ABSTRACT
The most common type of intracranial malignancy is glioma. Although the current treatments are surgery, radiation and chemotherapy, the prognosis for patients with glioma is not promising. Therefore, it becomes critical to find an effective management. The literature shows that microRNA (miRNA) plays an important role in tumorigenesis and progression. Based on this, this study aimed to investigate the molecular mechanism of miR-525-5p in regulating the migration, invasion and proliferation of glioma cells. The TCGA database was used to identify perilipin 3 (PLIN3) differentially expressed in normal tissues and glioma tissues, and the CGGA and GEPIA databases were used to query that high expression of PLIN3 was associated with poor prognosis in glioma patients and Western blot experiments revealed that PLIN3 was highly expressed in glioma cells (P<0. 05) . The results of wound healing assay and Transwell invasion assay showed that knockdown or overexpression of PLIN3 respectively inhibited or promoted the migration and invasion of glioma cells (P < 0. 05) . Dual luciferase assays confirmed that PLIN3 could bind to miR-525-5p target. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) showed that miR-525-5p expression was lower in LN229 and U251 glioma cells than in human astrocyte (HA) (P < 0. 05) . Transwell assay and 5-ethynyl-2-deoxyuridine (EdU) cell proliferation assay verified that down- or up-regulation of miR-525-5p could reverse the effects of overexpression or knockdown of PLIN3 on LN229 glioma cells (P<0. 05) . Taken together, miR-525-5p was able to regulate the migration, invasion and proliferation of glioma cells by targeting PLIN3.
ABSTRACT
SUMMARY OBJECTIVE: Heart attack is one of the most common causes of sudden death in adults. Therefore, early detection of heart attack and investigation of potential new biomarkers are of great importance. We investigated whether perilipin-5 is a potential biomarker by examining changes in perilipin-5 serum levels along with high-sensitivity cardiac troponin I during a heart attack. METHODS: The subjects were divided into two groups: (1) control group and (2) patients with heart attack, with 150 people in each group. High-sensitivity cardiac troponin I, perilipin-5, total oxidant status, malondialdehyde, reduced glutathione, and superoxide dismutase levels in serum samples were measured. In addition, perilipin-5 mRNA expressions and protein levels were analyzed. RESULTS: There was no overall statistical difference between the demographic characteristics of the groups. However, high-density lipoprotein, creatine kinase, Creatine kinase myocardial band, aspartate amino transferase, lactate dehydrogenase, and calcium levels were higher in the heart attack group compared to the control group. We found that the high-sensitivity cardiac troponin I and perilipin-5 levels increased in the patients with heart attack (p<0.0001) compared to control. Although there was an insignificant increase in malondialdehyde levels in the heart attack group (p>0.05), there was a 35.9% increase in total oxidant status levels and a 33.5 and 24.1% decrease in glutathione and superoxide dismutase levels, respectively (p<0.01), compared to control. Perilipin-5 mRNA and protein levels in heart attack patients increased by 48.2 and 23.6%, respectively, compared to the control group (p<0.01). CONCLUSION: Our results showed that perilipin-5 together with high-sensitivity cardiac troponin I could be a promising biomarker in heart attack.
ABSTRACT
Objective:To analyze the correlation between serum ferritin (SF), perilipin, leptin and the outcome of patients with gestational diabetes mellitus (GDM).Methods:From October 2017 to December 2019, 126 patients with GDM who underwent maternity checkups in Baoding Fourth Central Hospital and gave birth were selected as the GDM group, and 82 normal pregnant women during the same period were selected as the control group for retrospective analysis. The levels of serum SF, perilipin, and leptin in the GDM group and the control group were measured and compared, and the expressions of serum indexes of patients with different blood glucose control and different pregnancy outcomes in the GDM group were measured. Pearson correlation analysis was used to explore the correlation between the expression of serum indexes in GDM patients and fasting blood glucose (FPG), 2 h postprandial blood glucose (2 h PG), and insulin resistance index (HOMA-IR). Logistic regression was used to analyze the relationship between the expression of serum indicators and adverse pregnancy outcomes in GDM patients. The receiver operating characteristic (ROC) curve was used to observe the value of single serum indexes and to predict the pregnancy outcome of GDM patients.Results:The levels of SF, perilipin and leptin in GDM group were higher than those in control group: (152.48 ± 37.64) μg/L vs. (109.27 ± 32.16) μg/L, (857.06 ± 192.35) ng/L vs. (262.83 ± 104.7) ng/L, (23.54 ± 2.28) μg/L vs.(14.62 ± 1.83) μg/L, the differences were statistically significant ( P<0.05). The levels of SF, perilipin and leptin in patients with good blood glucose control in GDM group were lower than those in patients with poor blood glucose control: (132.10 ± 36.52) μg/L vs. (176.37 ± 40.06) μg/L, (176.37 ± 40.06) ng/L vs. (946.42 ± 205.37) ng/L, (21.49 ± 2.16) μg/L vs. (25.94 ± 2.40) μg/L, the differences were statistically significant ( P<0.05). The levels of serum SF, perilipin and leptin in GDM patients were positively correlated with FPG, 2 h PG and HOMA-IR levels ( P<0.05). The levels of serum SF, perilipin and leptin in GDM patients with adverse pregnancy outcomes were higher than those in patients without occurrence: (182.86 ± 42.29) μg/L vs. (138.86 ± 35.47) μg/L, (1 013.35 ± 216.07) ng/L vs. (787.00 ± 183.49) ng/L, (27.04 ± 2.5) μg/L vs. (21.97 ± 2.07) μg/L, the differences were statistically significant ( P<0.05). Logistic regression analysis showed that serum SF, perilipin and leptin were closely related to the adverse pregnancy outcomes in GDM patients ( P<0.05). ROC curve analysis showed that the sensitivity and the specificity of SF, perilipin, leptin combined detection to predict GDM patients with adverse pregnancy outcomes was 76.92% and 83.91%. Conclusions:The serum SF, perilipin and leptin are abnormally high expression in GDM patients, and it is positively correlated with blood glucose level and HOMA-IR. Joint detection can improve the predictive value of adverse pregnancy outcomes and provide a basis for early intervention.
ABSTRACT
Differentiation of preadipocyte, also named adipogenesis, leads to the phenotype of mature adipocyte that is filled with many lipid droplets. Excessive lipid accumulation in adipocytes leads to the development of obesity. In this study, we investigated the effect of 11 different natural compounds on lipid accumulation during the differentiation of 3T3-L1 preadipocytes into 3T3-L1 adipocytes. Strikingly, among the natural compounds, cryptotanshinone at 10 µM most strongly reduced triglyceride (TG) contents in 3T3-L1 cells after 8 days of the differentiation. Furthermore, cryptotanshinone at 10 µM significantly suppressed lipid accumulation in 3T3-L1 cells after 8 days of the differentiation. Cryptotanshinone at 1 to 10 µM tested did not affect the survival of 3T3-L1 cells after 8 days of the differentiation. On mechanistic levels, cryptotanshinone time-differentially decreased the expression levels of CCAAT/enhancer-binding protein-α (C/EBP-α), peroxisome proliferator-activated receptor-γ (PPAR-γ), fatty acid synthase (FAS), and perilipin A but also the phosphorylation levels of signal transducer and activator of transcription-3 (STAT-3) during the 3T3-L1 cell differentiation. Taken together, these findings demonstrate that cryptotanshinone inhibits lipid accumulation in differentiating 3T3-L1 cells, which appears to be mediated through the reduced expression and/or phosphorylation levels of C/EBP-α, PPAR-γ, FAS, Perilipin A, and STAT-3.
Subject(s)
3T3-L1 Cells , Adipocytes , Adipogenesis , Lipid Droplets , Obesity , Peroxisomes , Phenotype , Phosphorylation , Transducers , TriglyceridesABSTRACT
Objective To investigate the effects and mechanisms of perilipin-5 (Plin5) on the apoptosis of mouse cardiac microvascular endothelial cells induced by high fat and high glucose.Methods The mouse cardiac microvascular endothelial cells (MCMECs) cultured with high glucose medium were respectively given 0,100,300 and 500μmol/L palmitic acid for 24 hours.In order to explore the effects and mechanisms of Plin5 on MCMECs injuries induced by high fat and high glucose,MCMECs exposed to 300μmol/L palmitic acid for 24 hours were divided into control group,Scra siRNA group and Plin5 siRNA group.The control group was only treated with transfection reagent,the Scra siRNA group was given treatment of transfection reagent and garbled RNA,the Plin5 siRNA group was given treatment of transfection reagent and Plin5 specific siRNA.In order to further confirm the specific mechanism of Plin5 in high fat/glucose inducing MCMECs injury,MCMECs in Plin5 siRNA group were divided into vehicle group and N-acetyl cysteine (NAC) group,and given the same intervention of high fat.The apoptotic rate was detected by flow cytometry,qRT-PCR and Western blotting were respectively used to detect the mRNA and protein expression of Plin5,and the intracellular reactive oxygen species (ROS) level was tested by DHE staining and ELISA kit.Results The apoptotic rate of MCMECs was increased in a fat concentration-dependent manner (P＜0.05).Compared with 0μmol/L palmitic acid group,the intracellular ROS content and the expression of Plin5 increased significantly in 300μmol/L palmitic acid group (P＜0.05).Compared with the control group and the Scra siRNA group,the intracellular ROS content and apoptotic rate increased significantly in Plin5 siRNA group under the action of 300μmol/L palmitic acid (P＜0.05).Compared with the vehicle group,the intracellular ROS content and apoptotic rate decreased remarkably in NAC group (P＜0.05).Conclusion With inhibition of oxidative stress,Plin5 may reduce the apoptosis of MCMECs induced by high fat and high glucose.
ABSTRACT
Lipolysis is defined as the sequential hydrolysis of triacylglycerol (TAG) stored in cell lipid droplets. For many years, it was believed that hormone-sensitive lipase (HSL) and monoacylglycerol lipase (MGL) were the main enzymes catalyzing lipolysis in the white adipose tissue. Since the discovery of adipose triglyceride lipase (ATGL) in 2004, many studies were performed to investigate and characterize the actions of this lipase, as well as of other proteins and possible regulatory mechanisms involved, which reformulated the concept of lipolysis. Novel findings from these studies include the identification of lipolytic products as signaling molecules regulating important metabolic processes in many non-adipose tissues, unveiling a previously underestimated aspect of lipolysis. Thus, we present here an updated review of concepts and regulation of white adipocyte lipolysis with a special emphasis in its role in metabolism homeostasis and as a source of important signaling molecules.
Subject(s)
Humans , Adipose Tissue, White/enzymology , Lipase/metabolism , Lipolysis/physiology , Adipose Tissue, White/physiology , Lipase/physiologyABSTRACT
[ ABSTRACT] AIM:To observe the changes of perilipin and adipose differentiation-related protein ( ADRP) du-ring the development of diabetes mellitus and to explore the effect of perilipin and ADRP on abnormal glucose metabolism with non-alcoholic fatty liver disease ( NAFLD) .METHODS:The rat model of impaired glucose tolerance ( IGT) was in-duced by feeding high-fat diet, and the type 2 diabetes mellitus (T2DM) model was induced by feeding high-fat diet for 4 weeks and intraperitoneally injecting streptozotocin.The morphological change of the liver tissue was observed under optical microscope.The serum contents of perilipin and ADRP were measured by ELISA.The mRNA expression of perilipin and ADRP in the liver tissues was detected by real-time PCR.The protein expression of ADRP in the liver tissues was deter-mined by Western blotting.RESULTS:HE staining showed steatosis in the liver of the rats in IGT group was more serious than that in T2DM group.The biochemical and the pathological processes of rat models were consistent with the clinical feature of related diseases.The serum content of perilipin had no difference among various groups.The mRNA expression of perilipin in IGT group and T2DM group was significantly higher than that in control group.Compared with IGT group, the mRNA expression of perilipin in T2DM group was significantly increased.The serum content of ADRP in T2DM group was significantly lower than that in control group.The mRNA and protein expression of ADRP in model groups was signifi-cantly lower than that in control group.Compared with IGT group, the mRNA expression of ADRP in T2DM group was sig-nificantly reduced.CONCLUSION: The serum content of ADRP plays a role in the development and progression of T2DM.It is negatively correlated with HOMA-IR.NAFLD occurs during progression of abnormal glucose metabolism in-duced by feeding high-fat diet.The development of abnormal glucose metabolism with NAFLD is probably related to the in-creased expression of perilipin and the reduced expression of ADRP.
ABSTRACT
BACKGROUND: Tumor necrosis factor (TNF)-alpha and AMP-activated protein kinase (AMPK) are known to stimulate and repress lipolysis in adipocytes, respectively; however, the mechanisms regulating these processes have not been completely elucidated. METHODS: The key factors and mechanism of action of TNF-alpha and AMPK in lipolysis were investigated by evaluating perilipin expression and activity of protein kinase RNA-like endoplasmic reticulum kinase (PERK)/eukaryotic initiation factor 2 alpha (eIF2alpha) by Western blot and an immunofluorescence assay in 24-hour TNF-alpha-treated 3T3-L1 adipocytes with artificial manipulation of AMPK activation. RESULTS: Enhancement of AMPK activity by the addition of activator minoimidazole carboxamide ribonucleotide (AICAR) suppressed TNF-alpha-induced lipolysis, whereas the addition of compound C, an inhibitor of AMPK phosphorylation, enhanced lipolysis. Perilipin, a lipid droplet-associated protein, was decreased by TNF-alpha and recovered following treatment with AICAR, showing a correlation with the antilipolytic effect of AICAR. Significant activation of PERK/eIF2alpha, a component of the unfolded protein response signaling pathway, was observed in TNF-alpha or vesicle-treated 3T3-L1 adipocytes. The antilipolytic effect and recovery of perilipin expression by AICAR in TNF-alpha-treated 3T3-L1 adipocytes were significantly diminished by treatment with 2-aminopurine, a specific inhibitor of eIF2alpha. CONCLUSION: These data indicated that AICAR-induced AMPK activation attenuates TNF-alpha-induced lipolysis via preservation of perilipin in 3T3-L1 adipocytes. In addition, PERK/eIF2alpha activity is a novel mechanism of the anti-lipolytic effect of AICAR.