Expression Of Lactate Dehydrogenase Research Articles

Overexpression of ascitic interleukin-35 induces CD8+ T cell exhaustion in liver cirrhotic patients with spontaneous bacterial peritonitis

Interleukin (IL) -35 induces immunotolerance by suppression of CD8+ T cells during chronic infections and cancers. In the present study, we amined to investigate the role of IL-35-mediated regulation of CD8+ T cells in patients with liver cirrhosis. Seventy-one patients with liver cirrhosis (46 patients with untainted ascites and 25 patients with spontaneous bacterial peritonitis [SBP]) and 22 controls were enrolled. Plasma and ascitic IL-35 levels were measured using ELISA. Peripheral and ascitic CD4+ and CD8+ T cells were purified to investigate their functional phenotypes. IL-35-stimulated CD8+ T cells were cultured with HepG2 cells in direct and indirect contact systems. Lactate dehydrogenase expression and cytokine secretion were measured to determine the cytotoxicity of CD8+ T cells. Plasma IL-35 was elevated in patients with liver cirrhosis, and ascitic IL-35 levels were higher in the SBP group than in the untainted ascites group. No significant differences in transcription factor expression or cytokine production in peripheral and ascitic CD4+ T cells were observed among groups. In the SBP group, ascitic CD8+ T cells expressed decreased cytotoxic molecules, along with the reduced secretion of interferon-γ and tumor necrosis factor-α when compared with the untainted ascites group. IL-35 stimulation suppressed ascitic CD8+ T cell cytotoxicity and cytokine production in both direct and indirect contact culture systems. This process was accompanied by decreased cytotoxic molecule expression and increased immune-checkpoint molecules in ascitic CD8+ T cells. The present findings revealed that overexpression of ascitic IL-35 dampened the cytotoxicity of CD8+ T cells in liver cirrhotic patients with SBP.

Effects of hypothyroidism on the female pancreas involve the regulation of estrogen receptors

This study aimed to investigate the impact of short-time hypothyroidism on the expression of aromatase, estrogen receptors (ERα, β), and GPR30 in the pancreas of female rabbits. The formation of new islets and the expression of insulin, GLUT4, and lactate dehydrogenase (LDH) were also analyzed. This purpose is based on actions that thyroid hormones and estrogens have on β-cells differentiation, acinar cell function, and insulin secretion. Twelve Chinchilla-breed adult virgin female rabbits were divided into control (n = 6) and hypothyroid (n = 6; methimazole 10 mg/kg for 30 days) groups. In the complete pancreas, expressions of aromatase and estrogen receptors, as well as proinsulin, GLUT4, and LDH were determined by western blot. Characteristics of islets were measured in slices of the pancreas with immunohistochemistry for insulin. Islet and acinar cells express aromatase, ERα, ERβ, and GPR30. Hypothyroidism increased the expression of ERα and diminished that for aromatase, ERβ, and GPR30 in the pancreas. It also promoted a high number of extra small islets (new islets) and increased the expression of proinsulin and GLUT4 in the pancreas. Our results show that actions of thyroid hormones and estrogens on β-cells neogenesis, acinar cell function, and synthesis and secretion of insulin are linked. Thus, the effects of hypothyroidism on the pancreas could include summatory actions of thyroid hormones plus estrogens. Our findings indicate the importance of monitoring estrogen levels and actions on the pancreas of hypothyroid women, particularly when serum estrogen concentrations are affected such as menopausal, pregnant, and those with contraceptive use.

Clinical Significance and Prognostic Value of Lactate Dehydrogenase Expression in Cervical Cancer.

Background: Lactate dehydrogenase (LDH) is a marker of injury and disease as it is expressed extensively in numerous cell types and tissues. Moreover it is released during tissue breakdown, and is elevated in cancerous tissues. However, the clinical significance and prognostic value of LDH as a tumor marker have been subject to considerable discussion. Objective: In this study, clinical serum LDH data from patients with cervical cancer (CC), CC microarray data, and RNA-seq data were integrated to assess the expression of LDH in CC. Methods: A total of 204 patients with newly diagnosed CC and 204 age-matched healthy controls were included to evaluate serum LDH levels in CC and non-cancer samples. External microarrays and RNA-seq datasets were collected for the differential expression analysis of LDH in CC and non-cancer tissue samples. Kaplan-Meier survival curves of the prognostic value of LDH for CC were plotted for RNA-seq data. Functional enrichment analysis was performed for the genes co-expressed with LDH. Results: The data from our in-house clinical cases as well as the data extracted from microarrays and RNA-seq databases demonstrated significant overexpression of LDH in CC samples. Elevated LDH expression levels were associated with poor overall survival in CC patients. The genes co-expressed with LDH were significantly correlated with the biological processes and pathways, associated with nuclear division, the condensed chromosome, protein serine/threonine kinase activity, and the cell cycle. Conclusion: In conclusion, LDH upregulation might serve as a therapeutic and prognostic biomarker for CC.

MRI-based radiomics models can improve prognosis prediction for nasopharyngeal carcinoma with neoadjuvant chemotherapy

BackgroundThe purpose of this study was to explore the prognostic value of imaging features and related models in nasopharyngeal carcinoma (NPC) patients that received neoadjuvant chemotherapy. Materials and methodsWe systematically reviewed the data of 110 NPC patients who received radiotherapy and neoadjuvant chemotherapy. The patients were randomly divided into the training cohort (n = 88) and the verification cohort (n = 22). The imaging data collected in this study were screened via Pyramidics and used to construct prediction models based on histology and clinical nomographs. The models' accuracy was evaluated via calibration curves and the consistency index (C-index). In addition, we also explored the correlation between radiomics expression patterns, quantitative histological characteristics, and clinical data and then constructed a model to predict the prognosis of NPC. ResultsThe models that integrated radiomics contours with all the clinical data were superior to those based on the clinical data alone (C-index 0.746 vs. C-index 0.814, respectively) and the calibration curves showed good consistency. The heat map showed that the radiomics expression pattern and selected histological characteristics were correlated with the clinical stage, T stage, and N stage (p < 0.05), and no radiomics feature was associated with lactate dehydrogenase expression, lymphocyte count, or mononuclear cell count. ConclusionMRI-based radiomics can significantly improve the efficacy of traditional TNM staging and clinical data in predicting the progression-free survival (PFS) of patients with advanced NPC, which may provide an opportunity for precision medicine.

Metabolic Phenotype of Daphnia Under Hypoxia: Macroevolution, Microevolution, and Phenotypic Plasticity

Daphnia is a freshwater crustacean that is able to upregulate hemoglobin (Hb) in response to hypoxia, imparting a red color. We combine multiple field surveys across season with a lab experiment to evaluate changes in the metabolic phenotype of Daphnia in relation to environmental hypoxia. Looking at the zooplankton community, we found that D. pulicaria was restricted to lakes with a hypoxic hypolimnion. Comparing D. pulicaria with different amounts of Hb, red animals showed higher mRNA levels for several Hb genes, whereas most glycolytic genes showed red/pale differences of less than 50%. We also observed seasonal changes in the metabolic phenotype that differed between red and pale animals. Hb was upregulated early in the season in hypoxic lakes, and a relationship between Hb and lactate dehydrogenase only emerged later in the season in a temporal pattern that was lake specific. To evaluate whether these differences were due to specific lake environments or microevolutionary differences, we tested the induction of genes under controlled hypoxia in isofemale lines from each of four lakes. We found a strong response to 18 h hypoxia exposure in both Hb and lactate dehydrogenase mRNA, although the magnitude of the acute response was greater than the steady state differences in mRNA levels between pale and red Daphnia. The baseline expression of Hb and lactate dehydrogenase also varied between isofemale lines with different lake origins. These results, in combination with comparison of glycogen measurements, suggests that Hb functions primarily to facilitate oxygen delivery, mitigating systemic hypoxia, rather than an oxygen store. The combination of lab and field studies suggest that the metabolic phenotype of the animal is influenced by both microevolutionary differences (within and between lakes) as well as the spatial and temporal environmental heterogeneity of the lakes. The differences between Daphnia species, and the unexpected lack of hypoxia sensitivity of select glycolytic genes provide evidence of macroevolutionary differences in metabolic strategies to cope with hypoxia.

Metformin alleviates the depression-like behaviors of elderly apoE4 mice via improving glucose metabolism and mitochondrial biogenesis

Apolipoprotein E4 (apoE4) is closely related to late-onset depression (LOD). In addition, the benefits of metformin treatment of depression have been documented in a range of rodent studies and human trials, but few studies have probed into the effect of metformin on and the related mechanism in depressed elderly mice, especially in those APOE4 carriers. Here, we treated 13-month-old apoE3-targeted replacement (TR) and apoE4-TR mice with an intragastric administration of metformin (300 mg/kg/d) or normal saline for 5 months. We found that metformin exerted antidepressant effects on apoE4 mice, including reduced immobility time in TST and FST, and increased ratios of time and distance in the central area of OFT. Importantly, compared with apoE3 mice, apoE4 mice showed a higher expression of lactate dehydrogenase (LDH) and pyruvate dehydrogenase kinase (PDK1 and PDK4) in the hippocampus. The increased LDH level was rescued by metformin treatment. Moreover, the metformin administration increased the levels of transcriptional factor NRF-1 and TFAM, mtDNA, and most mitochondrial complex subunits in apoE-TR mice. Furthermore, it upregulated the expressions of antioxidant enzymes, such as MnSOD, GPX1, and GSR1/2. Interestingly, apoE4 blunted the hypoglycemic effect of metformin in aged mice. These data suggest that metformin ameliorates the depression-like behaviors probably by improving glucose metabolism and mitochondria biogenesis in the hippocampus of aged apoE4 mice. These findings imply that chronic metformin treatment can improve apoE4-mediated LOD, providing mechanistic insights for apoE4- and age-based depression prevention and therapy.

Using gene expression to identify the most suitable environmental conditions for growth and metabolism of juvenile deepwater redfish (Sebastes mentella) in the Estuary and the Gulf of St. Lawrence

Abstract Deepwater redfish Sebastes mentella will be among the most important resource-sustaining commercial bottom-fish fisheries in the years to come in the Estuary and Gulf of St. Lawrence (EGSL). In 2011, 2012, and 2013, three strong cohorts were recruited to the stock; their abundance in 2018 was 80 times higher than that of the 1993–2012 period. The main goal of this work was to deepen our knowledge of their growth regulation and metabolism in order to identify molecular indicators and determine how they are influenced by natural environmental conditions. Fish weight and water temperature explained 11% of the variation in relative mRNA levels of specific gene targets in liver and muscle among seven sites where deepwater redfish were captured in the EGSL. The relative expression of liver insulin-like growth factor-1 (igf-1) and white muscle A-chain lactate dehydrogenase (ldh-a) correlate positively with weight, whereas heavy chain muscle myosin (myo), heart citrate synthase (cs), and white muscle pyruvate kinase (pk) correlate negatively. The relative expression of heart cytochrome c oxidase subunit 1 (cox-1) and white muscle igf-1 receptor isoform a (igf-1ra) correlate negatively with temperature. Deepwater redfish from the estuary were smaller than those caught at other sites. Since the growth potential of deepwater redfish was strongly correlated with temperature (being enhanced by higher temperatures), this study suggests an ecological advantage for this species in a climate-warming context.

Expression of Receptor Activator of Nuclear – Kappa β Ligand in Patients with Metastatic Bone Disease

BACKGROUND: Bone metastasis is a complication that often occurs due to cancer in solid organs, and more often compared to primary bone tumors. Bone metastasis is associated with excessive osteolytic processes. AIM: This study was conducted to reveal the correlation between Receptor Activator of Nuclear factor kβ Ligand (RANKL) expression, lactate dehydrogenase (LDH), and neutrophil-lymphocyte ratio (NLR) level in patients with bone metastatic lesions. METHODS: This cross-sectional study was conducted in 15 subjects with bone metastatic lesions. The specimens were fresh bone tissues obtained by open biopsy. Expression of RANKL in mRNA level was detected quantitatively using reverse transcription-polymerase chain reaction. LDH and NLR were analyzed from the peripheral blood analysis. The correlation of RANKL expression with LDH and NLR was statistically analyzed. RESULTS: This study enrolled 15 subjects with bone metastasis disease based on the clinical, radiological, and histopathological results. The means of LDH, NLR, and RANKL expression were not significantly different. There was no significant association between the characteristics of metastatic lesion and RANKL expression. The correlation between LDH levels with RANKL expression was positive (p &lt; 0.05), while the correlation between NLR and RANKL expression was negative (p &lt; 0.05). CONCLUSION: RANKL expression was positively correlated with LDH and negatively correlated with the NLR. Both LDH and NLR can be used as predictive factors of RANKL expression in bone metastasis.

Molecular Investigations of the Effect of Thermal Manipulation During Embryogenesis on the Enzymatic Activity of Creatine Kinase and Lactate Dehydrogenase in Broiler Muscle

Incubating temperature and timing or duration is critical to determine the optimum protocol of thermal manipulation (TM), which underlines muscle growth improvement. Therefore, the aim of the present study is to determine the optimum period of embryonic TM that may result in the improvement of pectoral and thigh muscle myogenesis. This is done by investigating the level of mRNA expression of creatine kinase (CK) and lactate dehydrogenase (LDH). An additional goal is measuring the blood levels of CK and LDH as a biomarker of muscle injury due to the experimental thermal challenge on post-hatch day 35. The study was conducted on 1,440 fertile eggs (Ross broilers) that were divided randomly and equally into a control group and four treatment groups (TM1, TM2, TM3, and TM4). The treatment groups were daily subjected to TM at 39 ºC for 18h with 65% relative humidity (RH) during embryonic days (EDs) 7-11, 11-15, 15-18, and 7-18, respectively. Among the thermally manipulated groups that were investigated, TM1 (ED 7-11) resulted in significant improvement of mRNA expression and enzymatic concentration of CK and LDH in muscle during embryogenesis, as compared to the control. Six hours of TC showed the highest significant CK and LDH expression and concentration levels in the control as compared to TM groups. Thus, the results of this study indicate that TM during ED 7-11 improves pectoral and thigh muscles’ response to heat stress without adversely affecting their performance. This finding could be used by commercial breeders to enhance local broiler production.

MicroRNA-124a Protects the Myocardium Against Ischemia Reperfusion Injury Through Regulation of the Notch Signaling Pathway.

IntroductionThis study’s objective is to investigate the effect of downregulation of micro ribonucleic acid (miR)-124a on myocardial injury after ischemia reperfusion (I/R) in rats.MethodsSprague Dawley (SD) rats (n=20) were divided into four groups - sham, I/R, I/R+miR-124a antagomir (I/R+ant-miR-124a), and I/R+ant-normal control (NC). The pathomorphological and infarct size variance of injured myocardial tissues with IR were conducted with hematoxylin (HE) and triphenyltetrazolium chloride (TTC) staining. The expression levels of miR-124a, BAX, nuclear factor kappa B (NF-KB), Notch1, and Hes1 were examined by quantitative real-time polymerase chain reaction or Western blot in myocardium. The inflammatory cytokines interleukin (IL)-6, IL-1β, and tumor necrosis factor alpha (TNF-α) were detected by the enzyme-linked immunosorbent assay, as well as the activity of lactate dehydrogenase (LDH) and creatine kinase (CK) in serum by colorimetry.ResultsThe expression of miR-124a was increased in the I/R group. Compared with I/R and I/R+ant-NC groups, after downregulating miR-124a, the expression of IL-6, IL-1β, TNF-α, BAX, NF-KB, LDH, and CK were decreased, but the expression of Notch1 and Hes1 were increased. In HE staining, myocardial tissue edema, red blood cell exudation, and myocardial fiber arrangement disorder were accompanied by inflammatory cell infiltration and local necrosis in the I/R group. However, the pathological injury of myocardial tissue was alleviated after downregulating miR-124a. Additionally, TTC results showed that the myocardial infarction area was decreased in the I/R+ant-miR-124a group.ConclusionDownregulation of miR-124a expression through Notch pathway can significantly reduce myocardial damage after 24 hours of I/R in SD rats. Therefore, miR-124a may become a potential therapeutic target for I/R injury.

Effects of Panax notoginseng Saponin on Neurons Based on Oxygen-Glucose Deprivation/Reoxygenation after Focal Cerebral Ischemia

We aimed to evaluate the effects of Panax notoginseng saponin on neuronal apoptosis and neuroprotection based on oxygen-glucose deprivation/reoxygenation in rats with focal cerebral ischemia. 100 rats were randomly divided into sham operation, middle cerebral artery occlusion model, low-dose Panax notoginseng saponin (10 mg/kg), medium-dose Panax notoginseng saponin (25 mg/kg) and high-dose Panax notoginseng saponin (50 mg/kg) groups. The model of focal cerebral ischemia was established by middle cerebral artery occlusion. For sham group, the right common, external and internal carotid arteries were only dissociated, without ligation or insertion. After modeling, Panax notoginseng saponin groups were intraperitoneally injected with Panax notoginseng saponin. Hippocampal neurons isolated from normal rats were randomly divided into control, oxygen-glucose deprivation/reoxygenation model, 1 μmol/l, 5 μmol/l and 20 μmol/l oxygen-glucose deprivation/reoxygenation+Panax notoginseng saponin groups. Neurological function was scored by Longa method and neuron morphology was observed. Neuronuclear antigen and neuroepithelial stem protein in brain tissues were detected by immunohistochemistry. The survival rate of neurons, lactate dehydrogenase leakage, neuronal apoptosis and expression levels of protein kinase B, phosphorylatedprotein kinase B and apoptosis-related proteins B-cell lymphoma 2, Bcl-2-associated X protein and caspase-3 were examined by cell counting kit-8 assay, lactate dehydrogenase kit, Hoechst 33 342 staining and Western blotting, respectively. Compared with model group, Panax notoginseng saponin groups had significantly lower neurological function scores (p<0.05). Panax notoginseng saponin significantly relieved neuronal injury and increased neuronuclear and neuroepithelial stem protein-positive cells (p<0.05). It significantly raised the survival rate of neurons, reduced lactate dehydrogenase leakage and inhibited neuronal apoptosis dose-dependently. Panax notoginseng saponin up-regulated the protein expressions of phosphorylated-protein kinase B/protein kinase B and B-cell lymphoma 2 and down-regulated those of Bcl-2-associated X protein and caspase-3 dose-dependently. It can protect against and repair neuronal injury caused by oxygen-glucose deprivation/reoxygenation, probably by activating the phosphatidylinositol 3-kinase/protein kinase B signaling pathway to suppress neuronal apoptosis.

Co-delivery of photosensitizer and diclofenac through sequentially responsive bilirubin nanocarriers for combating hypoxic tumors

Considering that photodynamic therapy (PDT)-induced oxygen consumption and microvascular damage could exacerbate hypoxia to drive more glycolysis and angiogenesis, a novel approach to potentiate PDT and overcome the resistances of hypoxia is avidly needed. Herein, morpholine-modified PEGylated bilirubin was proposed to co-deliver chlorin e6, a photosensitizer, and diclofenac (Dc). In acidic milieu, the presence of morpholine could enable the nanocarriers to selectively accumulate in tumor cells, while PDT-generated reactive oxidative species (ROS) resulted in the collapse of bilirubin nanoparticles and rapid release of Dc. Combining with Dc showed a higher rate of apoptosis over PDT alone and simultaneously triggered a domino effect, including blocking the activity and expression of lactate dehydrogenase A (LDHA), interfering with lactate secretion, suppressing the activation of various angiogenic factors and thus obviating hypoxia-induced resistance-glycolysis and angiogenesis. In addition, inhibition of hypoxia-inducible factor-1α (HIF-1α) by Dc alleviated hypoxia-induced resistance. This study offered a sequentially responsive platform to achieve sufficient tumor enrichment, on-demand drug release and superior anti-tumor outcomes in vitro and in vivo.

Panchvalkal (polyherbal formulation) mitigates STZ induced type 2 DM by modulating the expression of hexokinase (HX), lactate dehydrogenase (LDH), triphosphate isomerase (TPI)

BackgroundThe present study was designed to evaluate the potentiality of Panchvalkal (Ayurvedic formulation) for the treatment of type 2 Diabetes Mellitus. Panchvalkal is polyherbal formulation of bark Ficus benghalensis L., Ficus racemosa L., Ficus religiosa L., Thespesia populnea L. and Ficus lacor Buch-Ham. The formulation was traditionally used as anti-inflammatory, antibacterial and to treat ulcers, wounds, leucorrhoea and other vaginal diseases. ObjectiveThe current study was hypothesized to investigate Panchvalkal aqueous extract protective property against streptozotocin induced Diabetes Mellitus Swiss albino rats and its effects on HX, LDH and TPI genes involved in glycolysis pathway. MethodAqueous extract of the bark of five medicinal plants were prepared. The aqueous extract of Panchvalkal was analyzed for antioxidant property (TF, TP, DPPH and RP). Diabetes was induced in Swiss albino rats through streptozotocin (60 mgKg-1 bw i.p.) administration. The extract (500 mgKg-1 bw) and glibenclamide (300 mgKg-1 bw) alone and in combination was administrated orally to specific group of animals up to 28 days. Animals were scarified, blood was collected, and liver excised. cDNA was prepared from RNA isolated from the liver. Expression of HX, LDH and TPI were studied through real-time PCR analysis. ResultPanchvalkal aqueous extract was enriched with phenol content (82.84 mg GAE G−1) and flavonoid content (283.52 mg RE G−1) and showed effective antioxidant property. No sign of toxicity and mortality in rats were recorded during toxicity assessment. Rats administrated with Panchvalkal 500 mgKg-1 bw showed significant reduction in blood glucose, total cholesterol, triglycerides, LDL and increase in HDL level. Also the plasma glucose level was significantly (p < 0.05) reduced in comparison to diabetic control. Significant increase in enzymes level such as SGOT and SGPT was observed. Panchvalkal modulated the expression level of Hexokinase (HX), Lactate dehydrogenase (LDH) and Triphosphate isomerase (TPI) genes in STZ induced T2DM. ConclusionThe present study showed that Panchvalkal acts effectively in modulating the mRNA expression of HX, LDH and TPI genes involved in the glycolysis pathway and other metabolic reactions. It significantly increased the glucose uptake and improved the glucose metabolism in T2DM.

Regulation of CcpA on the growth and organic acid production characteristics of ruminal Streptococcus bovis at different pH

BackgroundCatabolite control protein A (CcpA) regulates the transcription of lactate dehydrogenase and pyruvate formate-lyase in Streptococcus bovis, but knowledge of its role in response to different pH is still limited. In this study, a ccpA-knockout strain of S. bovis S1 was constructed and then used to examine the effects of ccpA gene deletion on the growth and fermentation characteristics of S. bovis S1 at pH 5.5 or 6.5.ResultsThere was a significant interaction between strain and pH for the maximum specific growth rate (μmax) and growth lag period (λ), which caused a lowest μmax and a longest λ in ccpA-knockout strain at pH 5.5. Deletion of ccpA decreased the concentration and molar percentage of lactic acid, while increased those of formic acid. Strains at pH 5.5 had decreased concentrations of lactic acid and formic acid compared to pH 6.5. The significant interaction between strain and pH caused the highest production of total organic acids and acetic acid in ccpA-knockout strain at pH 6.5. The activities of α-amylase and lactate dehydrogenase decreased in ccpA-knockout strain compared to the wild-type strain, and increased at pH 5.5 compared to pH 6.5. There was a significant interaction between strain and pH for the activity of acetate kinase, which was the highest in the ccpA-knockout strain at pH 6.5. The expression of pyruvate formate-lyase and acetate kinase was higher in the ccpA-knockout strain compared to wild-type strain. The lower pH improved the relative expression of pyruvate formate-lyase, while had no effect on the relative expression of acetate kinase. The strain × pH interaction was significant for the relative expression of lactate dehydrogenase and α-amylase, both of which were highest in the wild-type strain at pH 5.5 and lowest in the ccpA-knockout strain at pH 6.5.ConclusionsOverall, low pH inhibited the growth of S. bovis S1, but did not affect the fermentation pattern. CcpA regulated S. bovis S1 growth and organic acid fermentation pattern. Moreover, there seemed to be an interaction effect between pH and ccpA deletion on regulating the growth and organic acids production of S. bovis S1.

LncRNA MIAT promotes hypoxia-induced H9C2 cell pyroptosis via binding to SF1 to inhibit CGRP transcription.

What is the central question of this study? How does long non-coding RNA myocardial infarction-associated transcript (lncRNA MIAT) function in hypoxia-induced H9C2 cells? What is the main finding and its importance? LncRNA MIAT inhibited transcription of calcitonin gene-related peptide by binding to splicing factor1, thereby promoting hypoxia-induced H9C2 cell pyroptosis. This study provides a new theoretical basis for the treatment of acute myocardial infarction by using lncRNA MIAT as a molecular target to mediate cardiomyocyte pyrodeath. Hypoxia induces severe cardiomyocyte pyroptosis, contributing to acute myocardial infarction. The aim of this study was to analyse the molecular mechanism of long non-coding RNA myocardial infarction-associated transcript (lncRNA MIAT) in hypoxia-induced H9C2 cell pyroptosis. A hypoxic H9C2 cell model was established. Cell viability was detected via the Cell Counting Kit-8 method. Levels of lactate dehydrogenase, malondialdehyde and superoxide dismutase and expressions of pyroptotic markers, lncRNA MIAT, splicing factor1 (SF1) and calcitonin gene-related peptide (CGRP) were detected via qRT-PCR. The subcellular localization of lncRNA MIAT was predicted and confirmed via LncATLAS and nuclear/cytosol fractionation assay. The binding relationships between lncRNA MIAT and SF1 and between SF1 and the CGRP promotor were verified via RNA immunoprecipitation. Rescue experiments were designed to confirm the role of lncRNA MIAT/SF1/CGRP in H9C2 cell pyroptosis. LncRNA MIAT was overexpressed in hypoxia-induced H9C2 cells. Hypoxia induced pyroptosis in H9C2 cells. Silencing of lncRNA MIAT enhanced cell viability and alleviated pyroptosis. LncRNA MIAT inhibited CGRP transcription via binding to SF1. Overexpression of SF1 promoted CGRP transcription and relieved H9C2 cell pyroptosis. Downregulation of CGRP reversed the role of silencing lncRNA MIAT in H9C2 cell pyroptosis. Overall, lncRNA MIAT inhibited CGRP transcription via binding to SF1, thereby promoting hypoxia-induced H9C2 cell pyroptosis.

Starvation induced autophagy promotes the progression of bladder cancer by LDHA mediated metabolic reprogramming

BackgroundAberrant autophagy and preternatural elevated glycolysis are prevalent in bladder cancer (BLCA) and are both related to malignant progression. However, the regulatory relationship between autophagy and glycolytic metabolism remains largely unknown. We imitated starvation conditions in the tumour microenvironment and found significantly increased levels of autophagy and aerobic glycolysis, which both regulated the progression of BLCA cells. We further explored the regulatory relationships and mechanisms between them.MethodsWe used immunoblotting, immunofluorescence and transmission electron microscopy to detect autophagy levels in BLCA cells under different treatments. Lactate and glucose concentration detection demonstrated changes in glycolysis. The expression of lactate dehydrogenase A (LDHA) was detected at the transcriptional and translational levels and was also silenced by small interfering RNA, and the effects on malignant progression were further tested. The underlying mechanisms of signalling pathways were evaluated by western blot, immunofluorescence and immunoprecipitation assays.ResultsStarvation induced autophagy, regulated glycolysis by upregulating the expression of LDHA and caused progressive changes in BLCA cells. Mechanistically, after starvation, the ubiquitination modification of Axin1 increased, and Axin1 combined with P62 was further degraded by the autophagy–lysosome pathway. Liberated β-catenin nuclear translocation increased, binding with LEF1/TCF4 and promoting LDHA transcriptional expression. Additionally, high expression of LDHA was observed in cancer tissues and was positively related to progression.ConclusionOur study demonstrated that starvation-induced autophagy modulates glucose metabolic reprogramming by enhancing Axin1 degradation and β-catenin nuclear translocation in BLCA, which promotes the transcriptional expression of LDHA and further malignant progression.

Opicapone Protects Against Hyperhomocysteinemia-Induced Increase in Blood-Brain Barrier Permeability.

Hyperhomocysteinemia (HHcy)-related brain vascular disorders and brain endothelial dysfunction are important characteristics of the pathogeneses of subarachnoid hemorrhage and stroke. Upregulated homocysteine (Hcy) can impair the integrity of the blood-brain barrier (BBB). Opicapone has been recently licensed for the management of Parkinson's disease (PD); however, it is unknown whether it possesses a protective effect in brain vessels against HHcy. To investigate the beneficial effects of Opicapone on BBB permeability against HHcy, we carried out both in vivo and in vitro experiments. Mice were allocated into four groups: the Control, Opicapone, HHcy, and HHcy + Opicapone. Interestingly, we found that the administration of Opicapone attenuated the increased BBB permeability in Hcy-treated mice, as determined by sodium fluorescein staining. The immunofluorescence staining showed that Opicapone prevented homocysteine-induced reduction of claudin-2 in the mice cortices. The in situ zymography assay revealed that Opicapone suppressed homocysteine-increased matrix metalloproteinases (MMPs) activity in the cortices. In bEnd.3 brain endothelial cells, Opicapone treatment ameliorated homocysteine-induced lactate dehydrogenase (LDH) release and expression of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9). Furthermore, Opicapone alleviated homocysteine-induced decrease in claudin-2 level in bEnd.3 cells. In summary, our results show that Opicapone protects against HHcy-induced BBB permeability by reducing the expression and gelatinase activity of MMPs, and increasing the expression of claudin-2.

P53-Independent Effects of Set7/9 Lysine Methyltransferase on Metabolism of Non-Small Cell Lung Cancer Cells.

Set7/9 is a lysine-specific methyltransferase, which regulates the functioning of both the histone and non-histone substrates, thereby significantly affecting the global gene expression landscape. Using microarray expression profiling, we have identified several key master regulators of metabolic networks, including c-Myc, that were affected by Set7/9 status. Consistent with this observation, c-Myc transcriptional targets—genes encoding the glycolytic enzymes hexokinase (HK2), aldolase (ALDOB), and lactate dehydrogenase (LDHA)—were upregulated upon Set7/9 knockdown (Set7/9KD). Importantly, we showed the short hairpin RNA (shRNA)-mediated attenuation of Set7/9 augmented c-Myc, GLUT1, HK2, ALDOA, and LDHA expression in non-small cell lung cancer (NSCLC) cell lines, not only at the transcriptional but also at the protein level. In line with this observation, Set7/9KD significantly augmented the membrane mitochondrial potential (MMP), glycolysis, respiration, and the proliferation rate of NSCLC cells. Importantly, all these effects of Set7/9 on cell metabolism were p53-independent. Bioinformatic analysis has shown a synergistic impact of Set7/9 together with either GLUT1, HIF1A, HK2, or LDHA on the survival of lung cancer patients. Based on these evidence, we hypothesize that Set7/9 can be an important regulator of energy metabolism in NSCLC.

Experimental Research on Intratumoral Injection of Sal Ammoniac Extract in the Treatment of Mouse Liver Cancer

Objective: To study the effect of sal ammoniac extract on the treatment of liver cancer and analyze its possibility of replacing absolute ethanol. Methods: Sixty Kunming mice (5-6 weeks old, 18-22g in weight, male and female in half) were selected and inoculated with 0.1 ml of 1:4 ascitic diluent from mouse liver cancer H22 under the axilla of the right limb. After tumor formation, they were randomly divided into 3 groups with 20 mice in each group. Normal saline (NS), sal ammoniac extract (N), and absolute ethanol (E) were injected into the tumor once a day for four times. The death, tumor weight, tumor inhibition rate, lactate dehydrogenase (LDH), and succinate dehydrogenase (SDH) expression of the mice in each group were analyzed. Results: In the course of treatment, 3 mice died in the normal saline group, 2 mice died in the sal ammoniac treatment group, and 7 mice died in the absolute ethanol group. The tumor weight of the normal saline group, sal ammoniac treatment group, and absolute ethanol group were 1.88 ± 0.26, 1.41 ± 0.49, and 1.51 ± 0.46, respectively. The tumor inhibition rates were 0%, 31.0%, and 21.6%, respectively. Comparing the sal ammoniac treatment group and the normal saline group, t = 3.5133, p = 0.0013; comparing the absolute ethanol group and the normal saline group, t = 2.7926, p = 0.0093. The expression of LDH was 81 ± 10, 51 ± 11, and 32 ± 9 in the normal saline group, sal ammoniac treatment group, and absolute ethanol group, respectively, while the expression of SDH was 80 ± 10, 51 ± 10, and 51 ± 12 in the normal saline group, sal ammoniac treatment group, and absolute ethanol group, respectively. Comparing the LDH of the sal ammoniac treatment group with that of the saline group, t = 8.4264, p = 0.0000; comparing the absolute ethanol group and normal saline group, t = 13.8763, p = 0.0000. Comparing the SDH of the sal ammoniac treatment group with that of the normal saline group, t = 8.1455, p = 0.0000; comparing the absolute ethanol group with the normal saline group, t = 7.2197, p = 0.0000. Conclusion: The traditional Chinese antitumor medicine, sal ammoniac and its main effective components have good antitumor effect, which can be further popularized and applied in clinical practice.

Targeting the tumor metabolism by oxamate potentiates the impact of chemotherapeutics in colorectal cancer cells

Background/Aim: Cancer cells promote lactate formation via pyruvate rather than oxidative phosphorylation by programming their metabolism to maintain proliferation under the Warburg effect. It has shown that the altered metabolic phenotype with activation of lactate dehydrogenase-A in the cancer cell may affect survival, chemotherapy resistance, and metastasis. In this direction, studies are focusing on reprogramming cancer metabolism and increase the effectiveness of chemotherapy. In this study, the main aim was to target the Warburg phenotype via the inhibition of lactate dehydrogenase with the combination of sodium oxamate and current colorectal cancer treatment options such as 5-fluorouracil and irinotecan. &#x0D; Methods: The effect of chemotherapeutics on the cellular behaviour was evaluated by real-time cytotoxicity and migration analysis systems, and metabolic phenotype was assessed by measuring lactate, lactate dehydrogenase expression, and reactive oxygen species levels. &#x0D; Results: According to the results, the viability and migration of colorectal cancer cells were significantly decreased with the combination of chemotherapeutics and sodium oxamate which decreases lactate levels. &#x0D; Conclusion: As a result, the combination of sodium oxamate with the chemotherapeutics hinders the cancer cell viability and migration by changing metabolic phenotype with decreased lactate.