metformin was increased significantly in the CIN group compared with that in the normal and control groups. The AUC of metformin was from 2791 +/- 382 mu g min mL(-1) to 4784 +/- 767 mu g min mL(-1). The cumulative urinary excretion of metformin was also reduced in the CIN group. The uptake of metformin decreased in kidney slices in the CIN group. Compared with the normal and control groups, the blood lactate concentration was increased after intravenous administration of metformin in the CIN group followed a similar trend to the plasma concentrations of metformin. After treatment with contrast medium, the expression of OCT2 was reduced in rat kidney and HK-2 cells. These findings highlight that OCT2 deficiency was associated with increased lactate levels during metformin treatment caused by CIN.
Background: miR-208 family members have been considered as promising biomarkers in myocardial infarction (MI). Among which, miR-208a and miR-499 are reported to function as ischemic injury promoting miRNAs. This study aimed to explore the in vitro function of miR-208b in MI, which has not been widely studied. Methods: RT-qPCR was conducted to measure the expression changes of miR-208b in MI patients, MI mouse model and H9c2 cells stimulated by hypoxia. H9c2 cells were subjected to hypoxia before which miR-208b expression was altered by transfection. CCK-8, flow cytometry and Western blot were performed to detect cell survival. Besides, the regulatory relationship between miR-208b, Bax, and PI3K/AKT was tested by luciferase reporter, RT-qPCR and Western blot. Results: Serum levels of miR-208b in MI patients were significantly higher than those in the healthy controls. Also, miR-208b was up-regulated in mouse model and cell model of MI. Overexpression of miR-208b protected H9c2 cells against hypoxia-induced apoptosis, as the viability was increased, apoptosis rate was decreased, Bax and Cytochrome c were down-regulated, and Bcl-2 was up-regulated. Bax was a target gene of miR-208b. And miR-208b could not protect H9c2 cells when Bax was overexpressed. More interestingly, miR-208b activated PI3K/AKT pathway via targeting Bax, and the activated PI3K/AKT pathway could further repress Bax expression. Finally, blocking PI3K/AKT pathway by using LY294002 eliminated the myocardioprotective effects of miR208b. Conclusion: miR-208b is highly expressed during MI, and miR-208b protects H9c2 cells against hypoxia-induced apoptosis. miR-208b exerts myocardioprotective effect via targeting Bax and activating PI3K/AKT pathway.
Antioxidant therapy is considered as promising strategy for treating oxidative stress-induced cardiovascular disease. Bis (beta-elemene-13-yl) glutarate (BEG) is a novel beta-elemene derivative. Herein, we examined the antioxidant activity of BEG on human umbilical vein endothelial cells (HUVECs) after injury with hydrogen peroxide (H2O2) and investigated the mechanism involved. HUVECs were divided into the following groups: control group (untreated cells); treated groups (cells treated with 0.1, 1, 10 mu mol/L of BEG); positive control group (cells treated with 0.1 mM Vitamin E); model group (cells treated with 0.5 mM H2O2 alone). Cells were pre-incubated with or without BEG for 24 h and then incubated for a further 2 h with 0.5 mMH(2)O(2). Our results showed that BEG significantly reduced H2O2 induced loss in endothelial cell viability, reactive oxygen species (ROS) production, reduced lactate dehydrogenase (LDH) release, and malonyldialdehyde (MDA) level in a concentration-dependent manner. Also, BEG increased the cellular the superoxide dismutase (SOD) activity. Moreover, we found that H2O2 decreased Akt and eNOS phosphorylation, which perhaps, indirectly reduced nitric oxide (NO) production. These effects induced by H2O2, however, were reduced by pre-treatment with BEG. BEG effects were inhibited by a PI3K inhibitor (wortmannin) and eNOS inhibitor (L-NAME). In conclusion, the present study demonstrated that BEG has antioxidant activity. Furthermore, BEG reduced H2O2-induced endothelial cells injury by the involvement of antioxidation and PI3K/Akt/eNOS/NO signaling pathways.
Objectives: Pulmonary fibrosis is strongly correlated with inflammation factors, cytokine, and collagen secretion, whereby discoidin domain receptor 1 (DDR1) signaling plays an important role. EP300 is defined as an acetyltransferase that can acetylate histone and has been broadly studied in several chronic diseases, including cancer, inflammation and fibrosis. This study aimed to investigate the relationship between p300 and DDR1 in the pathological processes of pulmonary fibrosis. Materials and methods: Transcriptome analysis of single cell RNA-sequencing for idiopathic pulmonary fibrosis (IPF) bronchial epithelial cells demonstrated that both DDR1 and EP300 were up-regulated and involved in the regulation of autophagy, cellular response to organonitrogen compounds, and collagen metabolic pathways, respectively. The anti-fibrotic and anti-inflammation effects of Piml and DDR1 inhibitors in bleomycin-induced IPF murine models were estimated. Results: We discovered that overexpression of EP300 signaling induced MRCS human fibroblast cells that upregulated the expression of DDR1 and FN1; however, no effects on COL1 Al and DDR1 phosphorylation were observed. Mechanistically, TGF-beta 1 activated FN1, collagen, and DDR1 signaling could be reversed by the combination of p300 siRNA and DDR1 inhibitors. Moreover, the EP300 inhibitor SGC-CBP30 displayed synergistic effects with DDR1 inhibitors in pathogenic scores, airway goblet cell counts in bronchoalveolar lavage fluid (BALF), IL-4, IFN-gamma, FN1COL1 Al secretion and alpha-SMA, a marker of myofibroblast. Conclusions: The EP300 siRNA and inhibitors sensitized DDR1 inhibitors in our pulmonary fibrosis models in vitro and in vivo, implicating a combined inhibition of DDR1 with EP300 as potential therapies for IPF.
The endothelial-to-mesenchymal transition (EndMT) of endothelial cells contributes to the development of atherosclerosis. Oxidized low density lipoprotein (ox-LDL) is a highly risk factor for atherosclerosis. However, whether ox-LDL causes EndMT and the underlying mechanism are unclear. We report here that ox-LDL treatment is able to induce EndMT in human aortic endothelial cells (HAECs), and that the ox-LDL-induced EndMT is strictly dependent on the presence of its innate receptor, ox-LDL Receptor-1 (LOX-1). In addition, ox-LDL specifically upregulates EndMT transcriptional factor Snail, and knockdown of Snail completely attenuates ox-LDLinduced EndMT, indicating an essential role of Snail in mediating this effect. Mechanically, ox-LDL induces Snail stabilization by inhibiting its ubiquitination, which is in part attributed to inhibited GSK-3 beta activity. Hence, our findings suggest that inducing EndMT of aortic endothelial cells by ox-LDL might contribute to its detrimental role in promoting atherosclerosis development.
Osteoarthritis is the most common chronic condition of the joint disease. Chondrocyte is the sole cell type in joint tissues. Senescence of chondrocytes is known to contribute to the causation of osteoarthritis. Local inflammatory cytokines-caused chondrocytes senescence is proposed to be one of the molecular mechanisms of osteoarthritis. In this study, we show that the bile acid receptor GPBAR1 (TGR5), a G protein couples bile acid receptor, plays important roles in protecting chondrocytes from interleukin l beta (IL-1 beta)-caused senescence. TGR5 is fairly expressed in cultured chondrocytes. Its expression is reduced in isolated chondrocytes from osteoarthritis patients, and IL-1 beta treatment suppresses TGR5 expression. Activation of TGR5 by its synthetic agonist, INT-777, dramatically reduces senescence associated beta galactosidase activity by IL-1 beta. Mechanistically, the action of INT-777 ameliorates IL-1 beta-induced chondrocytes entry of G0/G1 arrest phase and exit of S and G2/M phases. INT-777 inhibits IL-1 beta-induced expression of p21, PAI-1, and K382 acetylation of p53 as well as reduction of Sirtl. The knockdown of TAGS abolished the protective role of INT-777 on these molecules. Collectively, our data indicates that activation of TGR5 is necessary for protection of IL-1 beta-induced chondrocytes senescence.
Endometriosis is one of the most common diseases in women. Inflammation and angiogenesis have been shown to be important in pathogenesis of endometriosis. Crocin is known as an anti-inflammatory, anti- proliferation substance. This study was designed to assess the potential effects of crocin on endometriosis. We established the mice model of endometriosis and administrated crocin to the mice. We monitored the endometriotic lesion growth, PCNA and VEGF expression in the lesion. We tested the serum levels of inflammatory cytokines in crocin-treated endometriosis mice. Finally we tested the effect of crocin on endothelial cell apoptosis and proliferation, and cytokine production in LPS-stimulated human monocyte. Crocin inhibited lesion growth in endometriosis mice and prevented PCNA and VEGF expression in the lesions. Crocin decreased the levels of inflammatory cytokines including INF-gamma, TNF-alpha, VEGF and IL-6 in serum. Crocin inhibited endothelial cells proliferation but did not cause apoptosis in endothelia cells. Crocin inhibited cytokine production in LPS-stimulated THP-1 cells in vitro. Crocin protected endometriosis by inhibiting endothelial cells proliferation and preventing inflammatory cytokines production.
Nowadays, neonatal sepsis has gradually become a global problem for its high incidence and increasing mortality. Previous studies have reported that miR-15a and miR-16 are two important modulators in neonatal sepsis. However, the upstream molecular mechanism of miR-15a/16 cluster is still mysterious. This study aims to explore a lncRNA can bind with miR-15a/16 in neonatal sepsis. Microarray analysis helped us found top ten lncRNAs which were downregulated in neonatal sepsis serum. Among these ten lncRNAs, SNHG16 was uncovered to significantly downregulated both miR-15a and miR-16. According to the result of subcellular fractionation assay, SNHG16 was mainly located in the cytoplasm of RAW264.7 cell, indicating the potential ceRNA role of SNHG16. Mechanism investigations revealed that SNHG16 could act as a ceRNA to upregulate TLR4 which is the target mRNA of miR-15a/16 cluster. At last, rescue assays demonstrated that SNHG16 reversed the effects of miR-15a/16 on LPS-induced inflammatory pathway. In summary, SNHG16 can act as a ceRNA to modulate miR-15a/16 cluster, thereby affecting LSP-induced inflammatory pathway which was downregulated by miR-15a/16 cluster.
Background: miR-516b, as a tumor suppressor in several tumors, its regulatory role in esophageal squamous cell carcinoma (ESCC) hasn't been previously reported. Objective: This study was to investigate the potential role of miR-516b in ESCC. Methods: miR-516b expression was measured in ESCC tumor specimens and matched adjacent non-cancerous tissues from 80 ESCC patients. The association between miR-516b and clinicopathological features of these patients was analyzed. The effect of miR-516b was evaluated by cell proliferation, migration, invasion and apoptosis assays in ESCC cell line EC9706 and TE-9. The role of miR-516b in vivo was further studied by constructing ESCC xenograft mice model. The direct target of miR-516b was predicted by public miRNA database and confirmed by luciferase reporter assay. The regulation of miR-516b on the target gene was further confirmed in vitro and in vivo. The expressions of proteins related to cell cycle and apoptosis were analyzed by western blot analysis, and cell migration and invasion were assessed by transwell assays. Results: miR-516b expression was reduced in ESCC tissues and cells, and correlated with advanced TNM stage, depth of invasion, lymphatic metastasis and poorer overall survival in ESCC patients. miR-516b was upregulated by miR-516b mimics repressing cell proliferation, and inducing G1 cell cycle arrest and apoptosis. miR-516b upregulation also suppressed the growth of ESCC xenograft tumor in nude mice and the invasion of ESCC cells via regulating the epithelial-mesenchymal transition pathway. CCNG1 was identified as a direct downstream target of miR-516b. Conclusion: The results demonstrated miR-516b functions as a tumor suppressor by directly modulating CCNG1 expression in ESCC cells, and may be a novel therapeutic and prognostic biomarker for ESCC.
Bronchopneumonia is a common multiple infection disease under 2 years old. Luteolin is a natural flavonoid widely distributed in plants with anti-inflammatory effect. This study aimed to explore the effects of luteolin on lipopolysaccharide (LPS)-induced bronchopneumonia injury in vitro and in vivo. Firstly, the viability and apoptosis of human bronchial epithelial BEAS-2B cells after luteolin treatment were assessed. Then, cells were treated with 10 mu M LPS to simulate inflammatory injury. The potential protective effects of luteolin on LPS-induced BEAS-2B cell inflammatory injury were detected. Moreover, after LPS and/or luteolin treatment, the expression of microRNA-132 (miR-132) in BEAS-2B cells was measured. The roles of miR-132 in protective activity of luteolin were investigated. Finally, the LPS-induced bronchopneumonia murine model was established and the anti-inflammatory effects of luteolin in vivo were analyzed. The results showed that LPS decreased BEAS-2B cell viability, increased cell apoptosis and enhanced inflammatory cytokines expression. Luteolin alleviated the LPS-induced viability loss, apoptosis and elevated expression of inflammatory cytokines in a dose-dependent manner. Moreover, luteolin alleviated the LPS-induced miR-132 expression increase in BEAS-2B cells. Overexpression of miR-132 reversed the protective effects of luteolin on LPS-induced inflammatory injury. Mechanistically, luteolin mitigated LPS-induced activation of NF-kappa B signaling pathway by down-regulation of miR-132. Furthermore, we also found that luteolin alleviated LPS-induced bronchopneumonia model in vivo. In conclusion, this study revealed that luteolin alleviated LPS-induced bronchopneumonia injury in vitro and in vivo through down-regulating miR-132. These findings provide theoretical basis for deeply exploring the treatment of bronchopneumonia in children by using luteolin.
The existence of drug resistance is the main reason for chemotherapeutic failure in malignancies. Long noncoding RNA (1ncRNA) maternally expressed gene 3 (MEG3) is implicated with tumorigenesis and chemoresistance. In the present study, we aimed to identify the role of MEG3 in oxaliplatin-resistant colorectal cancer (CRC) and its potential mechanisms. MEG3 was down-regulated in oxaliplatin-resistant CRC tissues and cell lines. Low MEG3 expression was correlated with poor prognosis of CRC patients. Overexpression of MEG3 improved oxaliplatin sensitivity of HT29/OXA and HCT116/OXA cells. MEG3 suppressed miR-141 expression in HCT116/OXA cells. Moreover, MEG3 elevated PDCD4 expression through targeting miR-141, acing as a competing endogenous RNA (ceRNA). miR-141 inhibition or PDCD4 up-regulation could mimic the functional role in oxaliplatin resistance, which was counteracted by overexpression of MEG3. Collectively, MEG3 facilitated the sensitivity of CRC cells to oxaliplatin by regulating miR-141/PDCD4 axis, providing a novel therapeutic strategy for CRC.
Long noncoding RNAs (LncRNAs) have been reported to play vital roles in non-small cell lung cancer (NSCLC). Recently, LncRNA/VPS9D1-AS1 has been reported to be overexpressed in various cancers. In this study, we aimed to investigate its expression pattern and clinical significance and further evaluate its prognostic value for NSCLC. VPS9D1-AS1 expression was examined in 184 NSCLC patients using a highly sensitive in situ hybridization protocol (RNAscope), and the expression values were correlated with the clinicopathological features. Another cohort including 12 NSCLC patients was used to validate the differential expression of VPS9D1-AS1 by qRT-PCR. TCGA datasets were further used to validate the main findings. We found that the levels of VPS9D1-AS1 were significantly higher in cancer tissues than in paired normal tissues from both lung squamous cell carcinoma (SCC) and adenocarcinoma (ADC) (P < 0.001). Importantly, the levels of VPS9D1-AS1 in patients with lung SCC were significantly higher than those in patients with lung ADC. The high levels of VPS9D1-AS1 were found to be associated with cancer lymph node metastasis (P = 0.020). Prognostic analysis revealed that the survival time for SCC patients with high levels of VPS9D1-AS1 was significantly shorter than that of patients with low levels of VPS9D1-AS1 (P = 0.007). Therefore, our findings suggest that the overexpression of VPS9D1-AS1 serves as a promising biomarker to predict the prognosis of NSCLC.
Aims : : Recent research showed that Long non-protein coding RNA ferritin heavy chain 1 pseudogene 3 (FTH1P3) plays a crucial role in the course of tumor formation. The present study was aimed to explore its role in esophageal squamous cell carcinoma (ESCC). Main methods: Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) was used to examine the expression levels of FTH1P3, mRNA SP1 and NF-kB in ESCC samples and cell lines. The impact of FTH1P3 knockdown was evaluated by WST-1 assays, colony formation assays, scratch wound assays, migration and invasion assays. Key findings: FTH1P3 was significantly upregulated in ESCC tissues and cells (P < 0.001). Knockdown of FTH1P3 notably decreased the proliferation, migration, and invasion capacity of ESCC cells. Silencing of FTH1P3 decreased the expression of specificity protein 1 (Sp1) and NF-kB (p65) in EC9706 and EC1. Significance: FTH1P3 plays a crucial role in ESCC tumorigenesis, and can be used as a potential therapeutic target for ESCC.
Araloside A is a triterpenoid saponin,which exhibits a broad spectrum of pharmacological activities, such as stimulating fibrinolysis, preventing coagulant, inhibiting renin, and decreasing blood pressure. Our previous report found that the compound exhibits a poor absolute bioavailability. However the underlying mechanisms of its absorption have not been investigated in the small intestine or in a Caco-2 cell model. In this study, the absorption mechanisms of araloside A were investigated in a Caco-2 cell monolayer and in a single-pass intestinal perfusion in situ model with Sprague-Dawley rats. The effects of basic parameters, such as compound concentration, time, temperature, paracellular pathway, different intestinal segments were analyzed, and the susceptibility of araloside A absorption process to treatment with various inhibitors, such as the P-gp inhibitor verapamil, the multidrug resistance protein2 inhibitors (MRP2) MK571 and indomethacin, the breast cancer resistance protein (BCRP) inhibitors Ko143 and reserpine, and endocytosis inhibitor chlorpromazine were assessed. It can be found that the mechanisms of intestinal absorption of araloside A may involve multiple transport pathways, such as passive diffusion, the paracellular pathway, as well as the participation of efflux transporters.
Burkitt's lymphoma is a type of highly aggressive Non-Hodgkin's lymphoma. Although advanced Burkitt's lymphoma is responsive to high-intensity chemotherapy regimens, increasing systemic toxicity, tumor recurrence and metastasis significantly reduce the patient survival. Thus, it is important to investigate novel antitumor agents with safety and effectiveness. beta-elemene shows anti-proliferative effect on cancer cells by triggering apoptosis through regulating several molecular signaling pathways. However, its role in the suppression of Burkitt's lymphoma has not yet been fully elucidated. The inhibitory effect of beta-elemene in Burkitt's lymphoma was studied in vitro and in vivo, as well as the involved molecular mechanism. The results demonstrated that beta-elemene effectively inhibited the growth and induced the apoptosis of Burkitt's lymphoma cells through upregulation of PUMA expression and modulating PUMA related apoptotic signaling pathway. The in vivo data confirmed the anti-tumor effect of beta-elemene in the xenografts, suggesting that beta-elemene is associated with PUMA activation, leading to Bax and caspase induction and onset of mitochondrial apoptosis.