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1.
Rev. invest. clín ; 74(3): 147-155, May.-Jun. 2022. tab
Article in English | LILACS-Express | LILACS | ID: biblio-1409573

ABSTRACT

ABSTRACT Background: The inflammatory response in gout disease is induced by the activation of NLR family pyrin domain-containing 3 (NLPR3) signaling pathway mediated by IL-1β release. Objective: The objective of the study was to determine the association between single nucleotide polymorphisms (SNPs) within NLRP3 inflammasome genes and gout susceptibility. Methods: Mexican patients with gout from the National Rehabilitation Institute and General Hospital of Mexico were enrolled. A healthy control group was also included. We analyzed the frequency and allelic distribution of eight SNPs from seven different genes within the NLRP3 inflammasome signaling pathway: TLR4 rs2149356, CD14 rs2569190, NLRP3 rs3806268, NLRP3 rs10754558, CARD8 rs2043211, IL-1β rs1143623, P2RX7 rs3751142, and PPARGC1B rs45520937 SNPs. Results: We found that the SNP rs45520937 of PPARGC1B was associated with the risk of developing gout when it was analyzed using the dominant model (Odds ratio [OR] = 2.30; 95% confidence interval [CI]: 1.09-4.86; p = 0.030), and it is proposed that the adaptor molecule CD14 rs2569190 polymorphism could be associated with a lower risk of gout under an additive model (OR= 0.41;95% CI: 0.16-1.05; p = 0.064). No significant associations were identified for the remaining SNPs. Conclusion: Our findings suggest that the PPARGC1B rs45520937 SNP is associated with gout susceptibility.

2.
Int. j. morphol ; 40(1)feb. 2022.
Article in English | LILACS-Express | LILACS | ID: biblio-1385582

ABSTRACT

SUMMARY: Skeletal muscle injury is an acute inflammatory condition caused by an inflammatory response. To reduce inflammatory cell infiltration and relieve skeletal muscle injury, efficient treatment is urgently needed. Nitric oxide is a free radical molecule reported to have anti-inflammatory effects. In this study, we showed that NO could inhibit the inflammatory response of C2C12 cells in vitro and protect rat skeletal muscle injury from notexin in vivo. NO synthase inhibitor (L-NG-Nitroarginine Methyl Este?L-NAME) and NO donor (sodium nitroprusside dehydrate ?SNP) were used to explore the vital role of lipopolysaccharides (LPSs) in LPS-stimulated C2C12 myoblasts.The expression of IL-18 and IL-1b was upregulated by L-NAME and downregulated by SNP, as indicated by the ELISA results. NO can reduce ASC, Caspase-1, and NLRP3 mRNA and protein levels. Furthermore, NO was detected in the rat model. The results of immunohistochemical staining showed that the production of DMD decreased. We conducted qRT-PCR and western blotting to detect the expression of Jo-1, Mi-2, TLR2, and TLR4 on day 6 post injury following treatment with L-NAME and SNP. The expression of Jo-1, Mi-2, TLR2, and TLR4 was upregulated by L-NAME and significantly reversed by SNP. NO can alleviate C2C12 cell inflammatory responses and protect rat skeletal muscle injury from notexin.


RESUMEN: La lesión del músculo esquelético es una afección inflamatoria aguda causada por una respuesta inflamatoria. Para reducir la infiltración de células inflamatorias y aliviar la lesión del músculo esquelético es necesario un tratamiento eficaz. El óxido nítrico es una molécula de radicales libres que tiene efectos antiinflamatorios. En este estudio, demostramos que el ON podría inhibir la respuesta inflamatoria de las células C2C12 in vitro y proteger la lesión del músculo esquelético de rata de la notexina in vivo. El inhibidor de ON sintasa (L-NG-nitroarginina metil este, L-NAME) y el donante de ON (nitroprusiato de sodio deshidratado, SNP) se utilizaron para explorar el papel vital de los lipopolisacáridos (LPS) en los mioblastos C2C12 estimulados por LPS. La expresión de IL- 18 e IL-1b fue regulada positivamente por L-NAME y regulada negativamente por SNP, como indican los resultados de ELISA. El ON puede reducir los niveles de proteína y ARNm de ASC, Caspasa-1 y NLRP3. Además, se detectó ON en el modelo de rata. Los resultados de la tinción inmunohistoquímica mostraron que disminuyó la producción de DMD. Realizamos qRT-PCR y transferencia Western para detectar la expresión de Jo-1, Mi-2, TLR2 y TLR4 el día 6 después de la lesión después del tratamiento con L-NAME y SNP. La expresión de Jo-1, Mi-2, TLR2 y TLR4 fue regulada positivamente por L- NAME y significativamente revertida por SNP. El ON puede aliviar las respuestas inflamatorias de las células C2C12 en ratas, y proteger la lesión del músculo esquelético de la notexina.

3.
Article in Chinese | WPRIM | ID: wpr-935785

ABSTRACT

Objective: To explore the mechanism of reactive oxygen species/thioredoxin-interacting protein/nucleotide-binding oligomerization domain-like receptor 3 (ROS/TXNIP/NLRP3) pathway in the skin injury of trichloroethylene (TCE) sensitized mice. Methods: In August 2020, 40 female BALB/c mice were randomly divided into control group (n=5) , solvent control group (n=5) , TCE treatment group (n=15) and TCE+(2-(2, 2, 6, 6-Tetrameyhylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl) triphenylphosphonium chloride (Mito TEMPO) treatment group (n=15) . The TCE sensitization model was established. Mice in the TCE treatment group and TCE+Mito TEMPO treatment group were divided into the sensitized positive group and the sensitized negative group according to the skin erythema and edema reactions on the back of the mice 24 h after the last stimulation. The mice were sacrificed 72 h after the last stimulation, the back skin of the mice was taken, and the skin lesions were observed. Immunohistochemistry (IHC) was used to detect the expression level of NLRP3, and the Western Blot was performed to detect the expression levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) , cysteinyl aspartate specific proteinase 1 (Caspase 1) , Interleukin-1β (IL-1β) and TXNIP proteins in the skin of the mice, the reactive oxygen species (ROS) kit was used to detect the level of intracellular ROS in the back skin tissue. Results: The sensitization rates of TCE treatment group and TCE+Mito TEMPO treatment group were 40.0% (6/15) and 33.3% (5/15) , respectively, and there was no significant difference between the two groups (P>0.05) . The back skin of the mice in the TCE sensitized positive group was thickened and infiltrated by a large number of inflammatory cells. The number of mitochondria in the epidermis cells was significantly reduced, the mitochondrial crest disappeared and vacuolar degeneration occurred. TCE+Mito TEMPO sensitized positive group had less damage, more mitochondria and relatively normal cell structure. Compared with the solvent control group and corresponding sensitized negative groups, the expression levels of NLRP3, ASC, Caspase 1, IL-1β, TXNIP proteins and the content of ROS in the TCE sensitized positive group and TCE+Mito TEMPO sensitized positive group were significantly increased (P<0.05) . Compared with TCE sensitized positive group, the expression levels of NLRP3, ASC, Caspase 1, IL-1β, TXNIP proteins and the content of ROS in the TCE+Mito TEMPO sensitized positive group were significantly decreased (P<0.05) . Conclusion: ROS/TXNIP/NLRP3 pathway was activated and then encouraged the release of IL-1β, finally aggravated the TCE-induced skin injury.


Subject(s)
Animals , Carrier Proteins , Caspase 1/metabolism , Female , Inflammasomes/metabolism , Mice , Mice, Inbred BALB C , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Reactive Oxygen Species/metabolism , Solvents , Thioredoxins/metabolism , Trichloroethylene/toxicity
4.
Article in Chinese | WPRIM | ID: wpr-934029

ABSTRACT

SARS-CoV-2, the pathogen of the COVID-19 pandemic, causes serious damage to human health and social stability. In severe COVID-19 cases, the infection triggers cytokine storm, resulting in multi-organ excessive inflammatory responses and even failure, which eventually leads to death. Recent studies have shown the activation of nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3 (NLRP3) inflammasome plays an essential role in the pathogenesis of COVID-19. SARS-CoV-2 can activate NLRP3 inflammasome through several pathways, thereby inducing the release of a large number of pro-inflammatory cytokines. This article reviews the activation of NLRP3 inflammasome caused by SARS-CoV-2 infection and the possible molecular mechanisms, and summarizes the progress in targeted inhibition of NLRP3 inflammation, aiming to provide a new strategy for the treatment of SARS-CoV-2 infection.

5.
Article in Chinese | WPRIM | ID: wpr-934027

ABSTRACT

Juvenile idiopathic arthritis (JIA) is a heterogeneous group of diseases characterized by chronic inflammatory arthritis of unknown cause, lasting six weeks or longer, and accompanied by organ damages. It is the most common chronic inflammatory rheumatic disease in childhood with unclear aetiology. A20, a protein encoded by the tumor necrosis factor α-induced protein 3 gene (TNFAIP3), regulates cell inflammatory response and apoptosis through suppressing inflammatory NF-κB signaling by acting as an ubiquitin-editing enzyme. NOD-like receptor protein 3 (NLRP3) inflammasome, a multiprotein complex formed by a subgroup of intracellular pattern recognition receptors, mediates the activation of caspase-1 and the secretion of proinflammatory cytokines IL-1β and IL-18 in response to microbial infection and cellular damage. A20 could directly reduce the basal expression of NLRP3 to impair caspase-1 activation and inhibit the assembling of NLRP3 inflammasome by suppressing the activation of NF-κB, playing a crucial anti-inflammatory role in JIA. A20 and NLRP3 inflammasome may be promising prognostic markers and therapeutic targets in JIA. This review summarized the structure and biological function of A20 and NLRP3 inflammasome and analyzed their roles in the genetic susceptibility and pathogenesis of JIA.

6.
Chinese Journal of Nephrology ; (12): 344-351, 2022.
Article in Chinese | WPRIM | ID: wpr-933866

ABSTRACT

Objective:To investigate the level of endogenous hydrogen sulfide (H 2S) in contrast-induced acute kidney injury (CIAKI), as well as the potential role of H 2S against CIAKI by down-regulating NLRP3 inflammasome. Methods:Twenty-four healthy male Sprague-Dawley rats, weighing 180-220 g, were randomly divided into three groups according to the random number table method: control group, CIAKI group (iopromide 2.9 g/kg) and CIAKI+NaHS group (NaHS 4 mg/kg for three days before 2.9 g/kg iopromide injection). Kidneys were collected for whole-genome sequencing and bioinformatic analysis. HE and PAS staining were used for kidney histological examination. TUNEL assays were applied to detect renal tubular epithelial injury. Expressions of NLRP3 inflammasome (NLRP3, ASC and caspase-1) were evaluated by immunofluorescence staining. The role of H 2S in contrast (iopromide 200 mgI/kg)-induced injury on human renal tubular epithelium (HK-2 cells) was investigated, and CCK-8 assay was used to detect cellular viability. Results:Compared with the control group, the expression of endogenous H 2S synthetases-related genes [cystathionine β-synthase ( CBS), cystathionine-γ-lyase ( CSE) and 3-mercaptopyruvate sulfurtransferase ( 3- MST)] was lower in CIAKI group (all P<0.05). The gene expression levels of CBS, CSE and 3- MST were negatively correlated with renal function biomarkers serum creatinine, blood urea nitrogen and cystatin-C (all P<0.05). Compared with the CIAKI group, CIAKI+NaHS group showed alleviated creatinine, blood urea nitrogen and cystatin-C, improved histological changes, reduced apoptosis. Moreover, the expression levels of NLRP3, ASC and caspase-1 in CIAKI+NaHS group were lower than those in CIAKI group (all P<0.05). In HK-2 cells, compared with the contrast group, the cellular viability was higher in the contrast+NaHS group; reducing endogenous H 2S by CBS inhibitor could enhance contrast-induced cell viability ( P<0.05). Conclusions:Injury of endogenous H 2S system is pivotal to CIAKI pathogenesis. Up-regulation of H 2S ameliorates renal injury of CIAKI rats, which may be related to regulation of NLRP3 inflammasome.

7.
Article in Chinese | WPRIM | ID: wpr-933366

ABSTRACT

Objective:To investigate the effect of high glucose on the release of interleukin (IL)-1β and IL-18 in placental trophoblast by activating NLRP3 inflammasome.Methods:Gestational diabetes mellitus(GDM) placentas and control placentas were collected and the expression levels of NLRP3 and Caspase-1 were determined. Human placental trophoblast HTR-8/SVneo were cultured and divided into control group(5.5 mmol/L glucose), high glucose group(25 mmol/L glucose), DMSO+ high glucose group, and Ac-YVAD-cmk(NLRP3 inflammasome inhibitor)+ high glucose group. The expression levels of NLRP3 and Caspase-1 in cells as well as the contents of IL-1β and IL-18 in the medium were determined.Results:The expression levels of NLRP3 and Caspase-1 in GDM placenta were higher than those in control placenta( P<0.05) and positively correlated with homeostasis model assessment of insulin resistant index(HOMA-IR) and fasting insulin. The expression levels of NLRP3 and Caspase-1 in HTR-8/SVneo cells and the secretion levels of IL-1β and IL-18 in high glucose group were higher than those in control group( P<0.05). Ac-YVAD-cmk significantly suppressed high glucose-stimulated IL-1β and IL-18 secretion( P<0.05). Conclusion:High glucose promotes the release of IL-1β and IL-18 from placental trophoblast via activating NLRP3 inflammasome.

8.
Article in Chinese | WPRIM | ID: wpr-931089

ABSTRACT

Objective:To investigate the inhibitory effect of melatonin on pyroptosis of lens epithelium cells (HLECs) induced by hydrogen peroxide (H 2O 2) and its mechanism. Methods:The cultured HLECs were divided into normal control group, model control group, melatonin group, vitamin E group, and vitamin E solvent group.Cells in melatonin group, vitamin E group and vitamin E solvent group were pre-cultured with 1×10 -6 mol/L melatonin, 100 μmol/L vitamin E or equal volume of vitamin E solvent, then cultured with 100 μmol/L H 2O 2, respectively, and the cells in the normal control group and model control group were cultured with normal condition or 100 μmol/L H 2O 2, respectively.The HLECs transfected with nuclear factor erythroid 2-related factor 2 short hairpin RNA (shNrf2) or shNrf2 negtive control lentivirus and following with 100 μmol/L H 2O 2 treatment were served as shNrf2 group and shNrf2 negative control group, respectively; and the transfected cells treated with 1×10 -6 mol/L melatonin and subsequent 100 μmol/L H 2O 2 treatment were served as melatonin+ shNrf2 group and melatonin+ shNrf2 negative control group.The activity of lactic dehydrogenase (LDH) and the expression levels of interleukin (IL)-1β and IL-18 in the culture supernatant were detected by the enzyme linked immunosorbent assay (ELISA) kit.The concentration of reactive oxygen species (ROS) was assessed by flow cytometry.The expression level of Nrf2, NLRP3, apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain (ASC), caspase-1 p20 and GSDMD-N proteins were evaluated by Western blot. Results:Compared with model control group, the activity of LDH and the concentrations of IL-1β and IL-18 were significantly decreased in melatonin group and vitamin E group, showing statistically significant differences (all at P<0.05). The ROS fluorescence intensities were 13 040.67±1 550.66 and 12 593.67±1 677.06 in melatonin group and vitamin E group, respectively, which were significantly lower than 18 310.33±1 248.01 in model control group (both at P<0.05). The relative expression levels of Nrf2 protein were 4.24±0.44 and 3.73±0.38 in melatonin group and vitamin E group, respectively, which were significantly higher than 2.28±0.34 in model control group, and the relative expression levels of NLRP3, ASC, caspase-1 p20 and GSDMD-N in melatonin group and vitamin E group were significantly decreased than model control group (all at P<0.05). The relative expression level of Nrf2 protein in shNrf2 group and melatonin+ shNrf2 group was significantly reduced, and the expression levels of LDH, IL-1β, IL-18, ROS content, NLRP3, ASC, caspase-1 p20 and GSDMD-N were significantly increased in comparison with shNrf2 negative control group and melatonin+ shNrf2 negative control group, respectively (all at P<0.05). Conclusions:Melatonin can inhibit the release of NLRP3 inflammasome by activating Nrf2, and has an inhibitory effect on the pyroptosis of HLECs.

9.
Article in Chinese | WPRIM | ID: wpr-930215

ABSTRACT

Objective:To investigate the effect of curcumin on the nucleotide-binding oligomerization domain-like receptor protein (NLRP3) inflammasome in cardiomyocytes of rats with early sepsis and its mechanism.Methods:Twenty-four male SD rats were randomly divided into three groups: the sham-operated group (sham group, n=8), the sepsis group ( n=8), and the curcumin intervention group (Cur group, n=8). A rat model of sepsis was prepared by cecal ligation and perforation (CLP). After modeling, 100 mg/kg of curcumin was intraperitoneally injected and repeated 24 h later. Rats in the sepsis group were injected with normal saline. The levels of myocardial injury-specific troponin T (cTnT) in rat plasma were detected by ELISA at 6, 24, and 48 h. Hematoxylin-eosin (HE) staining was used to observe the pathological injury of the myocardium in the myocardial tissue of rat at 48 h. The apoptosis of cardiomyocytes was detected by TUNEL assay. The expression levels of Cleaved caspase-1, NLRP3 and IL-1β protein were detected by Western blot, and the ultrastructural changes of the cardiomyocytes were observed under a transmission electron microscope. Results:The levels of cTnT in rat plasma at 6, 24, and 48 h in the Cur group were significantly lower than those in the sepsis group ( P<0.05). HE staining showed infiltration, cell edema and necrosis of myocardial inflammatory cells in the sepsis group, while only partial cell edema and necrosis were observed in the Cur group. TUNEL assay showed that the apoptosis of cardiomyocytes in the Cur group was significantly lower than that in the sepsis group [(28.4±2.3)% vs. (43.6±3.8)%, P<0.05]. The expression levels of Cleaved caspase-1, NLRP3 and IL-1β in the Cur group were lower than those in the sepsis group and higher than those in the sham-operated group ( P<0.05). Under transmission electron microscopy, the nuclei in the sham-operated group had intact membranes and uniform chromatin distribution; in the sepsis group chromatin margination, pyknosis, mitochondrial cristae breakage, cavitation, and partial breakage of sarcomere were observed; while in the Cur group partial chromatin margination, slightly edema and dilation of mitochondria, with basically complete morphology were observed. Conclusions:Curcumin inhibits NLRP3-mediated acute myocardial injury in septic rats, and its mechanism may be related to pyroptosis induced by the down-regulation of the expression of Cleaved caspase-1 and IL-1β protein.

10.
Article in English | WPRIM | ID: wpr-929249

ABSTRACT

Nephrotic syndrome (NS) is a kidney disease characterized by hypertriglyceridemia, massive proteinuria, hypo-albuminemia and peripheral edema. Sinkihwan-gamibang (SKHGMB) was recorded in a traditional Chinese medical book named "Bangyakhappyeon ()" and its three prescriptions Sinkihwan, Geumgwe-sinkihwan, and Jesaeng-sinkihwan belong to Gamibang. This study confirmed the effect of SKHGMB on renal dysfunction in an NS model induced by puromycin aminonucleoside (PAN). The experimental NS model was induced in male Sprague Dawley (SD) rats through injection of PAN (50 mg·kg-1)via the femoral vein. SKHGMB not only reduced the size of the kidneys increased due to PAN-induced NS, but also decreased proteinuria and ascites. In addition, SKHGMB significantly ameliorated creatinine clearance, creatinine, and blood urea nitrogen. SKHGMB relieved glomeruli dilation and tubules fibrosis in the glomeruli of the NS model. SKHGMB inhibited the protein and mRNA levels of the NLRP3 inflammasome including NLRP3, ASC, and pro-caspase-1 in NS rats. SKHGMB reduced the protein and mRNA levels of fibrosis regulators in NS rats. The results indicated that SKHGMB exerts protective effects against renal dysfunction by inhibiting of renal inflammation and fibrosis in NS rats.


Subject(s)
Animals , Kidney , Male , Nephrotic Syndrome/drug therapy , Proteinuria/metabolism , Puromycin Aminonucleoside/toxicity , Rats , Rats, Sprague-Dawley
11.
Acta Pharmaceutica Sinica ; (12): 2612-2621, 2022.
Article in Chinese | WPRIM | ID: wpr-941520

ABSTRACT

More and more studies have shown that NOD-like receptor protein 3 (NLRP3) inflammasome has become the regulatory factor of inflammatory response and protective immunity, and the assembly and activation of NLRP3 inflammasomes are closely related to the anti-tumor immunity effect. Depending on the cell type and stimuli, activation of the NLRP3 inflammasome can induce immune cells to become polarized, hyperactive, or pyroptotic, releasing interleukin (IL)-1β and IL-18, which leads to cascade immune or inflammatory responses, and its role in tumor immunity has received extensive attention. Here, we review the mechanisms of the NLRP3 inflammasome enhancing CD8+ T cells-mediated anti-tumor immunity by inducing the pyroptosis of tumor cell, the pyroptosis or hyperactive state of dendritic cells (DCs), and the pyroptosis or polarization of the macrophages. Different anti-tumor immune roles of NLRP3 inflammasome activation in tumor cells and immune cells provide new directions for future research and may influence the development of next-generation immunotherapy.

12.
Acta Pharmaceutica Sinica ; (12): 2743-2750, 2022.
Article in Chinese | WPRIM | ID: wpr-941500

ABSTRACT

Erhuang quzhi compounds is one of the protecting liver and inhibiting toxin prescriptions series summarized by Jinqi Yuan and other famous doctors of traditional Chinese medicine during the long-term clinical practice. It is very effective for non-alcoholic fatty liver disease (NAFLD), but its mechanism is not clear. This research investigated mechanism of Erhuang quzhi granules (EQG) in the treatment of NAFLD. All the animal welfare and experimental procedures are in accordance with the regulations of the Animal Ethics Committee of the First Affiliated Hospital of Shihezi University. Mouse models of NAFLD were established by feeding with methionine and choline deficient diet (MCDD) for five weeks. While feeding MCDD, the treatment groups were given EQG (16.25 g·kg-1·d-1) and atorvastatin (ATO, 7.20 mg·kg-1·d-1) by gavage. The effects of EQG on serum biochemical indices, liver pathological changes, and inflammatory cytokines in mice of NAFLD were investigated. Quantitative real-time PCR (qPCR), immunocytochemistry (ICH) and Western blot assays were used to detect the levels of mRNA and protein associated with nuclear factor kappa B/Nod-like receptor protein 3 (NF-κB/NLRP3) in liver. The results showed that EQG significantly reduced the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), and improved the level of low-density lipoprotein cholesterol (LDL-C). The result of hematoxylin-eosin (HE) staining showed that EQG reduced lipid deposition in livers of mice. Meanwhile, EQG notably decreased the levels of interleukin (IL)-1β, IL-6, IL-18 and tumor necrosis factor-α (TNF-α), and mRNA levels of NF-κB, NLRP3, IL-1β, TNF-α, down-regulated the expression of F4/80, IκB kinase β (IKKβ), NLRP3 and apoptosis-associated speck-like protein containing a CARD (ASC) and inhibited the activation of NF-κB and cysteinyl aspartate specific proteinase-1 (caspase-1). These findings announced that EQG could improve NAFLD via NF-κB/NLRP3 pathway possibly, which provides a theoretical basis for the further development and utilization of EQG in clinic.

13.
Article in Chinese | WPRIM | ID: wpr-940830

ABSTRACT

ObjectiveTo explore the effects of phillygenin (PHI) on the inflammation in L02 cells induced by lipopolysaccharide (LPS) and adenosine triphosphate (ATP) and the expression of purinergic 2X7 receptor (P2X7R), NOD-like receptor family pyrin domain containing 3 (NLRP3), and nuclear factor kappa B (NF-κB) expression. MethodIn this study, the inflammation model was induced in L02 cells by 100 μg·L-1 LPS treatment for 24 h and 5 mmoL·L-1 ATP treatment for 5 h. The cells in the PHI groups were cultured with PHI (100, 50, 25 mg·L-1) for 6 h in the LPS treatment period, followed by LPS treatment for another 18 h. After ATP treatment for 5 h, the mRNA and protein expression of interleukin-1β (IL-1β), interleukin-18 (IL-18), P2X7R, NLRP3, Caspase-1 precursor (pro-Caspase-1), cleaved Caspase-1, NF-κB, and NF-κB inhibitor protein α (IκBα) in L02 cells was detected by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively. Molecular docking was used to predict whether P2X7R could bind to PHI, and DCFH-DA was employed to detect the accumulation of reactive oxygen species (ROS) in cells. P2X7R was silenced by small interfering ribonucleic acid (siRNA), and then the mRNA expression of IL-1β, IL-18, P2X7R, NLRP3, Caspase-1, NF-κB, and IκBα was detected by Real-time PCR. ResultReal-time PCR and Western blot showed that compared with the normal group, the model group showed increased expression of IL-1β and IL-18 (P<0.05), and compared with the model group, the PHI groups showed down-regulated IL-1β, IL-18 mRNA and protein expression (P<0.05). Molecular docking suggested a good binding effect of PHI to P2X7R. Real-time PCR and Western blot analysis showed that the expression of P2X7R in the model group was significantly up-regulated compared with that in the normal group (P<0.05), and compared with the model group, the PHI groups showed down-regulated mRNA and protein expression of P2X7R (P<0.05). DCFH-DA results showed that compared with the normal group, the model group showed increased content of ROS (P<0.05), and compared with the model group, the PHI groups decreased the accumulation of ROS (P<0.05). As demonstrated by Real-time PCR and Western blot, compared with the normal group, the model group showed increased expression of NLRP3 inflammasome and NF-κB (P<0.05), and compared with the model group, the PHI groups significantly decreased the mRNA and protein expression of NLRP3 and cleaved Caspase-1, and up-regulated the mRNA and protein expression of NF-κB and IκBα (P<0.05). Real-time PCR analysis showed that compared with the results in the model group, after silencing P2X7R by siRNA, the mRNA expression of IL-1β, IL-18, P2X7R, NLRP3, Caspase-1, NF-κB, and IκBα was decreased (P<0.05). PHI exerted the same effect, and the mRNA expression was further reduced after the combination of them. ConclusionPHI is presumed to suppress the expression of the NLRP3/NF-κB signaling pathway by down-regulating upstream P2X7R to alleviate the LPS/ATP-induced inflammation in L02 cells, suggesting that P2X7R may be the target of PHI against inflammation.

14.
Article in Chinese | WPRIM | ID: wpr-940825

ABSTRACT

ObjectiveTo explore the intervention effect and mechanism of Buzhong Yiqiwan (BZYQ) on colitis mice. MethodSixty-four C57BL/6 mice were randomly divided into 2 weeks blank group, 2 weeks model group, 2 weeks high-dose BZYQ (12 g·kg-1) group, 2 weeks low-dose BZYQ (6 g·kg-1) group, 4 weeks blank group, 4 weeks model group, 4 weeks high-dose BZYQ (12 g·kg-1) group, and 4 weeks low-dose BZYQ (6 g·kg-1) group. The colitis model was induced in mice by feeding 3% dextran sodium sulfate (DSS) for 7 days. The mice received BZYQ (12 and 6 g·kg-1) by gavage on the 8th day after modeling, once per day, and sacrificed on the 2nd and 4th weeks, correspondingly. The colon length and weight of mice in each group were measured. Hematoxylin-eosin (HE) staining was used for pathological observation and colonic mucosal inflammation was scored. The mRNA and protein expression of NOD-like receptor thermoprotein domain 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), and cysteinyl aspartate-specific protease-1 (Caspase-1) was detected by real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of inflammatory cytokines, such as interleukin (IL)-1β, IL-18, and IL-33 in colonic tissues. ResultCompared with the 2 weeks blank group, the 2 weeks model group showed shortened colon length, increased colon weight (P<0.05), loss of epithelial cells, destruction of gland structure, infiltration of a large number of inflammatory cells in mucosa and submucosa, local crypt abscess, and increase in mucosal inflammation score (P<0.01) as revealed by light microscopy, elevated levels of IL-1β, IL-18, and IL-33 in colonic tissues (P<0.05), and increased mRNA and protein expression of NLRP3, ASC, and Caspase-1 (P<0.05). The intervention of BZYQ (12 g·kg-1) restored colon length, alleviated mucosal injury (P<0.05), down-regulated the content of IL-18 (P<0.05), reduced the mRNA expression of NLRP3 and ASC as well as the protein expression of ASC and Caspase-1 compared with the conditions in the 2 weeks model group. Compared with the 4 weeks blank group, the 4 weeks model group showed decreased colon length, increased colon weight (P<0.05), decreased glands in the mucosal layer, expansion of glandular cavity, atrophy of crypt, local connective tissue hyperplasia and lymphocyte infiltration, increased inflammation score (P<0.01) as revealed by the light microscopy, increased expression of IL-1β, IL-18, and IL-33 (P<0.05), and elevated mRNA and expression of NLRP3 inflammasome complex (P<0.05). Compared with the conditions in the 4 weeks model group, the intervention of BZYQ (12 and 6 g·kg-1) could improve colon length and weight (P<0.05), and the intervention of BZYQ (12 g·kg-1) could also improve the inflammation score of the colon (P<0.05). Different from the acute stage, the intervention of BZYQ (12 and 6 g·kg-1) increased the content of IL-33 in the intestinal mucosa and up-regulated the mRNA and protein expression of NLRP3 inflammasome complexes ASC and Caspase-1 (P<0.05). ConclusionBZYQ can relieve the injury of colitis induced by DSS in mice. The mechanism is related to the regulation of intestinal immune response mediated by NLRP3 inflammasome, and it has different regulatory effects in acute and chronic inflammation stages.

15.
Article in Chinese | WPRIM | ID: wpr-940789

ABSTRACT

ObjectiveTo investigate the mechanism of Chaihu Qinggantang (CHQGT) in the treatment of granulomatous lobular mastitis (GLM) in the rat model. MethodSixty female rats were divided into a normal group, a model group, a prednisolone group (0.001 8 g·kg-1), and three CHQGT low-dose, medium-dose, and high-dose groups (4.5, 8.9, 17.8 g·kg-1). The tissue homogenates mixed with GLM lesion tissue and Fritner's reagent were used for modeling. After modeling, the treatment groups were given corresponding treatment factors, and the normal group and the model group were given the equal volume of normal saline. The changes in mammary gland of rats were observed after 14 d. Hematoxylin-eosin (HE) staining was used to observe the histopathological changes in breast samples. The mRNA expressions of NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome, Caspase-1, and interleukin-1β (IL-1β) were detected by real-time quantitative fluorescence polymerase chain reaction (Real-time PCR). The protein expressions of NLRP3, Caspase-1, IL-1β, and IL-18 were detected by Western bolt. ResultAs compared with the normal group, the breasts of rats in the model group were obviously swelling, and mammary gland inflammation index was significantly increased (P<0.01). Pathological changes included the formation of granuloma centered on the lobule of mammary gland with a large number of inflammatory cells such as lymphocytes and plasma cells. The mRNA expressions of NLRP3, Caspase-1, and IL-1β, and the protein expressions of NLRP3, Caspase-1, IL-1β, and IL18 in the model group were significantly increased (P<0.01). Compared with the model group, the treatment groups improved breast swelling, and the CHQGT medium and high-dose groups and the prednisolone group reduced inflammation index to some extent after treatment (P<0.05, P<0.01). The inflammation degree of mammary gland was significantly improved, and inflammatory cells such as macrophages, lymphocytes, and plasma cells were reduced to varying degrees in pathological aspects. The mRNA expressions of NLRP3, Caspase-1, and IL-1β, and the protein expressions of NLRP3, Caspase-1, IL-1β, and IL-18 in the CHQGT high-dose group and the prednisolone group were significantly down-regulated (P<0.05, P<0.01). ConclusionCHQGT inhibits inflammation and treats GLM in rats. The mechanism is possibly related to the inhibition of NLRP3/IL-1β signaling pathway, which provides a new target for the prevention and treatment of GLM by Qingxiao method.

16.
Article in Chinese | WPRIM | ID: wpr-940785

ABSTRACT

Diabetes mellitus (DM) is a metabolic disease mainly characterized by chronic hyperglycemia and has multiple etiologies. The complications of DM, such as coronary atherosclerosis, nephropathy, foot disease and cardiac dysfunction, have high morbidity, disability rate and mortality. DM and its complications have a long course of disease and are easy to relapse, which are difficult to be cured, seriously affecting people's life and health. NOD-like receptor pyrin domain-containing 3 (NLRP3) inflammasome is an important component of inflammatory response and innate immune system. The inflammatory cascade induced by NLRP3 activation is involved in the occurrence and development of DM as well as its complications by releasing inflammatory factors, damaging endothelial cells and affecting metabolic stress. Therefore, as the core of the inflammatory response, NLRP3 may provide a new target for the treatment of DM and its complications. Traditional Chinese medicine plays a key role in the treatment of DM and its complications, and has a regulatory effect on NLRP3. Thus it has become a novel research strategy to prevent and treat DM and its complications via modulating NLRP3. However, at present, there are relatively scattered reports and a lack of systematic review on the role of traditional Chinese medicine in the treatment of DM and its complications from the perspective of NLRP3. As a result, this paper reviewed domestic and foreign literature in recent years and conducted the discussion from two aspects: the influence of NLRP3 on the occurrence and development of DM and its complications, and the progress of traditional Chinese medicine in intervening in DM and its complications through NLRP3. This paper provided reference for the research on the regulation of NLRP3 and a new direction for the treatment of DM and its complications.

17.
Article in Chinese | WPRIM | ID: wpr-940761

ABSTRACT

ObjectiveTo investigate the effect of modified Taohe Chengqitang on NOD-like receptor protein 3 (NLRP3) inflammasome activation in rats with diabetic cardiomyopathy. MethodSPF male SD rats aged 3-4 weeks were randomly divided into a normal group and an experimental group. The rats in the experimental group were fed on a high-fat diet for 4 weeks and then received intraperitoneal injection of streptozotocin (STZ) at 35 mg·kg-1 to induce the diabetes model. The rats in the experimental group were randomly divided into model group, low- and high-dose modified Taohe Chengqitang groups (11.7 g·kg-1 and 23.4 g·kg-1), and metformin hydrochloride group (67.5 mg·kg-1) according to the fast blood glucose (FBG). The cardiac function and structure of rats were detected by ultrasonic imaging after 8 weeks of continuous intragastric administration. Blood samples from the femoral artery were collected to detect FBG, triglyceride (TC), and total cholesterol (TG) of rats. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in rat myocardium. Serum levels of interleukin-1β (IL-1β) and interleukin-18 (IL-18) were determined by enzyme-linked immunosorbent assay (ELISA). The protein expression of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cysteinyl aspartate-specific protease 1 (Caspase-1), and phosphorylated nuclear factor kappa-B p65 (p-NF-κB p65) in the myocardium was detected by Western blot. ResultCompared with the normal group, the model group showed increased levels of FBG, TC, and TG (P<0.01), decreased left ventricular ejection fraction (EF) and left ventricular fractional shortening (FS) (P<0.05), myocardial hypertrophy and myocardial fibrosis as revealed by HE staining, increased serum levels of 1L-1β and 1L-18 and protein expression of NLRP3, ASC, Caspase-1, and p-NF-κB p65 in myocardial tissues (P<0.01). Compared with the model group, the modified Taohe Chengqitang groups and the metformin group showed reduced levels of FBG, TC, and TG (P<0.05), restored EF and FS (P<0.05), improved pathological changes in myocardial tissues, and decreased serum levels of IL-1β and IL-18 and protein expression of NLRP3, ASC, Caspase-1, and p-NF-κB p65 in myocardial tissues (P<0.05). The improvement was more significant in the high-dose modified Taohe Chengqitang group (P<0.05). ConclusionModified Taohe Chengqitang can protect the myocardium by inhibiting the activation of NLRP3 inflammasomes.

18.
Article in Chinese | WPRIM | ID: wpr-940689

ABSTRACT

ObjectiveTo observe the effects of Sinisan on behaviors and NOD-like receptor protein 3 (NLRP3) inflammasomes of depressed rats induced by chronic unpredictable mild stress (CUMS) and further explore the anti-depressant mechanism of Sinisan. MethodFifty male rats were randomly divided into a normal group, a model group, an NLRP3 inhibitor (MCC950) group (10 mg·kg-1), and low- (2.5 g·kg-1) and high-dose (5 g·kg-1) Sinisan groups, with 10 rats in each group. The depression model was induced by 42 d CUMS in rats except for those in the normal group. Drug intervention was performed on the 22nd day of modeling by gavage in the Sinisan groups and by intraperitoneal injection in the MCC950 group. Except for the MCC950 group, the remaining four groups received 10 mg·kg-1 physiological saline by intraperitoneal injection, while the rats in the model group, the normal group, and the MCC950 group were administered with 3 mL of physiological saline by gavage. Twenty-one days later, the sucrose preference test and open field test were performed. Western blot was used to detect the protein expression of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cysteinyl aspartate-specific protease-1 (Caspase-1), B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), ionized calcium-binding adapter molecule 1 (Iba1), and CD68 in the hippocampus of rats in each group. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of interleukin-1β (IL-1β) and interleukin-18 (IL-18) in the hippocampus of rats. Nissl staining and TUNEL were used to assess the pathological changes and apoptosis level in the hippocampal CA1 region of rats, respectively. ResultThe sucrose preference rate and consumption volume in the sucrose preference test, the total distance, the percentage of central movement distance, and the percentage of residence time in the open field test of rats in the model group were lower than those in the normal group (P<0.01). Compared with the model group, the Sinisan groups and the MCC950 group showed improved depression-like behaviors, apoptosis level in the hippocampal CA1 region, and neuron loss to varying degrees. Sinisan could reduce the levels of IL-1β, IL-18, Bax, Iba1, and CD68 in the hippocampus (P<0.05, P<0.01), increase the level of Bcl-2 (P<0.05, P<0.01), and inhibit the protein expression of NLRP3, ASC, and Caspase-1 related to NLRP3 inflammasomes (P<0.05, P<0.01). ConclusionSinisan can improve the depression-like behaviors and pathological damage of hippocampal neurons in CUMS-induced rats, and the mechanism may be related to the inflammatory response mediated by the NLRP3 inflammasome signaling pathway.

19.
Article in Chinese | WPRIM | ID: wpr-940350

ABSTRACT

ObjectiveTo explore the possible mechanism of total flavonoids of peony flower (TFPF) in protecting rats from gouty nephropathy and provide data support for the pharmaceutical research on the treatment of gouty nephropathy. MethodGouty nephropathy rat model was established by adenine combined with ethambutol. Rats were randomly assigned into blank control group, model group, allopurinol (42 mg·kg-1) group, Tongfengshu tablets (600 mg·kg-1, positive control) group, and TFPF (260, 130, and 65 mg·kg-1) groups. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1), interleukin-1β (IL-1β), and interleukin-18 (IL-18) in rat serum and those of transforming growth factor-β1 (TGF-β1) and IL-1β in renal homogenate. Hematoxylin-eosin(HE) staining was carried out for observation of the morphological changes of renal cells. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) was conducted for observation of the DNA damage in renal cells. The expression of NOD-like receptor protein 3 (NLRP3), cysteine aspartic acid protease(Caspase)-1 and IL-1β were observed by immunohistochemistry. The expression levels of NLRP3, Caspase-1 and nuclear transcription factor -κB (NF-κB) in renal tissues were detected by Western blot. ResultCompared with blank group, the contents of TNF-α, MCP-1, IL-1β, IL-18, and TGF-β1 in serum of model group were significantly increased (P<0.01), and the expressions of NLRP3, Caspase-1, NF-κB and IL-1β in kidney of model group were significantly increased (P<0.01). The renal tissue cells showed cytoplasmic swelling, cell membrane rupture, and the number of nuclear pyknotic fracture increased. The positive rate of TUNEL staining was significantly increased in model group (P<0.01), and the contents of IL-1β and TGF-β1 in renal tissue homogenate were significantly increased (P<0.01). Compared with model group, the contents of inflammatory factors TNF-α, IL-1β and IL-18 in serum of rats in TFPF high- and medium-dose groups could be decreased to different degrees (P<0.05, P<0.01), while the content of MCP-1 in TFPF high-dose group was significantly decreased (P<0.01). The content of TGF-β1 in renal tissue homogenate in TFPF high- and medium-dose groups was significantly decreased (P<0.05, P<0.01), and the content of IL-1β in renal tissue homogenate in TFPF medium-dose group was significantly decreased (P<0.01). HE staining showed that each dose group of TFPF could improve the status of renal tubular epithelial cells, reduce cytoplasmic swelling and the number of nuclear pyknosis to varying degrees. The positive rate of TUNEL staining was decreased (P<0.01) and DNA damage was decreased. The expression of NLRP3, Caspase-1, IL-1β and NF-κB protein in renal tissue cells was inhibited (P<0.05, P<0.01). ConclusionTFPF protects rats from gouty nephropathy by inhibiting the secretion of inflammatory cytokines. Specifically, it may inhibit the activation of NF-κB and NLRP3/Caspase-1 pathways to reduce the expression, maturation, and release of inflammatory cytokines such as IL-1β and IL-18 and further inhibit pyroptosis, thereby reversing the inflammatory injury of kidney in gouty nephropathy.

20.
Article in Chinese | WPRIM | ID: wpr-940227

ABSTRACT

Ischemic stroke, also known as cerebral infarction, is the most common type of stroke. Ischemic stroke is extremely harmful with high rates of morbidity, incidence, disability, and mortality, bringing a huge burden on society and families. As a result, finding new and effective prevention and treatment methods is critical. The pathological mechanism of ischemic stroke is very complex and superimposed, with inflammatory response serving as a critical pathological link in the ischemic stroke cascade injury process. NOD-like receptor 3 (NLRP3) is an intracellular sensor, and the inflammatory cascade mediated by the activated NLRP3 inflammasome can exacerbate ischemic stroke injury through the release of inflammatory factors. Taking the NLRP3 inflammasome as the entry point, a large number of experimental studies on the intervention of traditional Chinese medicine (TCM) in the assembly and activation of NLRP3 inflammasome have been carried out, which proved that Chinese medicinal monomers or prescriptions with the main functions of tonifying deficiency, clearing heat and removing toxin, eliminating phlegm, promoting circulation and resolving stasis can interfere with the assembly and activation of NLRP3 inflammasome, reduce the inflammatory response, and relieve ischemic stroke. This study reviewed the assembly and activation of NLRP3 inflammasome, the mechanism of NLRP3 inflammasome in ischemic stroke, and the prevention and treatment of ischemic stroke by TCM through regulation of the NLRP3 inflammasome, which provides a new entry point for the pathological mechanism of ischemic stroke and a direction for the development of new treatments for ischemic stroke.

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