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BACKGROUND@#All-trans retinoic acid (ATRA) promotes the osteogenic differentiation induced by bone morphogenetic protein 9 (BMP9), but the intrinsic relationship between BMP9 and ATRA keeps unknown. Herein, we investigated the effect of Cyp26b1, a critical enzyme of ATRA degradation, on the BMP9-induced osteogenic differentiation in mesenchymal stem cells (MSCs), and unveiled possible mechanism through which BMP9 regulates the expression of Cyp26b1. @*METHODS@#ATRA content was detected with ELISA and HPLC–MS/MS. PCR, Western blot, and histochemical staining were used to assay the osteogenic markers. Fetal limbs culture, cranial defect repair model, and micro–computed tomographic were used to evaluate the quality of bone formation. IP and ChIP assay were used to explore possible mechanism. @*RESULTS@#We found that the protein level of Cyp26b1 was increased with age, whereas the ATRA content decreased. The osteogenic markers induced by BMP9 were increased by inhibiting or silencing Cyp26b1 but reduced by exogenous Cyp26b1. The BMP9-induced bone formation was enhanced by inhibiting Cyp26b1. The cranial defect repair was promoted by BMP9, which was strengthened by silencing Cyp26b1 and reduced by exogenous Cyp26b1. Mechanically, Cyp26b1 was reduced by BMP9, which was enhanced by activating Wnt/b-catenin, and reduced by inhibiting this pathway. b-catenin interacts with Smad1/5/9, and both were recruited at the promoter of Cyp26b1. @*CONCLUSIONS@#Our findings suggested the BMP9-induced osteoblastic differentiation was mediated by activating retinoic acid signalling, viadown-regulating Cyp26b1. Meanwhile, Cyp26b1 may be a novel potential therapeutic target for the treatment of bone-related diseases or accelerating bone-tissue engineering.
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This study aimed to provide a scientific basis for the application of the mycorrhizal planting technology of Dendrobium officinale by investigating the effects of mycorrhizal planting on the fingerprints of D. officinale and the content of six chemical components. Seventeen samples of D. officinale under mycorrhizal and conventional planting were collected from four regions, such as Jinhua of Zhejiang. The HPLC fingerprints were established to evaluate the similarity of the samples. The content of six chemical components of the samples was determined by HPLC. There were 15 common peaks in the fingerprints, and five of them were identified by marker compounds, which were naringenin, 4,4'-dihydroxy-3,5-dimethoxybibenzyl, 3,4'-dihydroxy-5-methoxybibenzyl, 3',4-dihydroxy-3,5'-dimethoxybibenzyl(gigantol), and 3,4-dihydroxy-4',5-dimethoxybibenzyl(DDB-2). The similarities of the fingerprints of mycorrhizal and conventional planting samples and the control fingerprint were in the ranges of 0.733-0.936 and 0.834-0.942, respectively. The influences of mycorrhizal planting on fingerprints were related to planting regions, the germplasm of D. officianle, and the amount of fungal agent. The content of six chemical components in the samples varied greatly, and the content of DDB-2 was the highest, ranging from 69.83 to 488.47 μg·g~(-1). The mycorrhizal planting samples from Chongming of Shanghai and Taizhou of Jiangsu showed an increase in the content of 5-6 components, while samples from Zhangzhou of Fujian and Jinhua of Zhejiang showed an increase in the content of 1-2 components. The results showed that mycorrhizal planting technology did not change the chemical profile of small molecular chemical components of D. officinale, but affected the content of chemical components such as bibenzyls, which has a good application prospect.
Subject(s)
Dendrobium/chemistry , Mycorrhizae , China , Chromatography, High Pressure LiquidABSTRACT
OBJECTIVE@#To analyze factors associated with intestinal acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation(allo-HSCT) in children and to develop a prediction model for intestinal aGVHD after allo-HSCT in children.@*METHODS@#The clinical data of 62 children who underwent allo-HSCT at the Department of Hematology of the People's Hospital of Xinjiang Uygur Autonomous Region from February 2018 to September 2021 were retrospectively analyzed. Intestinal aGVHD was evaluated according to the Mount Sinai Acute GVHD International Consortium (MAGIC) grading criteria, the variables were screened by LASSO (least absolute shrinkage and selection operator) regression analysis with 10-fold cross-validation, and developed a model for predicting intestinal aGVHD after allo-HSCT in children.@*RESULTS@#A total of 33 (53.2%) of the 62 children developed intestinal aGVHD, of which 25 were degree II and 8 were degree III-IV. The results of screening variables by 10-fold cross-validated LASSO regression showed that the significant variables included ethnic minorities (OR =7.229; 95%CI: 2.337-22.354), platelet (PLT) (OR =0.971; 95%CI: 0.932-0.993), uric acid (UA) (OR =0.971; 95%CI: 0.935-0.988), C-reactive protein (CRP) (OR =1.217; 95%CI: 1.053-1.545), and viral infection (OR =10; 95%CI: 3.021-32.668), and these variables were independently associated with intestinal aGVHD in children (all P <0.05). A prediction model was constructed based on above variables. The area under the receiver operating characteristic (ROC) curve (AUC) of the model was calculated, and the AUC value was 0.985 (0.966-1), the Brier score was 0.055. The evaluation showed that the model has a high degree of discrimination and calibration.@*CONCLUSION@#Ethnic minorities, low PLT, low UA, high CRP, and viral infections are independently associated with intestinal aGVHD in children, and early attention should be paid to these high-risk children.
Subject(s)
Humans , Child , Retrospective Studies , Hematopoietic Stem Cell Transplantation/adverse effects , Graft vs Host Disease/diagnosis , Risk Factors , C-Reactive Protein , Acute DiseaseABSTRACT
Hepatic ischemia/reperfusion injury (HIRI) is a serious complication that occurs following shock and/or liver surgery. Gut microbiota and their metabolites are key upstream modulators of development of liver injury. Herein, we investigated the potential contribution of gut microbes to HIRI. Ischemia/reperfusion surgery was performed to establish a murine model of HIRI. 16S rRNA gene sequencing and metabolomics were used for microbial analysis. Transcriptomics and proteomics analysis were employed to study the host cell responses. Our results establish HIRI was significantly increased when surgery occurred in the evening (ZT12, 20:00) when compared with the morning (ZT0, 08:00); however, antibiotic pretreatment reduced this diurnal variation. The abundance of a microbial metabolite 3,4-dihydroxyphenylpropionic acid was significantly higher in ZT0 when compared with ZT12 in the gut and this compound significantly protected mice against HIRI. Furthermore, 3,4-dihydroxyphenylpropionic acid suppressed the macrophage pro-inflammatory response in vivo and in vitro. This metabolite inhibits histone deacetylase activity by reducing its phosphorylation. Histone deacetylase inhibition suppressed macrophage pro-inflammatory activation and diminished the diurnal variation of HIRI. Our findings revealed a novel protective microbial metabolite against HIRI in mice. The potential underlying mechanism was at least in part, via 3,4-dihydroxyphenylpropionic acid-dependent immune regulation and histone deacetylase (HDAC) inhibition in macrophages.
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Oligoasthenoteratozoospermia (OAT) refers to the combination of various sperm abnormalities, including a decreased sperm count, reduced motility, and abnormal sperm morphology. Only a few genetic causes have been shown to be associated with OAT. Herein, we identified a novel homozygous frameshift mutation in meiosis-specific nuclear structural 1 (MNS1; NM_018365: c.603_604insG: p.Lys202Glufs*6) by whole-exome sequencing in an OAT proband from a consanguineous Chinese family. Subsequent variant screening identified four additional heterozygous MNS1 variants in 6/219 infertile individuals with oligoasthenospermia, but no MNS1 variants were observed among 223 fertile controls. Immunostaining analysis showed MNS1 to be normally located in the whole-sperm flagella, but was absent in the proband's sperm. Expression analysis by Western blot also confirmed that MNS1 was absent in the proband's sperm. Abnormal flagellum morphology and ultrastructural disturbances in outer doublet microtubules were observed in the proband's sperm. A total of three intracytoplasmic sperm injection cycles were carried out for the proband's wife, but they all failed to lead to a successful pregnancy. Overall, this is the first study to report a loss-of-function mutation in MNS1 causing OAT in a Han Chinese patient.
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Objective:To explore the relationship between fragmented QRS complex and heart rate variability (HRV) and ventricular arrhythmia in patients with old myocardial infarction.Methods:From August 2018 to October 2019, 200 patients with old myocardial infarction were first treated in the Department of cardiac function examination of the First Affiliated Hospital of Hebei North University. The patients were divided into 99 cases of old myocardial infarction with fragmented QRS wave group and 101 cases of old myocardial infarction without fragmented QRS wave group according to the case bank data and conventional 12 lead ECG diagnosis in our hospital for the first time. Then, the 24-h ambulatory ECG reexamined within 1 year after discharge was retrospectively analyzed. The incidence of ventricular arrhythmia was compared between the two groups by χ 2 test. The difference of heart rate variability between the two groups was compared by rank sum test. Multiple logistic regression was used to analyze the value of different indexes of heart rate variability in the evaluation of fragmented QRS complex in old myocardial infarction. Drawing the receiver operating characteristic (ROC), and the area under the curve (AUC) was used to analyze the diagnostic accuracy of different indexes of heart rate variability in the broken QRS complex of old myocardial infarction. Results:According to the Lown classification of ventricular premature contraction, the number of positive ventricular arrhythmias in patients with Grade Ⅰ of ventricular premature contraction and Grade Ⅲ-Ⅴ of ventricular premature contraction in the old myocardial infarction fragmented QRS group was higher than that in the old myocardial infarction non fragmented QRS group (Grade Ⅰ of ventricular premature contraction: 54.5% (54/99)and 39.6%(40/101); χ 2=4.484, P<0.05;Grade Ⅲ-Ⅴ of ventricular premature contraction: 34.3% (34/99) and 9.9%(10/101); χ 2=17.406, P<0.05)). Ventricular premature contraction Grade 0 old myocardial infarction fragmented QRS group was lower than old myocardial infarction non fragmented QRS group (8.1% (8/99) and 48.5% (49/101); χ 2=37.995, P<0.05). The total number of positive cases of ventricular arrhythmia in the old myocardial infarction group with fragmented QRS wave was higher than that in the old myocardial infarction group without fragmented QRS wave (91.9% (91/99) and 51.5%(52/101); χ 2=57.146, P<0.05)). There was no significant difference in the number of positive ventricular arrhythmias between the old myocardial infarction fragmentation QRS group and the old myocardial infarction non fragmentation QRS group ( P>0.05). The standard deviation of NN intervals (SDNN) and the standard deviation of average NN intervals (SDANN) of HRV time domain indexes in the old myocardial infarction fragmented QRS group were higher than those in the old myocardial infarction non fragmented QRS Group (SDNN:143.00(122.00,166.00) vs. 110.00(95.00,130.50), Z=5.780, P<0.05; SDANN:112.00(100.00,136.00) vs. 96.00(76.00,118.50), Z=4.013, P<0.05). Multiple Logistics regression analysis results of HRV domain shows that HRV time domain SDNN and SDANN have diagnositic value in diagnosis fQRS after OMI(SDNN: OR=0.949, 95% CI:0.922-0.977, P<0.001; SDANN: OR=1.036, 95% CI:1.005-1.068, P=0.022). Area under ROC curve of HRV time domain SDNN and SDANN have particular diagnositic accuracy in diagnosis fQRS after OMI(SDNN: AUC 0.737, 95% CI 0.666-0.807, Sensitivity 0.818, Specificity 0.634; SDANN: AUC 0.664, 95% CI 0.587-0.741, Sensitivity 0.737, Specificity 0.673. 0.5<AUC<1). Conclusion:Fragmented QRS complex was positively correlated with the incidence and severity of ventricular arrhythmia in patients with old myocardial infarction, and positively correlated with time-domain indexes SDNN and SDANN of heart rate variability in patients with old myocardial infarction.
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Exogenous calcium can enhance the resistance of certain plants to abiotic stress. However,the role of calcium insaltstressed honeysuckle is unclear. The study is aimed to investigate the effects of exogenous calcium on the biomass,chlorophyll content,gas exchange parameters and chlorophyll fluorescence of honeysuckle under salt stress. The results showed that the calcium-treated honeysuckle had better photochemical properties than the salt-stressed honeysuckle,such as PIABS,PItotal,which represents the overall activity of photosystemⅡ(PSⅡ),and related parameters for characterizing electron transport efficiency φP0,ψE0,φE0,σR,and φR are significantly improved. At the same time,the gas exchange parameters Gs,Ci,Trare also maintained at a high level. In summary,exogenous calcium protects the activity of PSⅡ,promotes the transmission of photosynthetic electrons,and maintains a high Ci,therefore enhances the resistance of honeysuckle under salt stress.
Subject(s)
Calcium , Pharmacology , Chlorophyll , Lonicera , Physiology , Photosynthesis , Plant Leaves , Salt StressABSTRACT
The purpose of the present study was to investigate the effect of advanced glycated albumin (AGE-alb) on pyroptosis of macrophages and the underlying molecular mechanisms. RAW264.7 macrophages were treated with AGE-alb (1, 2, 4 and 6 g/L) and control albumin (C-alb, 4 g/L) for 24 h, or preincubated with MCC950 (1 μmol/L) for 1 h and then treated with AGE-alb (4 g/L) for 24 h. Cell viability and caspase-1 activity were measured by MTT and assay kits, respectively. Lactate dehydrogenase (LDH) activity and the levels of interleukin-1β (IL-1β) and IL-18 in media were detected. Cell death degree was evaluated by TUNEL and Hoechst 33342/PI staining. The protein levels of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), procaspase-1 and cleaved caspase-1 were assessed by Western blot. The results showed that AGE-alb treatment caused obvious decrease in cell viability and increases in LDH leakage and the percentages of TUNEL- or PI-positive cells in a concentration-dependent manner. Additionally, AGE-alb promoted IL-1β and IL-18 secretion, upregulated NLRP3 expression, and increased caspase-1 activity especially at the dose of 4 and 6 g/L. However, MCC950 (an NLRP3 inhibitor) pretreatment inhibited significantly the decrease in cell viability and the increases in LDH leakage and percentages of TUNEL- or PI-positive cells induced by AGE-alb. Furthermore, MCC950 attenuated obviously AGE-alb-induced IL-1β and IL-18 secretion and caspase-1 activation. These results indicate that AGE-alb may induce macrophage pyroptosis, and the mechanism is at least partially by activating NLRP3-caspase-1 pathway.
Subject(s)
Caspase 1 , Gene Expression Regulation , Interleukin-1beta , Genetics , Macrophages , NLR Family, Pyrin Domain-Containing 3 Protein , Genetics , Pyroptosis , Serum Albumin , PharmacologyABSTRACT
Asthma is a common inflammatory disease in childhood. The interaction of genes, environment and host factors contributes to the development and progression of asthma, and the heterogeneity of asthma leads to the complexity of asthma phenotype classification. In this article, we reviewed recent analyses of asthma phenotypes in children using different criteria, including the age of onset, breathing pattern, clinical characteristics, airway inflammatory cells, risk factors, immunity and genetics, and assessed the prognosis of previous asthma phenotypes based on the above criteria, hoping to provide evidence for the treatment and prognosis of children with asthma.
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The purpose of this study was to investigate whether activating transcription factor 6 (ATF6), a sensor to endoplasmic reticulum stress (ERS), would mediate advanced glycated albumin (AGE-alb)-induced macrophage apoptosis and to elucidate the possible molecular mechanisms. RAW264.7 macrophages were cultured in vitro and treated with AGE-alb (2, 4 and 6 g/L), normal control albumin or tunicamycin (TM, 4 mg/L) for 24 h. ATF6 small interfering RNA (siRNA) was transfected to RAW264.7 cells by Lipofectamine 2000. Cell viability and apoptosis were determined by MTT method and Annexin V-FITC/propidium iodide apoptosis detection kit, respectively. The activities of lactate dehydrogenase (LDH) in medium and caspase-3 in cells were measured by corresponding detection kits. ATF6 nuclear translocation was detected by Western blot and immunofluorescence cytochemistry. Protein and mRNA levels of C/EBP homologous protein (CHOP, a key-signaling component of ERS-induced apoptosis) were detected by Western blot and real-time fluorescence quantitative PCR, respectively. The results showed that similar to TM, AGE-alb increased the expression of CHOP at both the protein and mRNA levels in a concentration dependent manner. ATF6, as a factor that positively regulates CHOP expression, was activated by AGE-alb in a concentration dependent manner. siRNA-mediated knockdown of ATF6 significantly inhibited AGE-alb-induced macrophage injury, as indicated by the increased cell viability and the decreased LDH release, apoptosis and caspase-3 activation. Additionally, ATF6 siRNA attenuated AGE-alb-induced CHOP upregulation at both the protein and mRNA levels. These results suggest that ATF6 and its downstream molecule CHOP are involved in AGE-alb-induced macrophage apoptosis.
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Autophagy is a cellular catabolic process responsible for removing the injured proteins and organelles via lysosome-dependent pathway, and it plays an important role in maintaining cellular homeostasis. Recent studies have shown that autophagy is activated and implicated in the pathogenesis of atherosclerosis. Autophagy can be triggered by oxidative lipids, cytokines and advanced glycation end products, and exerts protective or detrimental functions in the progression of atherosclerosis. However, the precise role and mechanisms of autophagy in different stages of atherosclerosis are still not fully clarified. This review highlights recent findings regarding autophagy response in vascular cells and its potential contribution to atherogenesis. Additionally, the relationship of autophagy with endoplasmic reticulum stress and whether autophagy could be a new therapeutic target for atherosclerosis are also discussed.
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The purpose of the present study was to investigate the effect of advanced glycated albumin (AGE-alb) on the activation of caspase-12, a key molecule in endoplasmic reticulum stress (ERS)-associated apoptotic pathway, and to elucidate the underlying molecular mechanisms of macrophage apoptosis. RAW264.7 macrophages were treated with AGE-alb (2, 4 and 6 g/L), control albumin (C-alb, 4 g/L), tunicamycin (TM, 4 mg/L), or pretreated with 4-phenylbutyric acid (PBA, 5 mmol/L) for 1 h and then treated with AGE-alb (4 g/L). After incubation for 24 h, the cell viability and apoptosis were determined by using MTT assay and TUNEL detection kit, respectively. Lactate dehydrogenase (LDH) activity in media was determined by using an assay kit. The protein levels of caspase-12 were examined by Western blot analysis. The results showed that like TM (an ERS inducer), incubation with AGE-alb led to significant decrease in viability and increase in LDH activity in media and apoptotic rate in a dose-dependent manner. In addition, AGE-alb induced activation of caspase-12 especially at the concentration of 4 and 6 g/L (P < 0.01), which was similar to TM. However, PBA (an ERS inhibitor) protected RAW264.7 macrophages from AGE-alb-induced decrease in viability and increases in LDH activity and apoptosis. Moreover, PBA also inhibited the caspase-12 activation induced by AGE-alb (P < 0.05). These results suggest that AGE-alb may induce apoptosis in RAW 264.7 macrophages, and the mechanism may be related to the activation of ERS-associated apoptotic pathway mediated by caspase-12.
Subject(s)
Animals , Mice , Apoptosis , Caspase 12 , Cell Line, Tumor , Cell Survival , Endoplasmic Reticulum Stress , Macrophages , Phenylbutyrates , Serum Albumin , TunicamycinABSTRACT
Pigment epithelium-derived factor (PEDF) is a multifunctional protein with anti-inflammatory, antioxidant and antithrombotic properties and plays a protective role against atherosclerosis (AS). The purpose of the present study is to explore the effects of oxidized low density lipoprotein (ox-LDL) on the expression of PEDF in cultured human umbilical vein endothelial cells (HUVECs). HUVECs were cultured and incubated with ox-LDL at different concentrations (6.25, 12.5, 25, 50, 100 and 150 mg/L) for 24 h. Apoptosis of endothelial cells were assayed by morphological staining and flow cytometry. The intracellular reactive oxygen species (ROS) levels were measured by flow cytometry. Cell viability was assayed by MTT assay. PEDF protein and mRNA expressions in HUVECs were analyzed by Western blot and quantitative real-time PCR, respectively. The results showed that ox-LDL significantly induced apoptosis, reduced cell viability, increased intracellular ROS levels and decreased the PEDF expression in HUVECs in a concentration-dependent manner. Ox-LDL at 50 mg/L obviously decreased the PEDF protein expression compared with control group (P < 0.05), whereas 25 mg/L ox-LDL already markedly reduced the PEDF mRNA expression (P < 0.05). In conclusion, the results suggest that ox-LDL down-regulates the PEDF expression through an increased ox-LDL-induced intracellular production of ROS.
Subject(s)
Humans , Apoptosis , Cells, Cultured , Down-Regulation , Eye Proteins , Metabolism , Human Umbilical Vein Endothelial Cells , Cell Biology , Lipoproteins, LDL , Pharmacology , Nerve Growth Factors , Metabolism , Reactive Oxygen Species , Metabolism , Serpins , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the efficacy of hematopoietic stem cell (HSC) transplantation via the hepatic artery vs. the portal vein for end-stage liver disease (ESLD).</p><p><b>METHODS</b>Patients with hepatic decompensation were prospectively recruited from September 2010 to September 2012 to receive HSC transplantation via the hepatic artery or the portal vein. Liver function was examined at 3, 6, and 12 months after transplantation. Liver biopsy Results were analyzed using the Knodell score.</p><p><b>RESULTS</b>Eighty patients (58 males and 22 females) were enrolled in the study. The Child-Pugh score was grade B in 69 cases, and grade C in the remaining 11 cases. HSC transplantation was performed via the portal vein in 36 patients and via the hepatic artery in 44 patients. ALT levels decreased while serum albumin levels increased significantly in both groups at 6 and 12 months after HSC transplantation (P<0.05 compared with pre-transplantation levels). Total bilirubin levels decreased significantly in both groups at 3, 6, and 12 months after HSC transplantation (P<0.05 compared with pre-transplantation levels). Additionally, prothrombin time decreased in both groups at 12 months after HSC transplantation (P<0.05 compared with pre-transplantation level). There were no significant differences in ALT, total bilirubin and prothrombin time between the two groups either before or after transplantation. Moreover, Knodell score decreased significantly at 6 and 12 months. Histological examination showed that liver cell edema, degeneration, necrosis, and inflammation were significantly relieved at 3, 6, and 12 months after transplantation. The incidence of portal vein thrombosis, upper gastrointestinal bleeding, and hepatic encephalopathy were 1.25%, 3.75%, and 2.5% respectively. The one-year survival rate was 100%.</p><p><b>CONCLUSIONS</b>Autologous HSC transplantation improves liver function and histology in ESLD patients. The administration route of HSC has no significant impact on the efficacy of transplantation.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Disease-Free Survival , End Stage Liver Disease , Pathology , Therapeutics , Hematopoietic Stem Cell Transplantation , Methods , Hepatic Artery , Infusions, Intra-Arterial , Infusions, Intravenous , Liver Function Tests , Portal Vein , Prospective Studies , Treatment OutcomeABSTRACT
The present study was to investigate whether endoplasmic reticulum stress (ERS) was involved in oxidized low density lipoprotein (ox-LDL)-induced scavenger receptor A1 (SR-A1) upregulation in macrophages. RAW264.7 cells were pretreated with 20 mmol/L of 4-phenylbutyric acid (PBA) for 30 min and then treated with ox-LDL (50 mg/L) for 12 h or stimulated with 2 mg/L tunicamycin (TM) or 2 μmol/L thapsigagin (TG) for 4 h. In addition, RAW264.7 cells were incubated with 0.5, 1 and 2 mg/L TM for 4 h or treated with 2 mg/L TM for 1, 2 and 4 h, respectively. The intracellular total cholesterol (TC) content was measured using a tissue/cell total cholesterol assay kit. The protein and mRNA expressions of SR-A1 and glucose-regulated protein 78 (GRP78) were analyzed by Western blot and real-time PCR, respectively. Dil-ox-LDL uptake was detected using a microplate reader. The results showed that ox-LDL-induced cholesterol accumulation in macrophages was attenuated by PBA, an ERS inhibitor. Ox-LDL caused significant SR-A1 upregulation with concomitant activation of ERS as assessed by upregulation of GRP78, whereas PBA significantly inhibited the ox-LDL-induced SR-A1 upregulation (P < 0.05) and slightly decreased GRP78 expression by 39.3% (P = 0.057). TM, an ERS inducer, upregulated SR-A1 protein expression and ox-LDL uptake in dose- and time-dependent manner, but had no significant effect on SR-A1 mRNA level. However, the TM- or TG-induced SR-A1 upregulation and ox-LDL uptake were significantly mitigated by PBA. These data indicate that ERS plays a critical role in ox-LDL-induced SR-A1 upregulation, which in turn enhances the foam cell formation by uptaking more ox-LDL.
Subject(s)
Animals , Mice , Cell Line , Cholesterol , Metabolism , Endoplasmic Reticulum Stress , Heat-Shock Proteins , Metabolism , Lipoproteins, LDL , Pharmacology , Macrophages , Metabolism , Scavenger Receptors, Class A , Metabolism , Up-RegulationABSTRACT
<p><b>OBJECTIVE</b>To explore an optimal anesthesia method with less impact on hemodynamics and electrocardiogram (ECG) of old patients with coronary artery disease (CHD) during abdominal operation.</p><p><b>METHODS</b>The 133 CHD patients waiting for abdominal operation were randomly divided into continuous epidural anesthesia (EA) group, general anesthesia group (GA) and combined spinal-epidural anesthesia (CSEA) group. Continuous monitoring was carried out during operation and mean arterial pressure (MAP), heart rate (HR), oxygen saturation (SaO2), abnormal ECG were measured respectively at different time for comparison and the differences of the above hemodynamic parameters and abnormal ECG features were compared among the 3 groups.</p><p><b>RESULTS</b>At the 15 min and 30 min point after anesthesia, SaO2 in GA group was significantly increased compared to that in the EA group (P < 0.05). At 15 min, 30 min and 60 min point after anesthesia, MAP in CSEA group was significantly increased compared to that in the EA group (P < 0.05). At 30 min point after anesthesia, HR in CSEA group was increased significantly compared to the EA group (P < 0.05). At 15 min and 30 min point after anesthesia, SaO2 in the CSEA group was increased significantly compare to the EA group (P < 0.05). Compared with preanesthesia (T0) in EA group, MAP, HR and SaO2 decreased significantly at 15, 30 and 60 min after anesthesia (P < 0.05). The fluctuation of the three parameters in GA and CSEA groups were relatively small (P > 0.05). As well as the comparison of abnormal ECG among the 3 groups was concerned, the incidence of ST-T changes in GA and CSEA groups were significantly lower than that in EA group at the time of 15 min, 30 min and 60 min after anesthesia and at the time of surgery termination (P < 0.05, P < 0.01). The incidence of arrhythmia in GA and CSEA groups were significantly lower than that in EA group at the time of 15 min, 30 min and 60 min after anesthesia (P < 0.05, P < 0.01). Compared with T0 in the same group, the incidences of ST-T changes and arrhythmia in GA or CSEA group at the time of 15, 30 and 60 min after anesthesia and at the time of surgery termination were significantly lower than that before anesthesia (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>GA and CSEA have less impact on hemodynamics and have smaller incidence of abnormal ECG of old CHD patients with abdominal operation.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Anesthesia , Methods , Coronary Disease , Electrocardiography , Hemodynamics , Perioperative PeriodABSTRACT
<p><b>OBJECTIVE</b>To explore the perioperative cardiovascular dysfunction and its relevance to age in patients with silent coronary heart disease (or silent myocardial ischemia), and explore the clinical treatment and recovery of perioperative arrhythmias.</p><p><b>METHODS</b>One hundred and eighty cases were selected from selective surgery patients with silent myocardial ischemia (SMI). Among the cases, 130 patients older than 51 years old were divided into 51 - 60 year-old group, 61- 70 year-old group and 71 - 80 year-old group. Control group was set up by other 50 patients younger than 51 years old. Electrocardiogram data of 24 h before the operation, 24 h after the operation and 48 h after the operation were continuously monitored by dynamic electrocardiogram (DCG). The electrocardiogram data of ST shifting, arrhythmia incidences of different type and at different time were analyzed by professional doctors. At the same time, the treatment and recovery of perioperative arrhythmia were recorded.</p><p><b>RESULTS</b>As the age increase, the magnitude and duration of ST shifting appeared upward trend compared to the control group (P < 0.05, P < 0.01). The incidence of ST elevation in 71 - 80 year-old group was higher than the control group (P < 0.05). The ST depression duration in 61 - 70 and 71 - 80 year-old group and ST elevation magnitude in 71 - 80 year-old group were higher than 51 - 60 year-old group (P < 0.05). Compared to the control group, the incidence of accelerated idioventricular rhythm (AIR) in 61 - 70 year-old group and the incidence of sinus bradycardia (SB), ventricular premature beat (VPB), ventricular tachycardia (VT) in 71 - 80 year-old group were higher (P < 0.05, P < 0.01). Compared to the 51 - 60 year-old group, the incidence of atrial fibrillation (AF) in 61 - 70 year-old group and the incidence of VP, VT, AF in 71 - 80 year-old group were higher (P < 0.05, P < 0.01). The arrhythmia incidences in 24 h after operation were higher than 48 h after operation and 24 h before operation (P < 0.01). As the age increase, the recovery incidence by removing inducement was decreased, but the recovery incidences by drug and electric-shock treatment were increased (P < 0.05).</p><p><b>CONCLUSION</b>Old SMI patients have high levels of perioperative myocardial ischemia and arrhythmia, and 24 h after operation is the period of high incidence.</p>
Subject(s)
Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Cardiovascular System , Coronary Disease , Perioperative PeriodABSTRACT
The purposes of the present study were to investigate the inhibitory effect of quercetin (QUE) preconditioning on endoplasmic reticulum stress (ERS) inducer tunicamycin (TM)-induced apoptosis in RAW264.7 macrophages and the underlying molecular mechanisms. RAW264.7 cells were pretreated with different concentrations (20, 40, and 80 μmol/L) of QUE for 30 min and then treated with TM (5 mg/L) for 12 h. Cell viability and apoptosis were determined using MTT assay and Annexin V-FITC apoptosis detection kit, respectively. The nuclear translocation of activating transcription factor 6 (ATF6) in cells was detected by immunofluorescence analysis and Western blot. Protein and mRNA expressions of C/EBP homologous protein (CHOP) and Bcl-2 were examined by Western blot and real-time PCR, respectively. The results showed that TM reduced cell viability and induced apoptosis in RAW264.7 macrophages. The cytotoxic effects of TM were significantly inhibited by QUE pretreatment at the concentrations of 40 and 80 μmol/L. Interestingly, we found that QUE also significantly suppressed the TM-induced translocation of ATF6, an ERS sensor, from the cytoplasm to the nucleus. In addition, exposure of RAW264.7 macrophages to TM resulted in a significant increase of the expression of CHOP, a transcription factor regulated by ATF6 under conditions of ERS, as well as a decrease of Bcl-2 at transcript and protein levels. QUE blocked these effects in a dose-dependent manner. These data indicate that QUE can protect RAW264.7 cells from TM-induced apoptosis and that the mechanism at least partially involves its ability to inhibit the ATF6-CHOP signaling pathway.
Subject(s)
Animals , Mice , Activating Transcription Factor 6 , Metabolism , Apoptosis , Cell Survival , Endoplasmic Reticulum Stress , Macrophages , Cell Biology , Quercetin , Pharmacology , Transcription Factor CHOP , Metabolism , Tunicamycin , PharmacologyABSTRACT
Endoplasmic reticulum (ER) stress occurs in macrophage-rich areas of advanced atherosclerotic lesions and contributes to macrophage apoptosis and subsequent plaque necrosis. The purpose of the present study was to investigate the effects of caveolin-1 (Cav-1) on ER stress-induced apoptosis in cultured macrophages and the underlying mechanisms. RAW264.7 cells were incubated with thapsigargin (TG) to establish ER stress model. And Cav-1 expression was detected by Western blot. After being pretreated with filipin(III), a caveolae inhibitor, RAW264.7 cells were assayed with flow cytometry and confocal laser scanning microscopy to detect cell apoptosis. Moreover, p38 mitogen-activated protein kinase (MAPK) phosphorylation and C/EBP homologous protein (CHOP) expression were detected with Western blot. The results showed that Cav-1 expression was markedly increased at early stage of TG treatment (P < 0.05) and then decreased with prolonged or high dose TG treatments. The increasing of Cav-1 expression induced by TG in RAW264.7 cells was abolished under inhibition of caveolae by filipin(III) (P < 0.05). The effect of TG on apoptosis of RAW264.7 cells was further augmented after pretreatment with filipin(III) (P < 0.05). Western blotting showed that MAPK phosphorylation induced by TG was inhibited by filipin(III) in RAW264.7 cells (P < 0.05), whereas CHOP remained unchanged (P > 0.05). These results suggest that Cav-1 may play a critical role in suppressing ER stress-induced macrophages apoptosis in vitro, and one of the mechanisms may be correlated with the activation of p38 MAPK prosurvival pathway.
Subject(s)
Animals , Mice , Apoptosis , Caveolin 1 , Genetics , Metabolism , Cell Line , Endoplasmic Reticulum Stress , Physiology , Filipin , Pharmacology , MAP Kinase Signaling System , Macrophages , Cell Biology , Thapsigargin , Pharmacology , Transcription Factor CHOP , Metabolism , p38 Mitogen-Activated Protein Kinases , MetabolismABSTRACT
<p><b>BACKGROUND</b>Ischemic postconditioning (I-postC) is a newly discovered and more amenable cardioprotective strategy capable of protecting the myocardium from ischemia/reperfusion (I/R) injury. Endoplasmic reticulum (ER) is a principal site for secretary protein synthesis and calcium storage. Myocardial I/R causes ER stress and emerging studies suggest that the cardioprotection has been linked to the modulation of ER stress. The aim of the present study was to determine whether cardioprotection of I-postC involves reduction in ER stress through calcineurin pathway.</p><p><b>METHODS</b>In the in vivo model of rat myocardial I/R, myocardial infarct size was measured by triphenyltetrazolium chloride (TTC) staining and apoptosis was detected using terminal eoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) assay. Expression of calreticulin, C/EBP homologous protein (CHOP), caspase-12, and activation of caspase-12 in myocardium were detected by Western blotting. The activity and expression of calcineurin in myocardium were also detected.</p><p><b>RESULTS</b>I-postC protected the I/R heart against apoptosis, myocardial infarction, and leakage of lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB). I-postC suppressed I/R-induced ER stress, as shown by a decrease in the expression of calreticulin and CHOP, and caspase-12 activation. I-postC downregulated calcineurin activation in myocardium subjected to I/R.</p><p><b>CONCLUSION</b>I-postC protects myocardium from I/R injury by suppressing ER stress and calcineurin pathways are not associated with the I-postC-induced suppression of ER stress-related apoptosis.</p>