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Context: Bullous pemphigoid (BP) and “Pemphigus diseases” (PD) can have overlapping clinical manifestations and accurate distinction is crucial for appropriate management. Aims: The study aimed at analyzing the utility of simple hematological markers of systemic inflammation like neutrophil-to-lymphocyte ratio (NLR), neutrophil-to-eosinophil ratio (NER), and platelet-to-lymphocyte ratio (PLR) in clinical decision making in the setting of clinical differentials of BP and PD in a particular case. Methods: This single-centre based retrospective observational analytical study included adult subjects newly diagnosed to have BP (n=66) or PD (n=53), confirmed with direct immune-fluorescence testing, over a period of six years. Blood counts performed using Coulter™ hematology analyser, at the time of their initial presentation, were retrieved from the hospital medical records, and the leucocyte ratios were calculated.Statistical Analysis: The data were compared between the two groups, using Mann–Whitney U test and chi-square test /Fisher's exact test. ROC curve analysis was performed to estimate cut-off values. Results: The BP group had a significantly higher NLR, total leukocyte counts (TLC), absolute eosinophil counts (AEC), and absolute lymphocyte counts (ALC), and lower NER values compared to the PD group (P < 0.05). Areas under ROC for NLR, NER, TLC, AEC, and ALC were between 0.5 and 0.7. NLR ? 7, AEC ? 2055/cumm, and TLC ? 15,000/cumm had a specificity of 90.6, 100, and 100% respectively for identifying BP patients out of the two groups, but with a low sensitivity of 22.7, 21, and 22.7%, respectively. Conclusions: NLR can be a valuable diagnostic adjunct in subtyping autoimmune bullous disorders, albeit in a small proportion of cases.
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Purpose: To observe the effect of puerarin on renal ischemia-reperfusion (I/R) injury in rats, and to explore its mechanism based on NLRP3/Caspase-1/GSDMD pathway. Methods: Twenty-one Sprague-Dawley rats were divided into three groups: sham-operated group (sham), model group (RIRI), and puerarin treatment group (RIRI + Pue). The model of acute renal I/R injury was established by cutting the right kidney and clamping the left renal pedicle for 45 min. Results: Renal function parameters were statistically significant in group comparisons. The renal tissue structure of rats in sham group was basically normal. Pathological changes were observed in the RIRI group. The renal pathological damage score and apoptosis rate in the RIRI group were higher than those in the sham group, and significantly lower in the RIRI + Pue group than in the RIRI group. Indicators of oxidative stress-superoxide dismutase, malondialdehyde, and glutathione peroxidase-were statistically significant in group comparisons. Compared with the sham group, the relative expressions of NLRP3, Caspase-1 and GSDMD proteins in the RIRI group were increased. Compared with the RIRI group, the RIRI + Pue group had significant reductions. Conclusions: Puerarin can inhibit the activation of NLRP3/Caspase-1/GSDMD pathway, inhibit inflammatory response and pyroptosis, and enhance the antioxidant capacity of kidney, thereby protecting renal I/R injury in rats.
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Animals , Rats , Reperfusion Injury , Pyroptosis , Inflammation , Kidney/injuriesABSTRACT
Purpose: Myocardial ischemia/reperfusion injury (MIRI) leads to myocardial tissue necrosis, which will increase the size of myocardial infarction. The study examined the protective effect and mechanism of the Guanxin Danshen formula (GXDSF) on MIRI in rats. Methods: MIRI model was performed in rats; rat H9C2 cardiomyocytes were hypoxia-reoxygenated to establish a cell injury model. Results: The GXDSF significantly reduced myocardial ischemia area, reduced myocardial structural injury, decreased the levels of interleukin (IL-1ß, IL-6) in serum, decreased the activity of myocardial enzymes, increased the activity of superoxide dismutase (SOD), and reduced glutathione in rats with MIRI. The GXDSF can reduce the expression of nucleotide- binding oligomerization domain, leucine-rich repeat and pyrin domain containing nod-like receptor family protein 3 (NLRP3), IL-1ß, caspase-1, and gasdermin D (GSDMD) in myocardial tissue cells. Salvianolic acid B and notoginsenoside R1 protected H9C2 cardiomyocytes from hypoxia and reoxygenation injury and reduced the levels of tumor necrosis factor α (TNF-α) and IL-6 in the cell supernatant, decreasing the NLRP3, IL-18, IL-1ß, caspase-1, and GSDMD expression in H9C2 cardiomyocytes. GXDSF can reduce the myocardial infarction area and alleviate the damage to myocardial structure in rats with MIRI, which may be related to the regulation of the NLRP3. Conclusion: GXDSF reduces MIRI in rat myocardial infarction injury, improves structural damage in myocardial ischemia injury, and reduces myocardial tissue inflammation and oxidative stress by lowering inflammatory factors and controlling focal cell death signaling pathways.
Subject(s)
Animals , Rats , Myocardial Reperfusion , Reperfusion Injury , Ginsenosides/administration & dosage , NLR Family, Pyrin Domain-Containing 3 ProteinABSTRACT
Objective:To explore the potential role of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)/GATA-binding protein 4 (GATA-4)/vascular endothelial growth factor (VEGF) signal pathway in neovascular age-related macular degeneration (nAMD).Methods:We applied the TRANSFAC Public database to search the human and mouse VEGF promoters and upstream transcription factors, analyzed the transcription factors that may influence the transcriptional activity of VEGF. The RAW264.7 cells were divided into control group and lipopolysaccharide (LPS) stimulated group (LPS group). Real time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the activation of NLRP3 inflammasome, and the mRNA levels of GATA-4 and VEGFA. Thus, we applied the specific small molecular NLRP3 inhibitor MCC950 pretreated RAW264.7 cells (LPS+ MCC950 group), and detected the gene expression of NLRP3, Caspase-1, interleukin 1β( IL-1β), GATA-4 and VEGFA.Results:There were multiple GATA transcription factor binding sites upstream of human and mouse VEGF promoters. Compared with the control group, mRNA expression of NLRP3, Caspase-1, IL-1β, GATA-4 and VEGFA in LPS group were increased (all P<0.05). Compared with LPS group, mRNA expression of NLRP3, Caspase-1, IL-1β, GATA-4 and VEGFA in LPS+ MCC950 group were significantly decreased (all P<0.05). Conclusions:NLRP3/GATA-4/VEGF signal pathway may play a significant role in the pathologic processes of nAMD.
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In recent years, with the development of metabolic reprogramming research, people have changed their understanding of the biological effects of immune cells. Under the stimulation of inflammatory response, immune cells re-regulate their metabolism and bioenergetics, provide energy and substrates for cell survival, and initiate immune effect functions. Nod-like receptor protein 3 (NLRP3) inflammasome, as an important component of the innate immune system, has been shown to sense metabolites such as uric acid and cholesterol crystals, and can be inhibited by metabolites such as ketones. It is also regulated by mitochondrial reactive oxygen species and glycolytic components (such as hexokinase). Recent studies have shown that a variety of metabolic pathways converge as effective regulators of NLRP3 inflammasome. The paper reviews the metabolic regulatory pathways and specificity of NLRP3 inflammasome activation, and its role in renal diseases.
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Objective:To investigate the potential function and related mechanism of microRNA-223 (miRNA-223) in the podocyte pyroptosis of hepatitis B virus (HBV)-associated glomerulonephritis induced by HBV X protein (HBx).Methods:HBx-overexpressing lentivirus was transfected into human renal podocytes to mimic the pathogenesis of HBV-GN. Real-time fluorescence quantitative PCR and Western blotting experiments were used to detect the mRNA and protein expression of pyroptosis-related proteins [nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC) and caspase-1], and inflammatory factors (interleukin-1β and interleukin-18), respectively.TUNEL staining and flow cytometry were used to detect the number of pyroptosis cells. Immunofluorescence staining was used to detect the expression of podocytes biomarkers desmin and nephrin; Hoechst 33342 staining was used to observe the morphological and quantitative changes of podocyte nuclei. Enzyme-linked immunosorbent assay was used to measure caspase-1 activity. The dual luciferase reporter gene assay was used to verify the downstream target of miRNA-223. Podocytes were divided into the following nine groups: control group (no special treatment), empty plasmid group (transfected with empty plasmid), HBx overexpression group (transfected with HBx overexpression lentivirus), HBx overexpression+miRNA-223 mimic group (transfected with HBx overexpression lentivirus and miRNA-223 mimic), HBx overexpression+miRNA-223 inhibitor group (transfected with HBx overexpression lentivirus and miRNA-223 inhibitor), HBx overexpression+miRNA-223 mimic+NLRP3 group (transfected with HBx overexpression lentivirus, miRNA-223 mimic and NLRP3 overexpression plasmid), HBx overexpression+miRNA-223 mimic+ NLRP3 siRNA group (transfected with HBx overexpression lentivirus, miRNA-223 mimic and NLRP3 siRNA), HBx overexpression+miRNA-223 inhibitor+NLRP3 group (transfected with HBx overexpression lentivirus, miRNA-223 inhibitor and NLRP3 overexpression plasmid), HBx overexpression+miRNA-223 inhibitor+NLRP3 siRNA group (transfected with HBx overexpression lentivirus, miRNA-223 inhibitor and NLRP3 siRNA).Results:miRNA-223 was down-regulated in HBx overexpression group compared with the control group ( P < 0.05). TUNEL and immunofluorescence staining showed that NLRP3 knockdown attenuated podocyte injury and pyroptosis induced by HBx overexpression ( P < 0.05). Dual luciferase reporter gene assay demonstrated that NLRP3 was one of the downstream targets of miRNA-223. Rescue experiments revealed that NLRP3 overexpression weakened the protective effect of miRNA-223 in podocyte injury ( P < 0.05). The addition of miRNA-223 mimic and NLRP3 siRNA decreased the expression of NLRP3 inflammasome and cytokines, and reduced the number of pyroptosis cells induced by HBx overexpression (all P < 0.05); The addition of miRNA-223 inhibitor and NLRP3 overexpression plasmid significantly increased the expression of NLRP3 inflammasome and cytokines, caspase-1 activity, and the number of pyroptosis cells (all P < 0.05). Conclusion:HBx may promote podocyte pyroptosis of HBV-GN via downregulating miRNA-223 targeting NLRP3 inflammasome, suggesting that miRNA-223 is expected to be a potential target for the treatment of HBV-GN.
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Objective:To evaluate the role of silent information regulator-1 (SIRT1)/nucleotide-binding domain (NOD)-like receptor protein-3 (NLRP3) signaling pathway in sevoflurane postconditioning-induced attenuation of oxygen-glucose deprivation and restoration (OGD/R) injury in mouse hippocampal neuronal cell line (HT22) cells.Methods:The HT22 cells were seeded in a culture plate (96-well plate, 100 μl/well; 6-well plate, 2 ml/well) at the density of 5×10 4 cells/ml or in a culture dish (6 cm in diameter) and then divided into 4 groups ( n=24 each) using a random number table method: control group (Control group), OGD/R group, sevoflurane postconditioning group (SPC group), and SIRT1 small interfering RNA group (si-SIRT 1 group). In Control group, cells were cultured at 37 ℃ in normal culture atmosphere. In OGD/R group, the culture medium was replaced with glucose-free serum-free culture medium, and cells were exposed to 95% N 2+ 5% CO 2 for 4 h in an incubator at 37 ℃, and then the glucose-free serum-free culture medium was replaced with the primary culture medium, and cells were cultured for 24 h at 37 ℃ in normal culture atmosphere. In SPC group, the glucose-free serum-free culture medium was replaced with the primary cell culture medium after 4-h oxygen and glucose deprivation, the cells were put into the hypoxia incubator chamber which was filled with 2% sevoflurane immediately after start of reoxygenation, then the chamber was placed in an incubator and the cells were cultured for 1 h at 37 ℃ in normal culture atmosphere, and finally the cells were removed from the chamber and cultured for 23 h at 37 ℃ in normal culture atmosphere. In si-SIRT1 group, SIRT1 small interfering RNA 150 pmol was added at 24 h before surgery, cells were then incubated, and the other procedures were the same as those previously described in group SPC. The cell survival rate was determined using MTT assay. TUNEL assay was used to detect cell apoptosis, and the apoptosis rate was calculated. The expression of SIRT1, NLRP3, IL-1β and IL-18 mRNA was determined using polymerase chain reaction. The expression of SIRT1, NLRP3, interleukin-1beta (IL-1β) and IL-18 was detected using Western blot. Results:Compared with Control group, the cell survival rate was significantly decreased, the apoptosis rate was increased, the expression of SIRT1 protein and mRNA was down-regulated, and the expression of NLRP3, IL-1β and IL-18 protein and mRNA was up-regulated in OGD/R group ( P<0.05). Compared with OGD/R group, the cell survival rate was significantly increased, the apoptosis rate was decreased, the expression of SIRT1 protein and mRNA was up-regulated, and the expression of NLRP3, IL-1β and IL-18 protein and mRNA was down-regulated in SPC group ( P<0.05). Compared with SPC group, the cell survival rate was significantly decreased, the apoptosis rate was increased, the expression of SIRT1 protein and mRNA was down-regulated, and the expression of NLRP3, IL-1β and IL-18 protein and mRNA was up-regulated in si-SIRT1 group ( P<0.05). Conclusions:Activation of SIRT1-NLRP3 signaling pathway is involved in sevoflurane postconditioning-induced attenuation of OGD/R injury in HT22 cells.
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Objective:To evaluate the role of bilateral superior cervical sympathetic ganglia (SCG) in myocardial ischemia-reperfusion (I/R) injury in mice and the relationship with NOD-like receptor protein 3 (NLRP3) inflammasomes.Methods:Thirty-two healthy SPF male C57BL mice, aged 8-10 weeks, weighing 25-30 g, were divided into 4 groups ( n=8 each) by the random number table method: sham operation group (NS group), myocardial I/R group (NIR group), bilateral SCG excision group (SCGx group) and bilateral SCG excision + myocardial I/R group (SCGx+ IR group). The myocardial I/R injury model was prepared by ligating the anterior descending branch of the left coronary artery for 30 min followed by 24 h reperfusion in isoflurane-anesthetized mice. Bilateral superior cervical sympathectomy was performed at 3 days before reperfusion. Blood samples were collected from the inferior vena cava at 24 h of reperfusion for examination of pathological changes (by HE and WGA staining) and for measurement of serum creatine kinase isoenzymes (CK-MB) activity, cardiac troponin I (cTnI) concentration, norepinephrine (NE) concentration and lactic dehydrogenase (LDH) activity (by enzyme-linked immunosorbent assay), superoxide dismutase (SOD) activity (by colorimetric method), myocardial reactive oxygen species (ROS) level (by DHE method), myocardial infarct size(by TTC method), and expression of interleukin-1beta (IL-1β), IL-6, IL-10, tumor necrosis factor-alpha (TNF-α), NLRP3 mRNA (by quantitativepolymerase chain reaction ), and expression of tyrosine hydroxylase (TH), IL-1β, TNF-α, NLRP3, atrial natriuretic peptide (ANP)and brain natriuretic peptide (BNP) (by Western blot). Results:Compared with NS group, the NE concentration was significantly decreased, and TH expression was down-regulated in SCGx group, and the serum CK-MB activity, concentrations of cTnI and NE, LDH activity and myocardial ROS level were significantly increased, SOD activity was decreased, the expression of IL-1β, TNF-α, NLRP3, ANP and BNP was up-regulated, and the expression of IL-1β, IL-6, TNF-α and NLPR3 mRNA was up-regulated in NIR group ( P<0.05). Compared with SCGx group, the serum CK-MB activity, concentrations of cTnI and NE, LDH activity and myocardial ROS levels were significamtly increased, SOD activity was decreased, the expression of IL-1β, TNF-α, NLRP3, ANP and BNP was up-regulated, and the expression of IL-1β, IL-6, TNF-α and NLPR3 mRNA was up-regulated in SCGx+ NIR group ( P<0.05). Compared with NIR group, the serum CK-MB activity, cTnI concentration, LDH activity and myocardial ROS level were significantly decreased, SOD activity was increased, the expression of IL-1β, TNF-α, NLRP3, ANP and BNP was down-regulated, the expression of IL-1β, IL-6, TNF-α and NLPR3 mRNA was down-regulated, and myocardial infarct size was decreased in SCGx+ NIR group ( P<0.05). Conclusions:The mechanism by which bilateral SCG excision attenuates myocardial I/R injury is associated with decreased NLRP3 inflammatory inflammasome activation and inhibition of inflammatory responses in mice.
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Objective:To evaluate the effect of superior cervical ganglion block (SCGB) on cardiac function and nucleotide like receptor protein 3 (NLRP3) signaling pathway in a rat model of myocardial ischemia-reperfusion (I/R).Methods:Sixty healthy SPF male Sprague-Dawley rats, weighing 250-300 g, aged 2-3 months, were divided into 4 groups ( n=15 each) using a random number table method: sham operation group (sham group), myocardial I/R group (IR group), myocardial I/R + normal saline group (IR+ NS group), and myocardial I/R + SCGB group (IR+ SCGB group). Myocardial I/R model was developed by ligation of the left anterior descending branch of the coronary artery for 45 min followed by restoration of blood flow in anesthetized aninals. IR+ SCGB group received SCGB (0.25% ropivacaine 0.1 ml) at 10 min before reperfusion once a day for 2 consecutive weeks, while 0.9% sodium chloride was given instead of ropivacaine in IR+ NS group. Blood samples were collected at 24 h and 14 days of reperfusion for determination of serum concentrations of norepinephrine (NE), troponin T (TnT), tumor necrosis factor-alpha (TNF-α), interleukin-18 (IL-18) and IL-1β by enzyme-linked immunosorbent assay. Echocardiography was performed before ischemia and at 14 days of reperfusion, and left ventricular short axis shortening rate (FS), ejection fraction (EF), and cardiac output (CO) were measured. The rats were sacrificed at 14 days of reperfusion and the hearts were taken for determination of the contents of norepinephrine (NE) in myocardial tissues in the infarction area (by enzyme-linked immunosorbent assay), percentage of myocardial fibrosis area (by Masson staining), M1 macrophage marker CD68 + cell count in the infarction area (by immunohistochemical method), and expression of NLRP3 and gasdermin D (GSDMD) in myocardial tissues (by Western blot). Results:Compared with Sham group, the serum concentrations of TnT, TNF-α, IL-18 and IL-1β, percentage of myocardial fibrosis area, and NE levels in serum and myocardial tissues were significantly increased, the expression of NLRP3 and GSDMD in myocardial tissues was up-regulated, CD68 + cell count was increased, and EF, CO and FS were decreased in IR group ( P<0.05). Compared with IR group, the serum concentrations of TnT, TNF-α, IL-18 and IL-1β, percentage of myocardial fibrosis area, and NE levels in serum and myocardial tissues were significantly decreased, the expression of NLRP3 and GSDMD in myocardial tissues was down-regulated, CD68 + cell count was decreased, and EF, CO and FS were increased in IR+ SCGB group ( P<0.05), and no statistically significant changes were found in the parameters mentioned above in IR+ NS group ( P>0.05). Conclusions:SCGB can improve the cardiac function in a rat model of myocardial I/R, and the mechanism may be related to the inhibition of NLRP3 signaling pathway.
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Objective:To evaluate the role of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) in sepsis-associated encephalopathy (SAE) and the relationship with pyroptosis in microglia of mice.Methods:Twenty-four SPF healthy male C57BL/6J mice, aged 6-8 weeks, weighing 18-22 g, were divided into 3 groups ( n=6 each) using a random number table method: sham operation group (Sham group), SAE group and SAE plus an NLRP3 inhibitor MCC950 group (SAE+ MCC950 group). The mouse model of SAE was prepared by cecal ligation and puncture after anesthesia. MCC950 20 mg/kg was intraperitoneally injected at 1 h after developing the model in SAE+ MCC950 group, and the equal volume of normal saline was given instead in the other groups. Open field tests were conducted at 1 day after developing the model to record the number of rearing and time spent in the central area. Novel object recognition tests were conducted at 2-3 days after developing the model to record the recognition index. After the behavioral experiment on 3 day after developing the model, mice were sacrificed and hippocampal tissues were collected for determination of the expression of NLRP3 (by Western blot), count of cells co-expressing NLRP3 and microglia-specific ionized calcium-binding adaptor molecule 1 (Iba-1) (by immunofluorescence), activity of caspase-1, and contents of interleukin-1beta(IL-1β) and IL-18 (by enzyme-linked immunosorbent assay). Results:Compared with Sham group, the number of rearing was significantly reduced, the time spent in the central area was shortened, the recognition index was decreased, the expression of NLRP3 was up-regulated, the count of NLRP3 + -Iba-1 + cells was increased, and the activity of caspase-1 and contents of IL-1β and IL-18 were increased in SAE and SAE+ MCC950 groups ( P<0.05). Compared with SAE group, the number of rearing was significantly increased, the time spent in the central area was prolonged, the recognition index was increased, the expression of NLRP3 was down-regulated, the count of NLRP3 + -Iba-1 + cells was decreased, and the activity of caspase-1 and contents of IL-1β and IL-18 were decreased in SAE+ MCC950 group ( P<0.05). Conclusions:NLRP3 is involved in the development of SAE, which may be related to the mediation in microglial pyroptosis in mice.
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Objective:To evaluate the relationship between cannabinoid receptor 1 (CB1R) and the NOD-like receptor thermal protein domain-associated protein 3 (NLRP3) during the reduction of cerebral ischemia-reperfusion (I/R) injury by electroacupuncture (EA) preconditioning in rats.Methods:Forty SPF healthy male Sprague-Dawley rats, aged 7-9 weeks, weighing 250-280 g, were divided into 5 groups ( n=8 each) according to the random number table method: sham operation group (Sham group), cerebral I/R group (I/R group), EA preconditioning group (EA group), CB1R antagonist AM251+ EA preconditioning group (AM251+ EA group), and CB1R agonist WIN 55, 212-2 group (WIN group). Cerebral I/R was induced by middle cerebral artery occlusion (MCAO) in anesthetized animals. In EA group, EA preconditioning was performed, and the acupoint Baihui (GV20) was stimulated for 30 min with disperse-dense waves, the intensity of 1 mA and frequency of 2/15 Hz once a day for 5 consecutive days, and the model of cerebral I/R injury was developed at 24 h after the last EA. In AM251+ EA group, CB1R antagonist AM251 1 mg/kg was intraperitoneally injected at 30 min before each stimulation, and the remaining operations were the same as those previously described in EA group. CB1R agonist WIN 55, 212-2 1.5 mg/kg was intraperitoneally injected for 5 consecutive days, and the model of cerebral I/R injury was prepared at 24 h after the last injection in WIN group. Neurological behavior was assessed and scored at 3 days of reperfusion. Then the rats were sacrificed, and brains were removed, and the infarct volume was measured by TTC staining, and the tissues in the ischemic penumbra were extracted for determination of the expression of NLRP3, caspase-1 and interleukin-1bata (IL-1β) by Western blot. Results:Compared with Sham group, the percentage of cerebral infarct volume was significantly increased, the neurobehavioral score was decreased, and the expression of NLRP3, caspase-1 and IL-1β was up-regulated in I/R group ( P<0.05). Compared with I/R group, the percentage of cerebral infarct volume was significantly decreased, the neurobehavioral score was increased, and the expression of NLRP3, caspase-1 and IL-1β was down-regulated in EA and WIN groups ( P<0.05). Compared with EA group, the percentage of cerebral infarct volume was significantly increased, the neurobehavioral score was decreased, and the expression of NLRP3, caspase-1 and IL-1β was up-regulated in AM251+ EA group ( P<0.05). Conclusions:EA preconditioning may inhibit the activation of NLRP3 inflammasomes by activating CB1R, thus alleviating cerebral I/R injury in rats.
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Objective:To evaluate the relationship between Sestrin2 and mitochondrial DNA (mtDNA)-NOD-like receptor associated protein 3 (NLRP3) inflammasome pathway during endotoxin-induced myocardial injury in mice.Methods:One hundred and eighty-four clean-grade healthy male ICR mice, aged 8-12 weeks, weighing 20-25 g, were used in this study. One hundred and sixty-eight mice were divided into 7 groups ( n=24 each) using the random number table method: normal control group (N group), lipopolysaccaride(LPS) group (L group), mtDNA group, LPS+ mtDNA group (M group), normal control+ negative control adeno-associated virus (AAV-NC)group (NC group), LPS+ mtDNA+ AAV-NC group (MC group), and LPS+ mtDNA+ Sestrin2 overexpression adeno-associated virus (AAV-Sestrin2) group (MSgroup). Another 10 mice were used to detect the transfection effect of AAV-Sestrin2, and the left 6 mice were used for mtDNA extraction. The model of endotoxemia was developed by intraperitoneal injection of LPS 10 mg/kg. mtDNA 5 mg/kg was intraperitoneally injected in mtDNA group, and mtDNA 5 mg/kg was intraperitoneally injected at 30 min after LPS injection in M group.AAV-Sestrin2 150 μl was injected via the tail vein in MS group, and the equal volume of AAV-NC was injected via the tail vein in MC and NC groups. Four weeks after virus injection, LPS 10 mg/kg was intraperitoneally injected and 30 min later mtDNA 5 mg/kg was intraperitoneally injected in MS and MC groups. Blood samples were collected at 24 h after LPS injection for determination of serum creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) activities (by biochemical assay), concentrations of serum cardiac troponin I (cTnI), interleukin-18 (IL-18) and interleukin-1beta (IL-1β)(by enzyme-linked immunesorbent assay), and expression of mtDNA (by quantitative real-time polymerase chain reaction). The animals were sacrificed after the end of blood sampling and myocardial tissues were obtained for determination of the contents of reactive oxygen species (ROS), total antioxidant capacity (T-AOC), and adenosine triphosphate (ATP) and expression of NOD-like receptor associated protein 3 (NLRP3), active subunit p20 of caspase-1 (caspase-1p20) and apoptosis-associated microprotein (ASC) in myocardial tissues (by Western blot) and for microscopic examination of the pathological changes after HE staining (with a light microscope). Results:Compared with N group, the levels of CK-MB, LDH, cTnI, IL-1β and IL-18 in serum were significantly increased, the expression of mtDNA was up-regulated, the ROS content in myocardial tissues was increased, the T-AOC and ATP contents in myocardial tissues were decreased, the expression of NLRP3, caspase-1p20 and ASC in the myocardial tissues was up-regulated( P<0.05), and the pathological changes of myocardial tissues were aggravated in L group and mtDNA group.Compared with L group and mtDNA group, the levels of CK-MB, LDH, cTnI, IL-1β and IL-18 in serum were significantly increased, the expression of mtDNA was up-regulated, the ROS content in myocardial tissues was increased, the T-AOC and ATP contents in myocardial tissues were decreased, the expression of NLRP3, caspase-1p20 and ASC in the myocardial tissues was up-regulated( P<0.05), and the pathological changes of myocardial tissues were aggravated in M group. Compared with M group, the levels of CK-MB, LDH, cTnI, IL-1β and IL-18 in serum were significantly decreased, the expression of mtDNA was down-regulated, the ROS content in myocardial tissues was decreased, the T-AOC and ATP contents in myocardial tissues were increased, the expression of NLRP3, caspase-1p20 and ASC in the myocardial tissues was down-regulated( P<0.05), and the pathological changes of myocardial tissues were significantly attenuated in MS group. Conclusions:Sestrin2 can reduce endotoxin-induced myocardial injury in mice by alleviating mitochondrial damage, inhibiting oxidative stress, protecting mtDNA from oxidative damage, and then inhibiting mtDNA-NLRP3 inflammasome pathway.
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Objective: To explore the influence factors of poor prognosis of esophageal squamous cell carcinoma (ESCC) and the predictive value of inflammatory reaction indexes including neutrophils and lymphocytes ratio (NLR), platelet and lymphocyte ratio (PLR), monocyte and lymphocyte ratio (MLR) provision and differentiation degree, infiltration depth, lymph node metastasis number on the postoperative recurrence of ESCC. Methods: A total of 130 patients with ESCC who underwent radical resection from February 2017 to February 2019 in Nanyang Central Hospital were selected and divided into good prognosis group (66 cases) and poor prognosis group (64 cases) according to the prognostic effect. The clinical data and follow-up data were collected. Multivariate logistic regression analysis was used to determine the independent influencing factors of poor prognosis. Spearman correlation analysis was used to determine the correlation between preoperative NLR, PLR and MLR with the degree of differentiation, depth of invasion and number of lymph node metastases. Receiver operating characteristic (ROC) curve analysis was used to evaluate the efficacy of NLR, PLR and MLR in predicting poor prognosis of ESCC. Results: Univariate analysis showed that the degree of differentiation, the degree of invasion and the number of lymph node metastasis were related to the prognoses of patients with ESCC (P<0.05). Multivariate logistic regression analysis showed that the degree of differentiation, depth of invasion and number of lymph node metastases were independent influencing factors for poor prognosis of patients with ESCC, moderate differentiation (OR=2.603, 95% CI: 1.009-6.715) or low differentiation (OR=9.909, 95% CI: 3.097-31.706), infiltrating into fibrous membrane (OR=14.331, 95% CI: 1.333-154.104) or surrounding tissue (OR=23.368, 95% CI: 1.466-372.578), the number of lymph node metastases ≥ 3 (OR=9.225, 95% CI: 1.693-50.263) indicated poor prognosis. Spearman correlation analysis showed that NLR was negatively correlated with the degree of differentiation and the number of lymph node metastases (r=-0.281, P=0.001; r=-0.257, P=0.003), PLR was negatively correlated with the degree of differentiation, depth of invasion and number of lymph node metastasis (r=-0.250, P=0.004; r=0.197, P=0.025; r=-0.194, P=0.027), MLR was positively correlated with the degree of differentiation and the number of lymph node metastasis (r=0.248, P=0.004; r=0.196, P=0.025). ROC curve analysis showed that the areas under the curve of NLR, PLR and MLR in predicting poor prognosis of ESCC were 0.971, 0.925 and 0.834, respectively. The best cut-off value of NLR was 2.87. The sensitivity and specificity of NLR in predicting poor prognosis of ESCC were 90.6% and 87.9%, respectively. The optimal cut-off value of PLR was 141.75. The sensitivity and specificity for predicting poor prognosis of ESCC were 92.2% and 87.9%, respectively. The best cut-off value of MLR was 0.40. The sensitivity and specificity of MLR in predicting poor prognosis of esophageal squamous cell carcinoma were 54.7% and 100.0%, respectively. Conclusions: The degree of differentiation, the degree of invasion and the number of lymph node metastases are closely related to the poor prognosis of patients with esophageal squamous cell carcinoma. NLR, PLR and MLR can provide important information for predicting the poor prognosis of esophageal squamous cell carcinoma.
Subject(s)
Humans , Esophageal Squamous Cell Carcinoma/pathology , Prognosis , Lymphatic Metastasis/pathology , Esophageal Neoplasms/pathology , Neutrophils , Lymphocytes , Blood Platelets/pathology , Inflammation , Retrospective StudiesABSTRACT
Background:Aim of the study was to analyse the relation between elevated neutrophil lymphocyte ratio (NLR), and erythrocyte sedimentation rate (ESR) in Helicobacter pylori(H. pylori)positive chronic gastritis patients, as compared to the control group containing H. pylorinegative chronic gastritis patients.Methods:Chronic gastritis patients were segregated in equal numbers based on H. pyloristatus. NLR was calculated, and ESR noted from the observations, comparison was done between the control and the study groups.Results:A total of 100 patients were included in the study. The 50 each from the control and study group. An observation of elevation in NLR and ESR in H. pyloripositive chronic gastritis patients, as compared to the control group was seen. With an average NLR of 2.43 and 1.43, in the control and study group, respectively.Conclusions:Raise in NLR in H. pyloripositive chronic gastritis patients with an associated raise in ESR suggests, the severity of the infection and the need for eradication and prevent complications
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Background:Lung cancer is ranked third as the most common cancer in Indonesia. The one-year survival rate of advanced-stage non-smallcell lung carcinoma (NSCLC) patient is quite low, that is 24.6%. Effective and inexpensive prognostic markers need to be further studied due to an increasing incidence of cancer. Inflammation plays an important role in tumorigenesis and research showed an association of NLR and PLR values with poor prognosis in patients with various solid tumors, but current cutoff values still vary. This study wanted to determine the value of NLR, PLR and their relationship with the survival rate of advancedstage NSCLC patients at Sanglah hospital.Methods:A retrospective cohort study using the medical record of 96 advanced-stage NSCLC patients who underwent treatment since January 2018 in Sanglah hospital, was closely monitored for a year since diagnosed. Analysis was performed with ROC, Kaplan Meier analysis, log rank test, and time independent cox regression model.Results:The one-yearsurvival rate of advancedstage NSCLC patient is 14.6%, with median survival 3.26 months. Cut off NLR> 3.37, median survival 2.66,p=0.00. Cut off PLR>178.55, median survival 3.26, p=0.35. Multivariate analysis showed that NLR, HR=2.75 and performance status, HR=1.78 were associated with survival.Conclusions:NLR with cut off>3.37 is associated with one-year survival rate of advanced-stage NSCLC patient. PLR did not have any significant association with one-year survival rate of advanced-stage NSCLC patient.
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Background:The objective of this study was to assess the demographical characteristic, laboratory and radiological findings associated with COVID-19 mortality in hospitalizedpatients and also to co-relate neutrophil-to-lymphocyte ratio (NLR)and chest x-ray (CXR)score with severity of the disease.Methods:This is a retrospective study done in Bowring and Lady Curzon hospital between the periodof May 2021 to July 2021. 100 patients who were tested positive for SARS-CoV2 with RT-PCR were taken for the study after fulfilling the inclusion criteria. On day 1 of admission, routine blood investigations including CBC with differential count and chestX ray is taken. From the above said data, NLR and CXR score is calculated and a comparison is made to determine severity and in-hospital mortality between mild, moderate and severe COVID pneumonia patients. This study is being carried out after obtaining institutional ethical committee approval clearance. All analysis were performed using SPSS software version 10.Results: The sample size studied was 100. The mean age of patients was 28.3 in mild, 49.9 in moderate and 62.6 in severe COVID patients. Among these 67% were males and 33% were females. It was noted that, leukocytosis(mean-13245), neutrophilia (mean-83.05%), lymphocytopenia (mean-10.45%) and chest X-ray score (mean-4.98) was seen among severe group with p value being significant.Conclusions: TLC, NLR and CXR score were significantly different between severe and non-severe patients, so assessment of these simple parameters may help identify high risk COVID-19 patients at an early stage in a resource limited setting from the data retrieved from our hospital, NLR and CXR Score showed an acceptable efficiency to separate COVID-19 patients among severe and non-severe patients with a significant p value thereby helping in triaging the patients and need for early ICU needs.
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Neutrophil-lymphocyte ratio (NLR) has been a novel biomarker in the literature over decades to assess systemic inflammation. This easily available and cost-effective marker has shown clinical significance in various disease conditions such as infections, sepsis, malignancies, cardiovascular events and even in SARS-COVID-19 pneumonia. NLR has a major association in ischemic stroke patients. Inflammation is a crucial step in the development of ischemic stroke, with neutrophils and lymphocytes comprising the first line defence mechanism. It has a role in stroke initiation, progression of injury, and recovery. This article showed significant correlation between NLR and stroke patients in terms of days of hospital stay, severity and prognosis. High NLR values were found to be associated with higher risk of stroke, prolonged hospital stay and a worse 3 month mortality rate. NLR can thus be integrated into any clinical practice, having an impact on early diagnosis in many clinical scenarios.
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Background:COVID-19 pandemic has strained the health infrastructure globally, hence the importance of cost-effective biomarkers. We aimed to identify simple haematological prognostic markers in hospitalized COVID-19 patients to differentiate between milder and severe cases, thus predicting outcome.Methods:A retrospective study of COVID-19 patients admitted at MallaReddy institute of medical sciences was conducted from April to June 2021. Total leukocyte count (TLC), neutrophil-to-lymphocyte ratio (NLR), derived NLR ratio (dNLR) and platelet-to-lymphocyte ratio (PLR) were calculated and correlated with outcome. These parameters were compared with other inflammatory markers using ROC(receiver operator curve)analysis.Results:303 patients of 397 fulfilled the inclusion criteria (male-198, female-105). There was a significant higher mean of NLR in patients with death (14.46�84) compared to patients recovered (8.43�33), similarly the dNLR was higher in death (8.06�34) compared to recovered (4.97�49). A significant positive strength of association between the NLR and dNLR with the ESR, CRP, CORADS score and CT severity score in the patients. The ROC analysis showed the NLR (AUC=0.777) and dNLR (0.799) a better marker to predict the outcome.Conclusions:In COVID-19, immuno-haematological markers like NLR, dNLR, PLR found to be a simple and cost-effective tool to prognosticate the clinical outcome among hospitalized patients and were in concordance with the other inflammatory markers. Hence, these markers serve as better indicators in risk stratification and better management.
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ABSTRACT BACKGROUND: Quick and accurate identification of critically ill patients ensures appropriate and correct use of medical resources. In situations that threaten public health, like pandemics, rapid and effective methods are needed for early disease detection among critically ill patients. OBJECTIVE: To determine the relationship between the neutrophil-to-lymphocyte ratio (NLR) of coronavirus disease-19 (COVID-19) patients upon admission to the emergency department (ED) and these patients' prognosis. DESIGN AND SETTING: Retrospective cohort study among COVID-19 patients in the ED of a tertiary-level hospital. METHODS: Data on patients' age, gender, vital signs, chronic diseases, laboratory tests and clinical outcomes were collected from electronic medical records. Receiver operating characteristic (ROC) curve analysis was performed. The area under the curve (AUC) was used to assess the accuracy of NLR for predicting in-hospital mortality risk and intensive care unit (ICU) requirement. The Youden J index (YJI) was used to determine optimal threshold values. RESULTS: 1,175 patients were included. Their median age was 63 years (IQR, 48-75). With an NLR cutoff value of 5.14, the sensitivity, specificity, PPV, AUC and YJI for ICU requirement were calculated as 77.87%, 74.08%, 92.4%, 0.811 and 0.5194, respectively. With the same cutoff value, the sensitivity, specificity, AUC and YJI for in-hospital mortality were 77.27%, 75.82%, 0.815 and 0.5309, respectively. In addition, advanced age, leukocytosis, anemia and lymphopenia were found to be associated with poor prognosis. CONCLUSION: The NLR, which is a widely available simple parameter, can provide rapid insights regarding early recognition of critical illness and prognosis among COVID-19 patients.
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Humans , COVID-19 , Prognosis , Lymphocytes , Retrospective Studies , ROC Curve , SARS-CoV-2 , Middle Aged , NeutrophilsABSTRACT
【Objective】 To study the value of neutrophil/lymphocyte ratio (NLR) and platelet/lymphocyte ratio (PLR) in the diagnosis and prediction of disease severity in adolescent atopic dermatitis (AD). 【Methods】 A total of 100 AD patients and 50 healthy individuals were selected from Dermatology Department of our hospital.The age, gender, blood routine results of all subjects were collected, and NLR and PLR were calculated.The diagnostic value of NLR and PLR for AD and severe AD was discussed by receiver operating curve (ROC), the relationship between NLR, PLR and the risk of AD was analyzed. 【Results】 The AUC of NLR and PLR in the diagnosis of AD was 0.780 and 0.769, with the best cutoff value at 2.95 and 98.50 respectively, and no significant difference in the predictive value between NLR and PLR was noticed (P>0.05). The proportion of NLR≥2.95, PLR≥98.50, red blood cell distribution width (RDW), mean platelet volume (MPV), white blood cell count (WBC) and eosinophils (E) in AD group were significantly higher than those in control group (all P<0.05). Before and after adjusting for confounding factors, the odds ratio (OR) of patients with NLR≥2.95 and PLR≥98.50 were 12.250 vs 6.048 and 5.525 vs 4.352, respectively.NLR, PLR and E in mild AD group were lower than those in severe AD group, and NLR and E in moderate AD group were lower than those in severe AD group, the differences were statistically significant (all P<0.05). The AUC of NLR and PLR in diagnosis of severe AD was respectively 0.712 and 0.675, the difference was statistically significant (P<0.05). NLR, PLR, RDW and E were positively correlated with SCORAD score, and the r values were 0.254, 0.198, 0.202 and 0.219 respectively. 【Conclusion】 The increase of NLR and PLR were the risk factors of AD, and had certain diagnostic value for AD and severe AD.