Objective To investigate the expression of FLIP in the lung of rats and the protective effect in development of acute lung injury( ALI) with the adenovirus vector carrying FLIP gene( Ad-FLIP)inhaled. Methods Forty-eight rats were randomly divided into four groups, with 12 rats in each gruop. In treatment group, ALI rats model was eatablished by LPS intraperitoneal injection and then inhaled Ad-FLIP vector. In prevention group, the animals were infected with Ad-FLIP vector before ALI model wasestablished. Two control groups of treatment and prevention received Ad-EGFP vectors respectively.Pathological changes of lung were observed under light microscope. Wet/dry weight ( W/D) of lung lobes and lung permeability index( LPI) were also measured. The mRNA and protein expressions of FLIP in lungwere investigated by RT-PCR and immunohistochemistry, respectively. Results Lung histopathological changes were alleviated, the index of W/D and LPI were significantly lower, the expressions of FILP mRNA and protein in the lung were elevated both in the treatment group and prevention group compared to thecontrol groups ( all P lt;0. 01) . Conclusion Ad-FLIP transfection can up-regulate the expression of FLIP in lung of rats, and might protect respiratory membrane and lessen pulmonary edema to prevent the development of ALI.
Diabetes has become a global epidemic disease now. Its chronic progressive deterioration and the acute and chronic complications affect the quality of the patients' lives seriously. The prevention and treatment of diabetes has become one of the research focuses in recent years. NLRP3 (NOD-like receptor family, pyrin domain containing 3) inflammasome can recognize the metabolic stress signals, and cause caspase-1 activation and interleukin-1β (IL-1β) production, and is closely related to diabetes development. The latest studies have shown that NLRP3 inflammasome will be a new potential target for the treatment of diabetes. This article reviews the activation and regulation of NLRP3 inflammasome, and the effect of NLRP3 inflammasome on glucose metabolism and its targeted therapy in diabetes.
Objective To investigate interleukin-1β (IL-1β), IL-6, and IL-17 levels in both synovial fluid and serum of patients with primary knee medial osteoarthritis (OA) after high tbial osteotomy (HTO). Methods Twenty-six patients with primary knee medial OA undergoing HTO between January 2011 and June 2014 (experimental group) and 30 healthy individuals (control group) were recruited into the study. There was no significant difference in gender, age, and body mass index between 2 groups (P>0.05). The X-ray film was taken to record healing time at osteotomy site, to measure the tibiofemoral angle, and to assess limb alignment after HTO. Visual analogue scale (VAS) pain score and knee society score (KSS) were used to evaluate pain level and function of the knee. The IL-1β, IL-6, and IL-17 concentrations in both plasma and synovial fluid were measured before operation and at 6, 12, and 18 months after operation in the experimental group using ELISA method; the levels in plasma were measured in control group. Results Primary healing of incisions was achieved in patients. All patients were followed up 18-24 months (mean, 21 months). The X-ray film showed osseous healing at osteotomy site at 9-14 weeks (mean, 11.5 weeks). The average tibiofemoral angle was 167.5° (range, 165-170°) after bone healing. Satisfactory limb alignment was obtained in all patients. The postoperative VAS pain score was significantly decreased and KSS score was significantly improved when compared with preoperative scores (P<0.05), but no significant difference was found between different time points after operation (P>0.05). The preoperative plasma and synovial fluid IL-1β, IL-6, and IL-17 concentrations were significantly higher in patients than controls (P<0.05). The postoperative IL-1β, IL-6, and IL-17 concentrations in plasma and synovial fluid were significantly lower than preoperative ones in patients (P<0.05), but the concentrations were significantly higher than those in controls (P<0.05). The postoperative plasma and synovial fluid IL-1β, IL-6, and IL-17 concentrations were significantly declined in patients, but there was no significant difference between different time points after operation (P>0.05). Conclusion HTO can significantly improve the pain symptom and joint function and reduce IL-1β, IL-6, and IL-17 levels in both plasma and synovial fluid of patients with medial compartment knee OA, but these cytokines can not return to normal level.
The aim of this article is to study how andrographolide-releasing collagen scaffolds influence rabbit articular chondrocytes in maintaining their specific phenotype under inflammatory environment. Physical blending combined with vacuum freeze-drying method was utilized to prepare the andrographolide-releasing collagen scaffold. The characteristics of scaffold including its surface morphology and porosity were detected with environmental scanning electron microscope (ESEM) and a density instrument. Then, the release of andrographolide from prepared scaffolds was measured by UV-visible spectroscopy. Rabbit chondrocytes were isolated and cultured in vitro and seeded on andrographolide-releasing collagen scaffolds. Following culture with normal medium for 3 d, seeded chondrocytes were cultured with medium containing interleukin-1 beta (IL-1β) to stimulate inflammation in vitro for 7 d. The proliferation, morphology and gene transcription of tested chondrocytes were detected with Alamar Blue assay, fluorescein diacetate (FDA) staining and reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR) test respectively. The results showed that the collagen scaffolds prepared by vacuum freeze-dry possess a high porosity close to 96%, and well-interconnected chambers around (120.7±17.8) μm. The andrographolide-releasing collagen scaffold continuously released andrographolide to the PBS solution within 15 d, and collagen scaffolds containing 2.22% andrographolide significantly inhibit the proliferation of chondrocytes. Compared with collagen scaffolds, 0.44% andrographolide-containing collagen scaffolds facilitate chondrocytes to keep specific normal morphologies following 7 d IL-1β induction. The results obtained by RT-qPCR confirmed this effect by enhancing the transcription of tissue inhibitor of metalloproteinase-1 (TIMP-1), collagen II (COL II), aggrecan (Aggrecan) and the ratio of COL II/ collagen I(COL I), meanwhile, reversing the promoted transcription of matrix metalloproteinase-1 (MMP-1) and matrix metalloproteinase-13 (MMP-13). In conclusion, our research reveals that andrographolide-releasing (0.44%) collagen scaffolds enhance the ability of chondrocytes to maintain their specific morphologies by up-regulating the transcription of genes like COL II, Aggrecan and TIMP-1, while down-regulating the transcription of genes like MMP-1 and MMP-13 which are bad for phenotypic maintenance under IL-1β simulated inflammatory environment. These results implied the potential use of andrographolide-releasing collagen scaffold in osteoarthritic cartilage repair.
ObjectiveTo summarize the research advances of pyroptosis in hepatic ischamia-reperfusion injury (IRI).MethodThe literatures about the studies of mechanism of pyroptosis in hepatic IRI were retrieved and analyzed.ResultsPyroptosis, also known as inflammatory necrocytosis, was proven to play an important role in the hepatic IRI. When hepatic ischemia-reperfusion occurred, the classical pathway of pyroptosis dependenting on caspase-1 and the non-classical pathway of pyroptosis dependenting on caspase-11 were initiated by specific stimulants, and leaded to the activation of gasdermin D, releases of proinflammatory factors such as interleukin-1β, interleukin-18, etc., and the recruitment and activation of neutrophils. Consequently, pyroptosis caused more severe hepatic inflammation and aggravated existing cell injury and dysfunction of liver during hepatic IRI.ConclusionsPyroptosis plays an important role in liver IRI. Further researches about mechanism of pyroptosis will be beneficial to the prevention and treatment of the pyroptosis of related diseases.
Objective To explore the effects of low-intensity pulsed ultrasound (LIPUS) on anabolism, apoptosis and intraflagellar transport 88 (IFT88) expression in mouse chondrocytes after interleukin (IL)-1β intervention, and the correlation of cartilage repairment by LIPUS with primary cilia. Methods IL-1β intervention, LIPUS intervention and lentiviral carrying IFT88-specifific short hairpin RNA (sh-IFT88) transfection were performed on mouse chondrocytes, respectively. The groups included: normal chondrocyte group (N group), chondrocyte after IL-1β intervention group (OA group), chondrocyte after IL-1β intervention+LIPUS group (OA+U group), sh-IFT88+IL-1β intervention chondrocyte group (KO+OA group), and sh-IFT88+LIPUS+IL-1β treated chondrocyte group (KO+OA+U group). Real-time polymerase chain reaction and immunofluorescence were used to determine the expression of collagen Ⅱ, aggrecan, and primary cilia, and apoptosis was measured by flow cytometry. All experimental data were statistically analyzed using the GraphPad Prism 9.5 software. Results The expression of collagen Ⅱ and aggrecan increased, the apoptosis decreased, and the incidence of primary cilia in chondrocytes of mice increased in the OA+U group compared with those in the OA group (P<0.05). The collagen Ⅱ and aggrecan expression decreased and the apoptosis increased in the KO+OA+U group compared with those in the OA+U group (P<0.05). Conclusion LIPUS can reduce the apoptosis of chondrocytes in C57 mice after IL-1β intervention, and increase the expression of collagen Ⅱ and aggrecan in chondrocyte matrix, and the effect is related to primary cilia.