摘要
OBJECTIVE To explore the expression changes of TLR4/MyD88/NF-κB signaling pathways in olfactory disorders.METHODS There were 40 healthy BALB/c mice who were divided into an observation group and a control group,with 20 mice in each group.Detection of Toll-like receptors(TLR4),myeloid differentiation primary response gene 88(MyD88)and nuclear factor kappa B(NF-κB)in mice using quantitative reverse transcription PCR level;Detection of TLR4,MyD88 and NF-κB by Western blot(WB)test protein content;Immunohistochemical detection of the expression of mouse olfactory marker protein(OMP).RESULTS There was no significant difference in foraging time between the two groups of mice before modeling(P>0.05),after modeling,the foraging time of the observation group mice was significantly longer than that of the control group(P<0.05);The relative mRNA expression level of TLR4,MyD88 and NF-κB in the nasal epithelium of mice in the observation group was significantly higher than that of the control group(P<0.05);The protein expression of TLR4,MyD88 and NF-κB in the nasal epithelium of mice in the observation group was significantly higher than that of the control group(P<0.05);The level of OMP protein in the nasal epithelium of the observation group was significantly lower than that of the control group(P<0.05).CONCLUSION Expression reinforcement of TLR4/MyD88/NF-κB signaling pathway in a mouse model of olfactory dysfunction.
摘要
ObjectiveTo investigate the therapeutic effect of Qingjie Huagong decoction （QJHGD） on a mouse model of severe acute pancreatitis （SAP） and the mechanism of action of QJHGD against inflammatory response. MethodsA total of 36 male C57BL/6J mice were randomly divided into blank group， model group， Western medicine group （ulinastatin）， and low-， middle-， and high-dose QJHGD groups， with 6 mice in each group. All mice except those in the blank group were given 5% sodium taurocholate by retrograde pancreaticobiliary injection to establish a model of SAP. After modeling， the mice in the low-， middle-， and high-dose groups were given QJHGD （1， 2， and 4 g/kg， respectively） by gavage， and those in the Western medicine group were given intraperitoneal injection of ulinastatin （5×104 U/kg）， for 7 days in total. HE staining was used to observe the histopathological changes of the pancreas； ELISA was used to measure the levels of α-amylase， lipase， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， interleukin-18 （IL-18）， and tumor necrosis factor-α （TNF-α） in mice； RT-qPCR was used to measure the mRNA expression levels of NOD-like receptor protein3 （NLRP3）， Toll-like receptor 4 （TLR4）， and nuclear factor-kappa B （NF-κB） in pancreatic tissue； immunohistochemistry was used to measure the positive expression rates of NLRP3， TLR4， and NF-κB in pancreatic tissue； Western blot was used to measure the protein expression levels of NLRP3， TLR4， NF-κB， IL-1β， and IL-6. An analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the blank group， the model group had diffuse destruction of pancreatic tissue structure， focal dilatation of pancreatic lobular septum， pancreatic acinar atrophy， and massive inflammatory cell infiltration， as well as significant increases in the content of α-amylase， lipase， IL-1β， IL-6， IL-8， IL-18， and TNF-α （all P<0.05）， the mRNA expression levels and positive expression rates of NLRP3， TLR4， and NF-κB （all P<0.05）， and the protein expression levels of NLRP3， TLR4， NF-κB， IL-1β， and IL-6 （all P<0.05）. Compared with the model group， the low-， middle-， and high-dose QJHGD groups and the Western medicine group had slightly tighter and more intact structure of pancreatic tissue， ordered arrangement of pancreatic acinar cells， a small amount of inflammatory cell infiltration， and hemorrhagic foci of pancreatic lobules， as well as significant reductions in the content of α-amylase， lipase， IL-1β， IL-6， IL-8， IL-18， and TNF-α （all P<0.05）， the mRNA expression levels and positive expression rates of NLRP3， TLR4， and NF-κB （all P<0.05）， and the protein expression levels of NLRP3， TLR4， NF-κB， IL-1β， and IL-6 （all P<0.05）. ConclusionQJHGD may exert a protective effect on the pancreatic tissue of SAP mice by inhibiting the activation of NLRP3/TLR4/NF-κB signaling pathway-related proteins， reducing the release of inflammatory mediators， and preventing the enhancement of inflammatory cascade response.
摘要
ObjectiveTo investigate the effect and mechanism of echinacoside （ECH） in improving liver injury in rats with acute pancreatitis by establishing a rat model of acute pancreatitis and liver injury. MethodsA total of 24 Sprague-Dawley rats were randomly divided into blank group （Con group）， control group （Con+ECH group）， acute pancreatitis group （AP group）， and acute pancreatitis+ECH intervention （AP+ECH group）. The rats were given intraperitoneal injection of 10 mg/kg ECH on day 7 before the establishment of the model of acute pancreatitis； at 24 hours after the last administration of cerulein， blood samples were collected via the abdominal aorta， and serum was separated for biochemical analysis including alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， lactate dehydrogenase （LDH）， gamma-glutamyl transpeptidase （GGT）， alkaline phosphatase （ALP）， albumin （Alb）， total bilirubin （TBil）， cholinesterase， blood amylase （Amy）， and lipase （LPS）. HE staining was used to observe the histopathological changes of the pancreas and the liver； transmission electron microscopy （TEM） was used to observe the microstructural changes of pancreas and liver tissue； ELISA was used to measure the levels of interleukin-1β （IL-1β）， interleukin-16 （IL-6）， tumor necrosis factor-α （TNF-α）， and interleukin-10 （IL-10） in liver tissue homogenate； immunohistochemistry was used to measure the levels of TNF-α and p-p65 NF-κB in pancreas and liver tissue； Western blot was used to measure the expression levels of NF-κB pathway proteins in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the SNK test or the Dunnett’s T3 method was used for further comparison between two groups. ResultsCompared with the Con group， the AP group had significant increases in ALT， AST， GGT， LDH， ALP， TBil， Amy， and LPS （all P<0.01）， as well as significant increases in the levels of IL-1β， IL-6， IL-10， and TNF-α in liver tissue homogenate （all P<0.01）. ECH intervention reduced the levels of ALT， AST， GGT， LDH， ALP， TBil， AMY， and LPS and inhibited the secretion of IL-1β， IL-6， and TNF-α in rats with acute pancreatitis. HE staining showed that ECH intervention alleviated the vacuolar degeneration of acinar cells， inflammatory cell infiltration in pancreatic tissue， and the necrosis of hepatocytes compared with the AP group. TEM showed that compared with the AP group， there was a reduction in the degree of mitochondrial swelling in liver and pancreatic cells after ECH intervention. ECH intervention partially reversed the elevated expression levels of p-p65 NF-κB and TNF-α in liver and pancreatic tissue. In addition， the expression levels of MyD88， p-IκBα， p-IKKα， and p-p65 were upregulated in liver tissue of rats with acute pancreatitis， which could be partially reversed after ECH intervention. ConclusionEchinacoside can alleviate liver and pancreatic injury induced by acute pancreatitis by inhibiting the TLR4/MyD88/NF-κB pathway.
摘要
Objective:To explore the effects of apigenin on apoptosis and hypoxia inducible factor-1α (HIF-1α)/nuclear factor κB (NF-κB) signaling pathway in renal cancer A498 cells.Methods:Human renal cell carcinoma A498 cells were cultured in vitro and divided into different concentrations of apigenin (10, 20, 40 μmol/L) groups, apigenin (40 μmol/L)+ HIF-1α agonist dimethylenediaminoacetic acid (DMOG) group, HIF-1α inhibitor rifiximab (YC-1) group, and control group. Cell proliferation was detected using cell counting kit-8 (CCK-8) assay and plate clone formation assay, apoptosis was detected using Hoechst 33258 staining and flow cytometry, and expression of apoptotic proteins and HIF-1/NF-B pathway proteins was detected using Western blot assay.Results:Celery extract significantly inhibited the proliferation of A498 cells, and the inhibitory effect was concentration dependent ( P<0.001). Compared with the control group, the apoptosis rates of A498 cells in the 10, 20, and 40 μmol/L apigenin groups and YC-1 groups were significantly increased [(4.35±1.04)% vs (10.06±1.13)%, (18.52±2.58)%, (32.17±2.63)%, (26.94±2.41)%], as well as the expression levels of B lymphocyte tumor 2 related protein (Bax) and Cleaved Caspase-3 protein, while the expression levels of B lymphocyte tumor 2 (Bcl-2) were significantly reduced (all P<0.001). Compared with the control group, the HIF-1α protein expression levels (0.85±0.08 vs 0.63±0.06, 0.31±0.03, 0.16±0.02) and p-NF-κB p65/NF-κB p65 ratio (0.82±0.08 vs 0.51±0.05, 0.30±0.03, 0.13±0.01) of A498 cells in the 10, 20, and 40 μmol/L apigenin groups were significantly reduced (all P<0.001). Compared with the apigenin group, the apoptosis rate of A498 cells in the apigenin+ DMOG group was significantly reduced [(32.17±2.63)% vs (14.85±1.62)%], and the expression levels of Bax and Cleared Caspase-3 proteins were significantly reduced, while the expression levels of Bcl-2 proteins were significantly increased (all P<0.001). Conclusions:Apigenin may promote apoptosis in renal cancer A498 cells by inhibiting the activation of the HIF-1α/NF-κB signaling pathway.
摘要
Abstract Background: Trimethylamine N-oxide (TMAO), a gut microbiota metabolite, is associated with cardiovascular disease (CVD) development. TMAO can trigger an inflammatory response by inducing the nuclear factor-kappa B (NF-κB) signaling cascade and increasing the expression of pro-inflammatory cytokines, contributing to the worsening of CVD. This study aimed to evaluate the association between TMAO plasma levels and inflammation in patients with coronary artery disease (CAD). Methods: A cross-sectional study was carried out including 29 patients with CAD. Peripheral blood mononuclear cells (PBMC) were isolated from fasting blood samples, and NF-κB and vascular cell adhesion protein 1 (VCAM1) mRNA expression were estimated using real-time quantitative PCR. We determined TMAO plasma levels by LC-MS/MS and TNF-α by ELISA. Routine biochemical parameters were evaluated using an automatic biochemical analyzer. Correlations were estimated by Spearman or Pearson test. Statistical significance was set at the level of p < 0.05. Results: All patients presented TMAO levels within the normal range according to EUTox (normal range: 2.83 ± 1.53 mg/L; CAD patients: 0.2 [0.1 to 0.2] ng/μL). TMAO plasma levels were positively correlated with NF-κB mRNA expression (0.555; p = 0.002). Conclusion: TMAO plasma levels may be associated with NF-κB mRNA expression in patients with CAD and may contribute to the pathogenesis of this disease.
摘要
Abstract Dental pulp stem cells (DPSCs) and periodontal ligament stem cells (PDLSCs) can differentiate into osteoblasts, indicating that both are potential candidates for bone tissue engineering. Osteogenesis is influenced by many environmental factors, one of which is lipopolysaccharide (LPS). LPS-induced NF-κB activity affects the osteogenic potencies of different types of MSCs differently. This study evaluated the effect of LPS-induced NF-κB activity and its inhibition in DPSCs and PDLSCs. DPSCs and PDLSCs were cultured in an osteogenic medium, pretreated with/without NF-κB inhibitor Bay 11-7082, and treated with/without LPS. Alizarin red staining was performed to assess bone nodule formation, which was observed under an inverted light microscope. NF-κB and alkaline phosphatase (ALP) activities were measured to examine the effect of Bay 11-7082 pretreatment and LPS supplementation on osteogenic differentiation of DPSCs and PDLSCs. LPS significantly induced NF-κB activity (p = 0.000) and reduced ALP activity (p = 0.000), which inhibited bone nodule formation in DPSCs and PDLSCs. Bay 11-7082 inhibited LPS-induced NF-κB activity, and partially maintained ALP activity and osteogenic potency of LPS-supplemented DPSCs and PDLSCs. Thus, inhibition of LPS-induced NF-κB activity can maintain the osteogenic potency of DPSCs and PDLSCs.
摘要
Abstract Objective To evaluate whether antimicrobial photodynamic therapy (aPDT) repairs bisphosphonate-related osteonecrosis of the jaw (BRONJ) modulated by the reduction of NF-kB protein in a murine model. Methodology Male Wistar rats (N=30) were divided into the following groups (n=6/group): negative control (NC); experimental osteonecrosis (ONE); ONE + photosensitizer (PS); ONE + photobiomodulation (PBM); and ONE + aPDT. Over 8 weeks, ONE was induced by zoledronic acid 250 µg/kg injections, except in the NC group, which received sterile 0.9% saline, followed by extraction of the lower left first molar. Red light laser irradiation (wavelength ~660 nm, power 50 mW, energy of 2 J, energy dose of 66.67 J/cm2 for 40 s) was performed once a week for 4 weeks. Methylene blue 0.3% was used as PS. The animals were euthanized and examined macroscopically for the presence of exposed bone and epithelial repair and microscopically by histochemical (hematoxylin-eosin and Masson's trichrome staining) and immunohistochemical (anti-NF-kB) methods. Macroscopic and histomorphometric data were analyzed by one-way ANOVA and Tukey's post-test (p<0.05). Results Mucosal repair, viable osteocytes, and NF-kB immunostaining were observed in the NC, ONE+PS, ONE+PBM, and ONE+aPDT groups. The ONE group showed no mucosal repair, showing empty lacunae and multifocal immunostaining for NF-kB. The ONE+PBM and ONE+aPDT groups had greater deposition of extracellular matrix and less necrotic bone tissue (p<0.05). Conclusion PBM and aPDT treatments for BRONJ were effective for bone and epithelial repair, in addition to reducing inflammation mediated by the decrease of NF-kB protein in the irradiated regions.
摘要
El factor nuclear κB (NF-κB) es una familia de factores de transcripción sumamente importantes que regulan una gran variedad de genes en diferentes procesos de las respuestas inmunitarias e inflamatorias. Esta familia está compuesta por cinco miembros relacionados estructuralmente, y pueden inducir la transcripción de genes diana al unirse a segmentos específicos de ácido desoxirribonucleico (ADN). Las proteínas NF-κB son usualmente secuestradas en el citoplasma por una familia de proteínas inhibidoras, sin embargo, diversas vías de señalización oncogénica pueden activarla y desencadenar fenotipos malignos en las células correspondientes. El objetivo principal de esta revisión es comprender los mecanismos de regulación del factor de transducción NF-κB, su patogénesis y sus posibles blancos terapéuticos en cáncer. Se consultaron diferentes bases de datos que incluyeron PubMed, Scopus y SciELO, desde el año 2000 hasta diciembre del año 2022; se buscaron las referencias bibliográficas en relación con las palabras clave asociadas al factor NF-κB y cáncer, para finalmente desarrollar la revisión. El factor nuclear de transcripción NF-κB es importante en muchas vías de señalización celular, participa en diversos procesos biológicos y sus alteraciones están asociadas a trastornos inmunitarios y cáncer, entre otras patologías. NF-κB se expresa en todos los tipos de células y tejidos, de tal forma que muchas mutaciones oncogénicas contribuyen a la activación de NF-κB en las células tumorales, y son nuevas rutas de investigación terapéuticas para el cáncer. Existen dos vías de señalización diferentes de NF-κB, denominadas vía canónica y la vía alternativa (no canónica), con distintos mecanismos de activación. Los mecanismos oncogénicos en las que participa el factor NF-κB incluyen inflamación crónica, proliferación, apoptosis, angiogénesis, acción sobre células madre del cáncer, metástasis, regulación metabólica y otros mecanismos asociados. En conclusión, existen aún muchas incógnitas sobre los mecanismos y funciones de NF-κB en el contexto celular; el bloqueo completo del factor NF-κB no parece ser una estrategia factible para el tratamiento del cáncer en el momento actual por la diversidad de acciones fisiológicas importantes que se alteran ante su bloqueo. Futuras investigaciones del factor nuclear NF-κB deberían centrarse en la inhibición de la actividad promotora del cáncer, evitando afectar sus funciones fisiológicas normales.
The nuclear factor kappa B (NF-κB) family of transcription factors, which regulates a large range of genes in various immunological and inflammatory response pathways, is of utmost importance. This family consists of five structurally similar members that can activate target genes by attaching to particular regions of deoxyribonucleic acid (DNA). A class of inhibitory proteins usually keep NF-κB proteins in the cytoplasm; however, different oncogenic signaling pathways can activate them and cause malignant phenotypes in the appropriate cells. The main goal of this review article is to understand the regulatory mechanisms of NF-κB transcription factor, its pathogenesis and its potential cancer therapies. From the year 2000 to December 2022, several databases, including PubMed, Scopus and SciELO, were consulted. Finally, the review was developed by searching bibliographic references looking for the keywords related to NF-κB and cancer. The NF-κB transcription factor plays a key role in numerous cell signaling pathways, is involved in a number of biological functions, and its mutations have been linked to cancer and immunological disorders, among other pathologies. Since NF-κB is expressed in all cell types and tissues, many oncogenic mutations can activate NF-κB in tumor cells, opening up new research possibilities for the treatment of cancer. The canonical pathway and the alternative (non-canonical) pathway are two distinct NF-κB signaling pathways with various activation methods. NF-κB is involved in a variety of oncogenic pathways, including chronic inflammation, proliferation, apoptosis, angiogenesis, effect on cancer stem cells, metastasis, metabolic control and other related mechanisms. In conclusion, there are still many unanswered questions regarding the mechanisms and functions of NF-κB in the cellular context. A complete blockade of NF-κB does not appear to be a feasible strategy for the treatment of cancer at this time due to the variety of significant physiological actions that are altered by its blockade. Future research on NF-κB should focus on preventing cancer promotion while preserving the body's natural physiological processes.
摘要
Objective:To evaluate the effect of lipoxin A4 on lipopolysaccharide (LPS)-induced activation of microglia and role of silent information regulator sirtuin 1 (SIRT1)/NF-κB signaling pathway.Methods:This experiment was performed in two parts.PartⅠ The well-growth BV2 microglia were divided into 4 groups ( n=30 each) using a random number table method: control group (group C), LXA4 group (group LXA4), LPS group (group LPS) and LPS+ LXA4 group (group LLI). PartⅡ The well-growth BV2 microglia were divided into 2 groups ( n=30 each) using a random number table method: LPS+ LXA4 group (group LL2) and LPS+ LXA4+ SIRT1 inhibitor EX527 group (group LLE). Cells in group C were commonly cultured without any treatment. In LXA4 group and LPS group, LXA4 (final concentration 100 nmol/L) and LPS (final concentration 100 ng/ml) were added respectively, and then the cells were incubated for 24 h. In LL1 group and LL2 group, LXA4 (final concentration 100 nmol/L) was added at 1 h before treatment with LPS, and the other treatments were similar to those previously described in LPS group. EX527 (final concentration of 5 μmol/L) was added at 30 min before treatment with LXA4, and the other treatments were similar to those previously described in LL2 group.The expression of inducible nitricoxide synthase (iNOS), CD32, arginine synthase 1 (Arg-1), CD206, interleuckin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α) and IL-10 mRNA was detected by real-time polymerase chain reaction. The concentrations of IL-1β, IL-6, TNF-α and IL-10 in the supernatant were measured using enzyme-linked immunosorbent assay. The content of ROS was detected by DCFH-DA. The activity of SOD was measured by WST-8 assay. The expression of NADPH oxidase 2 (NOX2), superoxide dismutase 1 (SOD1), heme oxygenase-1 (HO-1), SIRT1 and acetylated NF-κB p65 was detected by Western blot. Results:Compared with group C, the expression of iNOS, CD32, IL-1β, IL-6 and TNF-α mRNA was significantly up-regulated, the concentrations of IL-1β, IL-6 and TNF-α in the supernatant were increased ( P<0.05), no significant change was found in the expression of Arg-1, CD206 and IL-10 mRNA and IL-10 concentrations in the supernatant, the expression of NOX2 and HO-1 was up-regulated, SOD1 expression was down-regulated, the activity of SOD was decreased, the content of ROS was increased, the expression of SIRT1 was down-regulated, and the expression of acetylated NF-κB p65 was up-regulated in group LPS ( P<0.05). Compared with group LPS, the expression of iNOS, CD32, IL-1β, IL-6 and TNF-α mRNA was significantly down-regulated, the expression of Arg-1, CD206 and IL-10 mRNA was up-regulated, concentrations of IL-1β, IL-6 and TNF-α in the supernatant were decreased, the concentration of IL-10 was increased, the expression of NOX2 was down-regulated, the expression of HO-1 and SOD1 was up-regulated, the activity of SOD was increased, the content of ROS was decreased, the expression of SIRT1 was up-regulated, and the expression of acetylated NF-κB p65 was down-regulated in group LL1 ( P<0.05). Compared with group LL2, the concentrations of IL-1β, IL-6 and TNF-α in the supernatant were significantly increased, the activity of SOD was decreased, the content of ROS was increased ( P<0.05), and no significant change was found in the IL-10 concentration in the supernatant in group LLE ( P>0.05). Conclusions:LXA 4 can inhibit LPS-induced polarization of microglia to M1 phenotype, and the mechanism may be related to enhancement of SIRT1 activity and inhibition of NF-κB transcriptional activity.
摘要
Objective:To evaluate the effect of verbascoside on cold ischemia-reperfusion injury following heterotopic heart transplantation in mice and the relationship with nuclear factor kappa B (NF-κB)/nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3 (NLRP3) signaling pathway.Methods:This experiment was performed in two parts. Part Ⅰ animal experiment Eighteen SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 24-28 g, were divided into 3 groups ( n=6 each) by the random number table method: control group (C group), cold ischemia-reperfusion (I/R) group (I/R group) and cold I/R + verbascoside group (I/R+ VB group). Cold I/R injury model of mouse heart transplantation was prepared by neck heterotopic heart transplantation using the modified non-suture cuff technique. The donor heart in group C was immediately transplanted to the recipient after removal, while the donor heart in group I/R was stored in the 4 ℃ University of Wisconsin solution for 8 h before transplantation to the recipient, and verbascoside 20 mg/kg was intraperitoneally injected at 3 days before surgery in donor and recipient mice in I/R+ VB group. At the end of reperfusion, the myocardial tissue of the transplanted heart was obtained after assessing the beating score for determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity by enzyme-linked immunosorbent assay. Part of the donor myocardium was taken for examination of the pathological results. The expression of NF-κB, p-NF-κB, NOD-like receptor protein 3 (NLRP3), ACS and caspase-1 was detected by Western blot. The expression of IL-1β, TNF-α and IL-6 mRNA was detected by real-time polymerase chain reaction. Part Ⅱ cell experiment Rat cardiomyocyte H9c2 cold hypoxia-reoxygenation model was developed, and the cells were divided into 3 groups ( n=24 each) by the random number table method: control group (C group), cold hypoxia-reoxygenation group (H/R group), and cold hypoxia-reoxygenation+ verbascoside group (H/R+ VB group). The cells were exposed to hypoxia for 18 h at 10 ℃ followed by restoration of reoxygenation for 24 h at 37 ℃ to develop the cold hypoxia-reoxygenation model. The cell viability and LDH activity were determined. The expression of NF-κB and NLRP3 was detected by Western blot, and the expression of phosphorylated NF-κB (p-NF-κB) was detected by immunofluorescence staining. Results:Part Ⅰanimal experiment Compared with C group, the MDA content was significantly increased, the beating score of grafts and SOD activity were decreased, and the expression of p-NF-κB, NLRP3, ACS and caspase-1 and IL-1β, TNF-α and IL-6 mRNA was up-regulated ( P<0.05), and myocardial histopathological injury was aggravated in I/R group. Compared with I/R group, the content of MDA was significantly decreased, the beating score of grafts and SOD activity were increased, the expression of p-NF-κB, NLRP3, ACS and caspase-1 and IL-1β, TNF-α and IL-6 mRNA was down-regulated ( P<0.05), and the myocardial histopathological injury was alleviated in I/R+ VB group. Part Ⅱ cell experiment Compared with C group, the cell viability was significantly decreased, the activity of LDH was increased, and the expression of p-NF-κB, NLRP3, ACS, caspase-1 and p-NF-κB was up-regulated in H/R group ( P<0.05). Compared with H/R group, the cell viability was significantly increased, the activity of LDH was decreased, and the expression of p-NF-κB, NLRP3, ACS, caspase-1 and p-NF-κB was down-regulated in H/R+ VB group ( P<0.05). Conclusions:Verbascoside can alleviate cold I/R injury following heterotopic heart transplantation in mice, and the mechanism may be related to inhibition of activation of NF-κB/NLRP3 signaling pathway.
摘要
Objective:To evaluate the relationship between the mechanism underlying electroacupuncture-induced reduction of postoperative cognitive dysfunction and microglial nuclear factor kappa B (NF-κB)/NOD-like receptor protein 3 (NLRP3) signaling pathway in aged mice.Methods:Sixty SPF healthy male C57BL/6 mice, aged 18 months, weighing 27-30 g, were allocated into 4 groups ( n=15 each) using a random number table method: control group (C group), operation group (O group), operation plus sham electroacupuncture group (O+ SE group) and operation plus electroacupuncture group (O+ Egroup). In O+ E group, the mice received electrical stimulation at the Baihui and Shenting acupoints for a duration of 30 min. The electroacupuncture entailed the utilization of a continuous wave with a frequency of 10 Hz, an intensity of 1 mA, and a daily application for 5 consecutive days. The mice in Y group underwent the same procedure as O+ E group, except that no electrical stimulation was administered. Laparotomy was performed on the mice following a 3% isoflurane anesthesia in O, O+ SE, and O+ E groups. The open field test was conducted in the morning of 6th day after surgery to assess the spontaneous activity and anxiety-like behavior. The novel object recognition test was subsequently performed in the afternoon of 6th day after surgery. The fear conditioning test was performed on 7th day postoperatively. The mice were sacrificed under deep anesthesia at the end of behavioral testing, and the hippocampal tissue was extracted for determination of the expression of phosphorylated NF-κB p65 (p-NF-κB p65), NLRP3, postsynaptic density protein-95 (PSD-95), and synaptic vesicle protein (SYN) (by Western blot) and the co-staining area of NLRP3 and ionized calcium-binding adapter molecule 1 (Iba1, a specific marker for microglia) (by immunofluorescence staining) and for measurement of the dendritic length and dendritic spine density of neurons in the hippocampal CA1 region (using Golgi staining). Results:No statistically significant differences were observed in terms of movement speed, distance traveled, and time of center stay during the open field test among the four groups ( P>0.05). Compared to C group, the percentage of novel object exploration and discrimination index were significantly decreased, the freezing time was prolonged, the expression of p-NF-κB p65 and NLRP3 in the hippocampal tissue was up-regulated, the co-staining area of NLRP3 and Iba1 was increased, the expression of PSD-95 and SYN was down-regulated, and the dendritic length and dendritic spine density of hippocampal neurons were decreased in O, O+ SE and O+ E groups ( P<0.05). Compared to O group, the percentage of novel object exploration and discrimination index were significantly increased, the freezing time was shortened, the expression of p-NF-κB p65 and NLRP3 in the hippocampal tissue was down-regulated, the co-staining area of NLRP3 and Iba1 was decreased, the expression of PSD-95 and SYN was up-regulated, and the dendritic length and dendritic spine density of hippocampal neurons were increased in O+ E group ( P<0.05), and no significant change was found in each parameter in O+ SE group ( P>0.05). Compared to O+ SE group, the percentage of novel object exploration and discrimination index were significantly increased, the freezing time was shortened, the expression of p-NF-κB p65 and NLRP3 in the hippocampal tissue was down-regulated, the co-staining area of NLRP3 and Iba1 was decreased, the expression of PSD-95 and SYN was up-regulated, and the dendritic length and dendritic spine density of hippocampal neurons were increased in O+ E group ( P<0.05). Conclusions:The mechanism by which electroacupuncture attenuates postoperative cognitive dysfunction is associated with inhibition of activation of NF-κB/NLRP3 signaling pathway in microglial cells and improvement in synaptic plasticity in aged mice.
摘要
Metformin, the first-line treatment of type 2 diabetes, has been also increasingly used in the treatment of some skin diseases, such as psoriasis. This review summarizes therapeutic mechanisms of and clinical research progress in metformin in the treatment of psoriasis, aiming to provide ideas and methods for the treatment of psoriasis, especially psoriasis complicated by metabolic diseases.
摘要
Objective To investigate the effect and mechanism of human umbilical cord mesenchymal stem cell exosomes(Exo)on pulmonary inflammation in chronic obstructive pulmonary disease(COPD)rats.Methods A total of 18 male SD rats were randomly divided into the control group,the COPD group and the COPD + Exo group with 6 rats in each group.COPD rat model was estalished by giving cigarette smoke and lipopolysaccharide(LPS)inhalation in the COPD group and the COPD + Exo group.Rats in the COPD + Exo group were injected with 100 μL of human umbilical mesenchymal stem cell exosome diluent(containing 2×107 exosomes)via tail vein.The control group and the COPD group were given equal amounts of phosphate buffered solution.One week after intervention,rats were killed and the peripheral blood,alveolar lavage fluid(BALF)and lung tissue samples were collected.The histopathological changes in lung tissue of each group were observed using Hematoxylin and Eosin(HE)staining.The number of inflammatory cells in peripheral blood was counted by fully automated hematology analyzer.The serum levels of IL-1β,IL-6 and TNF-α were detected by enzyme-linked immunosorbent assay(ELISA).The expression levels of IL-1β,IL-6 and TNF-α mRNA in lung tissue were detected by qRT-PCR.Western blot assay was used to detect the protein expression levels of TLR4,p-NF-κB p65 and IκBα in lung tissue.Results In the control group,the structure of pulmonary alveolus was intact and regular,while in the COPD group,the structure of alveolus was disordered,some alveolus expanded irregularly,fused into bullae and even ruptured.The alveolar septa were widened and filled with a large number of inflammatory cells.Compared with the COPD group,the COPD+ Exo group had more intact alveolar structure and less infiltration of inflammatory cells in the alveolar septum.The inflammatory cells in peripheral blood decreased.The expression levels of IL-1β,IL-6 and TNF-α in serum and lung tissue were decreased(P<0.05).The expression of TLR4 and IκBα protein in lung tissue were increased,while the expression of p-NF-κb p65 protein decreased(P<0.05).Conclusion Human umbilical cord mesenchymal stem cell exosomes can alleviate pulmonary inflammation in COPD rats by modulating TLR4/NF-κB signaling pathway,thereby playing a lung-protective effect.
摘要
Objective:To evaluate the role of Sirtuin 1/nuclear factor κB (SIRT1/NF-κB) signaling pathway in mild hypothermia-induced promotion of microglial polarization during oxygen-glucose deprivation and restoration (OGD/R).Methods:The well-grown BV2 microglia were divided into 4 groups ( n=36 each) using the random number table method: control group (group C), OGD/R group (group O), mild hypothermia group (group M), and mild hypothermia+ SIRT1 specific inhibitor EX527 group (group ME). Cells in group C were commonly cultured without any treatment. Cells in group O were subjected to 3 h of OGD followed by 21 h of restoration of O 2-glucose supply at 37 ℃. Cells in group M were subjected to 3 h of OGD followed by 21 h of restoration of O 2-glucose supply at 33 ℃. Cells in group ME were co-cultured with inhibitor EX527 (final concentration 5 nmol/L) for 12 h in the medium before OGD/R, and the other procedures were conducted as previously described in group M. The cell survival rate was detected by CCK-8 assay. The levels of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and interleukin-10 (IL-10) in supernatant were detected by enzyme-linked immunosorbent assay. The expression of CD206, CD32, inducible nitric oxide synthase (iNOS) and arginine synthase 1 (Arg-1) mRNA was detected by quantitative real-time polymerase chain reaction. The expression of CD206 and CD32 was detected by immunofluorescent staining. The expression of iNOS, Arg-1, SIRT1, NF-κB p65 (p65) and acetylated NF-κB (Ac-p65) was detected by Western blot. Results:Compared with group C, the cell survival rate was significantly decreased, the concentrations of TNF-α, IL-6 and IL-10 in the supernatant were increased, the expression of CD206, Arg-1, CD32 and iNOS was up-regulated, the expression of SIRT1 was down-regulated, and the Ac-p65/p65 ratio was increased in group O ( P<0.05). Compared with group O, the cell survival rate was significantly increased, the concentrations of TNF-α and IL-6 in the supernatant were decreased, the concentration of IL-10 was increased, the expression of CD206, Arg-1 and SIRT1 was up-regulated, the expression of CD32 and iNOS was down-regulated, and the Ac-p65/p65 ratio was decreased in group M ( P<0.05). Compared with group M, the cell survival rate was significantly decreased, the concentrations of TNF-α and IL-6 in the supernatant were increased, the concentration of IL-10 was decreased, the expression of CD206, Arg-1 and SIRT1 was down-regulated, the expression of CD32 and iNOS was up-regulated, and the Ac-p65/p65 ratio was increased in group ME ( P<0.05). Conclusions:SIRT1/NF-κB signaling pathway is involved in mild hypothermia-induced promotion of microglial polarization during OGD/R.
摘要
Objective:To evaluate the role of Toll-like receptor 4 (TLR4)/nuclear transcription factor κB (NF-κB) signaling pathway in long-term cognitive impairment induced by multiple exposures to sevoflurane anesthesia in neonatal rats.Methods:Seventy-five SPF healthy newborn Sprague-Dawley rats of either sex, aged 6 days, weighing 12-20 g, were divided into 3 groups ( n=25 each) using a random number table method: control group (group C), multiple exposures to sevoflurane anesthesia group (group S) and TLR4 inhibitor plus multiple exposures to sevoflurane anesthesia group (group I+ S). The rats in group S and group I inhaled 3% sevoflurane for 2 h at 6, 7 and 8 days after birth. TLR4 inhibitor TAK-242 10 mg/kg was intraperitoneally injected before each exposure to sevoflurane in group I, and the equal volume of normal saline was given instead in the other two groups. The spontaneous activity was evaluated by open field test on day 29 after birth, and the cognitive function was assessed by Morris water maze test on days 30-34 after birth. After the behavioral test, the blood samples from the abdominal aorta were collected, and then the rats were sacrificed under deep anesthesia to isolate the hippocampal tissues for measurement of the levels of S100β and neuron-specific enolase (NSE) in serum and hippocampal interleukin-1β (IL-1β), IL-6 and tumor necrosis factor α (TNF-α) (by enzyme-linked immunosorbent assay), expression of TLR4, NF-κB p65 and phosphorylated NF-κB p65 (p-NF-κB p65) (by Western blot) and for microscopic examination of the pathological changes of hippocampal CA1 region after HE staining. Results:Compared with group C, the escape latency was significantly prolonged, the number of crossing the original platform was reduced, the TLR4 expression was up-regulated, the ratio of p-NF-κB p65/NF-κB p65 was increased, the levels of serum S100β protein and NSE and hippocampal IL-1β, IL-6 and TNF-α were increased ( P<0.05), and the pathological changes in the hippocampal CA1 region were aggravated in group S. Compared with group S, the escape latency was significantly shortened, the number of crossing the original platform was increased, TLR4 expression was down-regulated, the ratio of p-NF-κB p65/NF-κB p65 was decreased, the levels of S100β and NSE in serum and hippocampal IL-1β, IL-6 and TNF-α were decreased ( P<0.05), and the pathological changes in hippocampal CA1 area were significantly attenuated in group P. Conclusions:The mechanism by which multiple exposures to sevoflurane anesthesia induces long-term cognitive impairment is related to activation of TLR4/NF-κB signaling pathway and increase in hippocampal inflammatory responses in neonatal rats.
摘要
Objective:To evaluate the effect of exosomes derived from bone mesenchymal stem cells (BMSCs-EXO) on the postoperative cognitive function and silent infomation regulator 1 (SIRT1)/ nuclear factor kappa B (NF-κB) signaling pathway in aged mice.Methods:BMSCs-EXO were isolated by differential centrifugation method and then identified. Twenty healthy male C57BL/6 aged mice, aged 18 months, weighing 35-40 g, were divided into 4 groups ( n=5 each) using a random number table method: sham operation group (Sham group), operation group (O group), BMSCs-EXO group and EX527 (SIRT1 inhibitor)group. The abdomen regions were shaved for sterilization without exploratory laparotomy in Sham group. Exploratory laparotomy was performed in O group. BMSCs-EXO 50 μg was injected through the tail vein at 1 h before surgery in BMSCs-EXO group. EX527 5 mg/kg was intraperitoneally injected daily at 1-3 days before surgery, and BMSCs-EXO 50 μg was injected through the tail vein at 1 h before surgery in EX527 group. Morris water maze test was used to evaluate the learning and memory ability for 5 consecutive days staring from the 1st day after surgery. Mice were sacrificed at 1 h after the end of Morris water maze test on day 5 after surgery, and the hippocampal tissues were collected for observation of the pathological changes of hippocampal CA1 region and for determination of the expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-1β mRNA (quantitative real-time polymerase chain reaction) and SIRT1 and NF-κB p65 (by Western blot). Results:Compared with Sham group, the escape latency was significantly prolonged, the times of original platform crossing were decreased, the swimming time spent in the original platform quadrant was shortened, the expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-1β mRNA was up-regulated, the SIRT1 expression was down-regulated, the NF-κB p65 expression was up-regulated ( P<0.05), and the pathological changes of hippocampal tissues in CA1 region were found in O group. Compared with O group, the escape latency was significantly shortened, the times of original platform crossing were increased, the swimming time spent in the original platform quadrant was prolonged, the expression of TNF-α, IL-6 and IL-1β mRNA was down-regulated, the expression of SIRT1 was up-regulated, the expression of NF-κB p65 was down-regulated ( P<0.05), and the pathological changes of hippocampal tissues in CA1 region were significantly attenuated in BMSCs-EXO group ( P<0.05). Compared with BMSCs-EXO group, the escape latency was significantly prolonged, the times of original platform crossing were decreased, the swimming time spent in the original platform quadrant was shortened, the expression of TNF-α, IL-6 and IL-1β mRNA was up-regulated, the SIRT1 expression was down-regulated, the NF-κB p65 expression was up-regulated ( P<0.05), and the pathological changes of hippocampal tissues in CA1 region were accentuated in EX527 group. Conclusions:BMSCs-EXO can improve the postoperative cognitive function in aged mice, and the mechanism may be associated with the activation of SIRT1/NF-κB signaling pathway.
摘要
Objective:To evaluate the role of cold-inducible RNA-binding protein (CIRP) in acute renal injury in a mouse model of myocardial ischemia-reperfusion (I/R) and the relationship with nuclear factor kappa B (NF-κB) signaling pathway.Methods:Twenty-four SPF-grade healthy male C57BL/6 mice, aged 6-8 weeks, with body mass index of 24-28 g, were divided into 3 groups ( n=8 each) using a random number table method: sham operation group (Sham group), myocardial I/R group (I/R group) and myocardial I/R + CIRP-derived peptide C23 group (I/R+ C23 group). The model of myocardial I/R was developed by ligation of the left anterior descending coronary artery for 30 min followed by 120-min reperfusion in anesthetized animals. CIRP-derived peptide C23 8 mg/kg was intraperitoneally injected before myocardial ischemia and reperfusion in I/R+ C23 group, while Sham group was only threaded without ligation. Blood samples were collected from the right internal carotid artery at 120 min of reperfusion for determination of the serum creatine kinase isoenzymes (CK-MB), lactic dehydrogenase (LDH), creatinine (Cr) and blood urea nitrogen (BUN) concentrations. Renal tissues were obtained for examination of the pathological changes, and the tubular injury score was assessed. The expression of NF-κB, phosphorylated NF-κB (p-NF-κB), Nod-like receptor protein 3 (NLRP3), interleukin-1beta (IL-1β) and IL-18 in renal tissues was detected by Western blot. The expression of Toll-like receptor 4 (TLR4), NLRP3, IL-1β, TNF-α and IL-6 mRNA was determined by real-time polymerase chain reaction. Results:Compared with Sham group, the levels of serum CK-MB, LDH, Cr and BUN and renal tubule injury score were significantly increased, the expression of p-NF-κB, NLRP3, IL-1β and IL-18 was up-regulated, the expression of TLR4, NLRP3, IL-1β, TNF-α and IL-6 mRNA was up-regulated ( P<0.05), and the pathological injury to renal tissues was aggravated in I/R group. Compared with I/R group, the serum CK-MB, LDH, Cr, BUN and renal tubular injury score were significantly decreased, and the expression of p-NF-κB, NLRP3, IL-1β and and IL-18 was down-regulated, the expression of TLR4, NLRP3, IL-1β, TNF-α and IL-6 mRNA was down-regulated ( P<0.05), and the pathological injury to renal tissues was alleviated in I/R+ C23 group. Conclusions:CIRP is involved in the process of acute renal injury in a mouse model of myocardial I/R, which is associated with activation of NF-κB signaling pathway and promotion of inflammatory responses.
摘要
Liver fibrosis is a compensatory response in the process of tissue repair after chronic liver injury, and it is also a necessary pathological process in the progression of a variety of chronic liver diseases. In the pathological state, the imbalance between hepatic oxidative system and antioxidant system can lead to the excessive production or insufficient clearance of reactive oxygen species (ROS)/reactive nitrogen species (RNS), which may induce the injury of hepatocytes, expand inflammatory response, and promote the development and progression of liver fibrosis. As a master regulator of oxidative stress and inflammatory response, NF-κB plays a key role in the process of liver fibrosis. Therefore, the cascade interaction between ROS/RNS and the NF-κB signaling pathway plays a guiding role in further clarifying the pathogenesis of liver fibrosis and exploring effective prevention and treatment strategies. This article reviews and discusses the interaction between ROS/RNS and the NF-κB signaling pathway and its important role in the progression of liver fibrosis, so as to provide strategies and references for targeted therapy for liver fibrosis.
摘要
Objective:To observe the effects of Traditional Chinese Medicine (TCM)ultrasound drug permeation electrotherapy device on the inflammatory response of rats with cerebral ischemia, and to provide an experimental basis for the clinical application of TCM ultrasound drug permeation electrotherapy device in the treatment of cerebral ischemia.Methods:A total of 72 SD rats were randomly divided into sham-operation group (12 rats) and modeling group (60 rats). The middle cerebral artery occlusion (MCAO) model was prepared by thread embolism in the model group. The rats were divided into model group, Chinese medicine tablet group, blank tablet + TCM ultrasound drug permeation electrotherapy group (hereinafter referred to as "blank tablet + electrotherapy group"), Chinese medicine tablet + TCM ultrasound drug permeation electrotherapy group (hereinafter referred to as "Chinese medicine tablet + electrotherapy group") and butylphthalide group according to the random number table method, with 12 rats in each group. The corresponding treatment was given continuously for 7 days. The neurological function was scored using Longa method evaluation criteria; TTC staining was used to observe the infarct volume and calculate the percentage of infarct volume; HE staining was used to observe the cell morphology of cortical area in each group of rats; ELISA was used to detect the serum TNF-α and IL-1β levels in each group of rats; TLR4, MyD88 and NF-κBp65 protein expressions in hippocampal tissue of each group of rats on the infarct side were detected by Western blot method.Results:Compared with the model group, the neurological function scores of rats in the blank tablet + electrotherapy group, the herbal tablet + electrotherapy group, and the butylphthalein group significantly decreased ( P<0.05), the percentage of cerebral infarct volume significantly decreased ( P<0.05), the contents of serum TNF-α and IL-1β significantly decreased ( P<0.05), and the expressions of TLR4 (0.42±0.07, 0.31±0.07, 0.19±0.04 vs. 0.68±0.14), MyD88 (0.39±0.12, 0.30±0.07, 0.23±0.11 vs. 0.67±0.10), NF-κBp65 (0.32±0.03, 0.27±0.02, 0.17±0.03 vs. 0.57±0.12) protein in hippocampal tissue significantly decreased ( P<0.05). Conclusion:The TCM ultrasound drug permeation electrotherapy device can inhibit TLR4, MyD88, NF-κBp65 protein expressions and reduce the release of serum inflammatory factors TNF-α and IL-1β, thus exerting cerebral ischemic protective effects.
摘要
Objective:To study the protective effect of Ophiopogonin D on lipopolysaccharide (LPS)-induced mouse macrophage RAW264.7 and its related mechanism.Methods:Mouse macrophage RAW264.7 cells were cultured and divided into normal control group, model group and Ophiopogonin D pretreatment group according to random number table method. The activity of Ophiopogonin D on RAW264.7 cells was detected by CCK-8 method; the protein levels of TNF-α, IL-1β, IL-6, reactive oxygen species (ROS), MDA and SOD were detected by ELISA; the protein expression of NF-κB, TLR4, NF-E2-related factor2 (Nrf2) and heme oxygenase-1 (HO-1) were detected by Western Blot.Results:Compared with model group, the levels of TNF-α, IL-1β, IL-6, ROS and MDA in cell supernatant of Ophiopogonin D group were decreased ( P<0.05), and the level of SOD was increased ( P<0.05). The protein expressions of NF-κB (0.76±0.10 vs. 2.26±0.17) and TLR4 (0.98±0.09 vs. 1.74±0.19) significantly decreased ( P<0.05). The protein expressions of Nrf2 (0.85±0.03 vs. 0.54±0.03) and HO-1 (0.97±0.11 vs. 0.37±0.04) significantly increased ( P<0.05). Conclusion:Ophiopogonin D may reduce inflammatory response by reducing TLR4/NF-κB pathway, and activate Nrf2/HO-1 pathway to reduce oxidative damage and play a protective role.