Human intestinal flora refers to a large and diverse microbial population present in the digestive tract of the human body, which plays a significant role in the establishment of human immune homeostasis and the normal function of the immune system. Myasthenia Gravis is an autoimmune disease of the neuromuscular junction, mainly involved in the anti-acetylcholine receptor antibody, cellular immune dependence, and complement1. At present, studies have found that the intestinal flora of Myasthenia Gravis is different from that of healthy people. Probiotic therapy has been shown effective in the experimental autoimmune Myasthenia Gravis animal models. This article reviews the relationship between intestinal flora and Myasthenia Gravis, to provide new ideas for further study of the pathogenesis and clinical treatment of Myasthenia Gravis.
Objective: Based on bioinformatics, gene set enrichment analysis (GSEA) and immune infiltration analysis were carried out on the microarray data of psoriasis expression profile to further understand the pathogenesis of psoriasis. Methods: GSE6710 chip data were obtained from gene expression database (GEO), and gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were performed using GSEA software. 22 kinds of immune cell gene expression matrices and R packages were downloaded from CIBERSOFT official website, and the immune cell infiltration matrix was obtained by R software and related graphs were drawn. Results: The pathways related to cell proliferation and innate immunity were highly expressed in psoriatic lesions, and some cancer-related pathways were highly expressed in psoriatic lesions. Immunized cell infiltration analysis showed that activated memory T cells, follicular helper T cells, M0 macrophages and activated dendritic cells were up-regulated in psoriatic skin lesion group, and inactive mast cells were down-regulated in psoriatic skin lesion group. Activated dendritic cells are positively correlated with follicular helper T cells, activated mast cells are positively correlated with M0 macrophages. Inactivated mast cells are negatively correlated with activated memory T cells, M1 macrophages are negatively correlated with regulatory T cells, M0 macrophages are negatively correlated with inactive mast cells. Conclusion: Cell proliferation and innate immunity are of great significance in the pathogenesis of psoriasis. Immune cell infiltration analysis is generally consistent with the current psoriasis pathogenesis model. Macrophages and mast cells also play a certain role in psoriasis.
Objective:To establish an experimental model of chronic obstructive pulmonary disease(COPD)in rats,and explore whether polyadenosine diphosphate ribose polymerase-1(PARP-1)inhibitors regulate Sirtuin 1(SIRT1)and peroxisome proliferator-activated receptor-γ co-activator 1α(PGC-1α)to reduce inflammation and oxidative stress in COPD rats,and explore the possibility of SIRT1-PGC-1α axis as a new target of PARP-1 inhibitor.Methods:Twelve of 48 SD rats were randomly selected as healthy group,and the remaining rats were used to construct experimental models of COPD.Rats that were successfully modeled were randomly divided into model group,PARP-1 inhibitor treatment group and PARP-1 inhibitor+PGC-1α inhibitor group.HE staining was used to observe pathological changes of lung tissue,ELISA was used to detect levels of TNF-α,IL-6,IL-1β,malondialdehyde(MDA),superoxide dismutase(SOD)in rats lung tissue,fluorescence quantitative PCR was used to detect expression levels of SIRT1 and PGC-1α mRNA of rats in each group,Western blot was used to detect expressions of SIRT1 and PGC-1α proteins.Results:Lung tissue structure of rats in healthy group was complete,compared with healthy group,lung tissue of model group suffered structural damage,with a large number of inflammatory cells infiltrated,contents of TNF-α,IL-1β and IL-6 in alveolar lavage fluid were significantly increased,con-tent of MDA in serum was significantly increased,while content of SOD was significantly reduced,and expressions of SIRT1,PGC-1α mRNA and protein were significantly reduced;compared with model group,lung tissue structure of rats in PARP-1 inhibitor treatment group was recovered,and inflammatory cells were reduced,contents of TNF-α,IL-1β and IL-6 were significantly reduced,content of MDA in serum was significantly reduced,while content of SOD was significantly increased,and expressions of SIRT1,PGC-1α mRNA and protein were significantly increased;compared with PARP-1 inhibitor treatment group,the number of inflammatory cells in PARP-1 inhibitor+PGC-1α inhibitor group was increased,contents of TNF-α,IL-1β and IL-6 were significantly increased,content of MDA in serum was significantly increased,while content of SOD was significantly reduced,expressions of SIRT1,PGC-1α mRNA and protein were significantly reduced.Conclusion:PARP-1 inhibitors can alleviate inflammation and oxidative stress by activating SIRT1-PGC-1α axis,thereby effectively alleviating COPD.
Objective:To explore the mechanism of dendritic cells(DCs),novel regulatory B cells(B10 cells)and Th17/Treg imbalance in the pathogenesis of patients with chronic obstructive pulmonary disease(COPD)and their correlation with lung function.Methods:According to the"Guidelines for the Diagnosis and Treatment of Chronic Obstructive Pulmonary Disease"a total of 93 COPD patients were prospectively selected from the Ninth People's Hospital of Suzhou from May 2019 to December 2021,and 50 healthy subjects were selected as the control group.The patients were followed up for 1 year to observe the occurrence of acute exacer-bation COPD(AECOPD),and divided them into stable COPD group and AECOPD group.The course of disease,modified British Medical Research Society dyspnea index(mMRC)classification,COPD assessment test(CAT)score,BODE index score,6 min walking distance(6MWD),arterial partial pressure of oxygen(PaO2),arterial carbon dioxide Partial pressure(PaCO2)were com-pared between the two groups;compared the levels of FEV1,FVC,FEV1/FVC,and the percentage of FEV1 to predicted value(FEV1/Pred)in the three groups with peripheral blood DCs,B10 cells,Th17 cells,Treg cells and Th17/Treg,IL-12,IL-10,IL-17A and TGF-β1 levels.To analyze the correlation between peripheral blood DCs cells,B10 cells and Th17/Treg imbalance and pulmonary function indexes in AECOPD group.Logistic regression analysis of independent risk factors for AECOPD.Results:A total of COPD pa-tients had AECOPD events(40.86%).The course of disease,mMRC grade,CAT score,BODE index score,and PaCO2 in AECOPD group were significantly higher than those in COPD stable group(P<0.05),6MWD and PaO2 were significantly lower than those in COPD group.The levels of FEV1,FVC,FEV1/FVC and FEV1/Pred in the AECOPD group were significantly lower than those in the stable COPD group and control group(P<0.05);the levels of FEV1,FVC,FEV1/FVC and FEV1/Pred in the stable COPD group were significantly lower than those in control group(P<0.05).DCs,B10 cells and Treg cells in AECOPD group were significantly lower than those in stable COPD group and control group,while Th17 expression level and Th17/Treg were significantly higher than that in stable COPD group and control group(P<0.05).DCs,B10 cells and Treg cells in stable COPD phase were significantly lower than those in control group,while Th17 expression level and Th17/Treg were significantly higher than control group(P<0.05).The ex-pression levels of IL-12,IL-10 and TGF-β1 in the AECOPD group were significantly lower than those in the stable COPD group and control group(P<0.05),while IL-17A was significantly higher than that in the stable COPD group and control group.The expression levels of IL-12,IL-10 and TGF-β1 in patients with stable COPD were significantly lower than control group,while IL-17A was signifi-cantly higher than control group(P<0.05).Pearson analysis showed that peripheral blood DCs,B10 cells were positively correlated with FEV1,FVC,FEV1/FVC and FEV1/Pred levels(P<0.05),while Th17/Treg was positively correlated with FEV1,FVC,FEV1/FVC and FEV1/Pred levels all were negatively correlated(P<0.05).Logistic regression analysis found that mMRC grade and Th17/Treg were independent risk factors of AECOPD(P<0.05).Conclusion:With the progression of COPD,DCs,B10 cells,Th17 cells,Treg cells and Th17/Treg gradually become unbalanced,resulting in disordered expression levels of pro-inflammatory and anti-inflamma-tory factors.Peripheral blood DCs and B10 cells were positively correlated with lung function levels,while Th17/Treg were negatively correlated with lung function levels.mMRC grade and Th17/Treg are independent risk factors of AECOPD.Therefore,actively inter-vening in the imbalanced state of immune function in patients has specific and important clinical significance in reducing the immune damage of lung tissue and promoting the improvement of lung function.
As the main weapon of cellular immunity,CD4+ T cells play a vital role in controlling and eliminating infections,and are an important barrier for the body to resist infections.Respiratory tract infectious diseases caused by influenza virus infection have extremely high infectivity,morbidity and mortality.The infection mechanism is relatively complicated and has not been fully ex-plained.The exuberant immune response induced by the body after influenza virus infection is described as a"cytokine storm"which is related to pro-inflammatory cytokines and tissue damage,which may eventually lead to acute lung injury.Therefore,this article sum-marizes the current research progress,focusing on the mechanism of CD4+T cells in the cytokine storm induced by influenza virus in-fection and the impact of acute lung injury,providing relevant ideas and theoretical guidance for follow-up research,with a view to the disease caused by influenza virus bring new and effective methods of diagnosis and treatment.
Objective:To preliminarily screen the key genes of primary immune thrombocytopenia(ITP)by bioinformatics method and explore the pathogenesis,so as to predict the potential traditional Chinese medicine(TCM)for the treatment of ITP.Methods:Based on the original microarray data set GSE80401 under the National Center for Biotechnology Information(NCBI),the differential miRNA of ITP were obtained by analyzing the adjusted P<0.05 and |logFC|≥1 as the screening criteria for differential miRNA.miRTarBase,miRDB and TargetScan were used to predict miRNA target genes.The target of ITP was searched in Genecards database,and the predicted up-regulated target genes and down-regulated target genes were intersected with disease targets.On this basis,the mapped target genes were respectively constructed into PPI network through String database and Cytoscape to screen core target genes,and the core target genes were enriched and analyzed in DAVID and Omicsbean databases for GO and KEGG pathways.The key genes were imported into the Coremine Medical database to analyze the TCM for the treatment of key genes.Results:Total of 422 differential genes and 17 key genes were finally screened,including BCL2L1,CCND1,CD44,CDKN1A,CREB1,GRB2,MAPK1,MAPK8,PIK3R1,CDK2,CAV1,FGF2,IGF1,SMAD2,SMAD4,TLR4 and VEGFA,mainly involving proteoglycan,FoXO,PI3K-Akt,human T cell leukemia virus 1 infection,endocrine resistance,focal adhesion and other signal pathways.A total of 12 TCM for ITP prevention and treatment,including ginseng,Radix Paeoniae Rubra,Angelica sinensis,bee venom,cobra,Psoralea corylifolia,Rehmannia glutinosa,buffalo horn,hemp seed,dodder seed,Wulingzhi and Jinji NaPi were screened.TLR4 maps the most TCM,followed by CCND1 and VEGFA.Among many TCM,ginseng acts on 17 targets at the same time,Radix Paeoniae Rubra,Angelica sinensis and bee venom act on 11 targets at the same time,cobra and Psoralea corylifolia act on 9 and 8 targets at the same time,Rehmannia glutinosa and buffalo horn act on 7 targets at the same time,hemp seed act on 4 targets at the same time,dodder seed act on 3 targets at the same time,and wulingzhi act on 2 targets at the same time.It is suggested that these drugs had the potential of multi-target prevention and treatment of ITP.Conclusion:The key pathogenic genes of ITP and the TCM with preventive and thera-peutic effects could be preliminarily predicted based on the analysis of genetic information,which can provide targets and research ideas for the development of related TCM.
Objective:To investigate the effect of astragaloside Ⅳ on the apoptosis of thyroid cells in Hashimoto's thyroiditis(HT)rats and Ras homolog gene family member A(RhoA)/Rho-associated coiled-coil containing kinase 2(ROCK2)pathway.Methods:The HT rat model was induced by subcutaneous injection of thyroglobulin combined with high iodine drinking water and randomly divided into model group,astragaloside(80 mg/kg)group,Rhosin(RhoA inhibitor,40 mg/kg)group,astragaloside Ⅳ(80 mg/kg)+ Rhosin(40 mg/kg)group(12 rats in each group),another 12 SD rats were selected and drank water normally and injected the same dose of saline subcutaneously as control group.After the drugs were grouped and processed,the serum anti-thyroglobulin antibody(TGAb),anti-thyroid peroxidase antibody(TPOAb)levels and the inflammatory factors IL-6,IL-17,IL-1β contents were measured by ELISA kits;the pathological changes of thyroid tissue in each group were detected by hematoxylin-eosin(HE)staining;the apopto-sis rate of rat thyroid cells in each group were detected by TUNEL staining;the expressions of RhoA/ROCK2 pathway proteins in thy-roid tissues of rats in each group were detected by Western blot.Results:Compared with the control group,the thyroid follicles in the model group had abnormal structure,some atrophy or disappearance,disordered arrangement,surrounding inflammatory cell infiltra-tion,and obvious pathological damage to the thyroid tissue,the serum TGAb,TPOAb,IL-6,IL-17 and IL-1β levels,thyroid cell apoptosis rate,and thyroid tissue RhoA and ROCK2 protein expression levels were significantly increased(P<0.05);compared with model group,the pathological damage of the thyroid tissue of rats in the drug intervention group were reduced,the serum TGAb,TPOAb,IL-6,IL-17 and IL-1β levels,thyroid cell apoptosis rate,and thyroid tissue RhoA and ROCK2 protein expression levels were decreased(P<0.05);compared with astragaloside Ⅳ group and the Rhosin group respectively,the pathological damage of the thyroid tissue of rats in the astragaloside Ⅳ+Rhosin group were further reduced,the serum TGAb,TPOAb,IL-6,IL-17 and IL-1β levels,thyroid cell apoptosis rate,thyroid tissue RhoA and ROCK2 protein expression levels were decreased(P<0.05).Conclusion:Astragaloside Ⅳ may down-regulate the expression of RhoA/ROCK2 pathway to reduce the inflammatory injury of thyroid tissue,inhib-it thyroid cell apoptosis,and improve the symptoms of HT in rats.
Objective:To investigate the regulation of melatonin(MT)against lung injury in a mouse model of influenza virus infection by regulating the maternally expressed gene 3(MEG3)/microRNA-223(miR-223)/nucleotide-binding oligomerization do-main-like receptor protein 3(NLRP3)axis.Methods:Fifty mice were randomly divided into control group,model group,and MT(low,medium,and high)dose groups,with 10 mice in each group.Except for the control group,the other groups were given H7N9 virus suspension nasal drops at a dose of 5×105 EID50,and the control group was given an equal volume of phosphate buffer nasal drops.The MT(low,medium,and high)dose groups were injected intraperitoneally with(15,30,60)mg/kg MT on the day after modeling,the control group and model group were intraperitoneally injected with equal volume of normal saline.The mice were sacri-ficed 24 h after the last administration for experiment.All mice were tested for the lung index;Hematoxylin-Eosin(HE)staining was used to observe the lung tissue morphology;ELISA was used to detect the levels of interleukin IL-1β,IL-18,IL-6,TNF-α and IFN-β in lung tissues;real-time fluorescent quantitative PCR(RT-qPCR)was used to detect the levels of MEG3 mRNA and miR-223 in mouse lung tissues;Western blot was used to detect the protein levels of NLRP3,apoptosis-related dot-like protein(ASC),cysteine aspartate proteolytic enzyme 1(caspase-1),and pro-caspase-1 in lung tissues.Bioinformatics prediction and dual luciferase experi-ments were used to detect the targeting relationship between MEG3 and miR-223,miR-223 and NLRP3.Results:The pathological re-sults showed that the alveolar wall of the lung tissue of the model group was obviously congested,and there was obvious inflammatory exudation and inflammatory cell infiltration in the cavity;with the increase of the MT dose,the hyperemia of the alveolar wall of the mouse lung tissue,the inflammatory exudation in the cavity and the infiltration of inflammatory cells in the MT(low,medium,and high)dose group were all improved.Compared with control group,the lung index,chemokines,inflammatory factors,antiviral fac-tors,MEG3 levels in lung tissues,NLRP3 pathway protein level in the model group were increased(P<0.05),and the miR-223 level was decreased(P<0.05);after the addition of MT,the antiviral factors increased significantly,and other indicators were improved.miR-223 had targeted regulatory relationships with MEG3 and NLRP3.Conclusion:MT can alleviate the lung injury of H7N9 influen-za virus-infected mice,which may be related to the regulation of MEG3/miR-223/NLRP3 axis to relieve inflammation.
Objective:To explore the correlation between differential immune-related genes(DIRGs)and immune cells in the progression of atherosclerosis(AS)and general rule of Chinese medicine for intervening DIRGs.Methods:Firstly,GSE28829 data set was obtained from GEO database,and immune-related genes were downloaded from ImmPort database and MSigDB database.Secondly,differentially expressed genes between early AS plaques(EAP)and advanced AS plaques(AAP)of GSE28829 were screened by limma package,and their intersections with immune-related genes were known as DIRGs.clusterProfiler package was used to enrich the DIRGs.Protein interaction network of DIRGs was constructed and Hub genes were screened.Then,the infiltration patterns of 22 kinds of immune cells were analyzed by CIBERSORT to screen differential immune cells,and the correlation between them and Hub genes were analyzed by Pearson method.Finally,Coremine Medical database was used to predict Chinese herbs for in-tervening DIRGs,the property and flavor of Chinese herbs were collected.Results:Total 63 DIRGs were obtained,and 10 Hub genes(CD86,TLR2,TYROBP,CCR1,ITGB2,CCL2,CCL4,CSF1R,CXCR4,CTSS)were screened out.Enrichment analysis results showed that the molecular functions,biological processes and signaling pathways of DIRGs were closely related to immune regulation.Analysis of immune cell infiltration showed that proportions of regulatory T cells,activated dendritic cells and resting mast cells in EAP were increased.Proportions of memory B cells,γδ T cells,M0 macrophages and M2 macrophages were increased in AAP.Correla-tion analysis showed that CD86 was positively correlated with M2 macrophages in EAP.In AAP,CD86,CTSS,CXCR4,CSF1R,ITGB2 and TYROBP were positively correlated with M0 macrophages,while CCL4 and CCL2 were negatively correlated with resting mast cells.Results of frequency showed that Chinese herbs for intervening DIRGs mainly distributed to liver and lung meridians,and their property and taste were cold and bitter.Conclusion:This study found that in the progression of AS,10 DIRGs were the most important,and 7 kinds of immune cells were dysregulated,among which CD86,CTSS,CXCR4,CSF1R,ITGB2,TYROBP,CCL4 and CCL2 were correlated with resting mast cells,M0 and M1 macrophages.At the same time,immune mechanism of AS progression was closely related to liver meridian,lung meridian,bitter,sweet,cold and warm.The results can provide reference and ideas for Chinese herbs clinical prescription to treat AS and further exploratory the immunological mechanism of AS progression.
Objective:To investigate the effect and mechanisms of mitochondrial dynamic-related protein 1(Drp1)on the in-flammatory response after intracerebral hemorrhage(ICH)in mice.Methods:Western blot was used to detect the expressions of Drp1 and phosphorylated Drp1(p-Drp1)in brain tissues after ICH,and the optimal concentration of selective Drp1 inhibitor(Mdivi-1)were screened.The mice were randomly divided into sham group,ICH group,ICH+solvent control group(ICH+Vehicle group)and ICH+Mdivi-1 group.The neurological function was evaluated by mNSS,the content of brain water was measured by wet-dry weight method,HE and Nissl staining were used to observe the morphological changes of brain tissues,neutrophil infiltration were observed by MPO staining,and the expression of inflammatory cytokines(IL-6,TNF-α)and mitochondrial membrane marker proteins TOM20,COX4Ⅰ1 were detected by Western blot.Results:Compared with sham group,the expression of p-Drp1 was significantly increased at 12 h after ICH(P<0.01).Compared with ICH+Vehicle group,the neurological function score(P<0.01)and brain water content(P<0.05)of ICH+Mdivi-1 group were increased,the protein expressions of IL-6 and TNF-α and the number of MPO positive cells around hematoma in ICH+Mdivi-1 group were significantly increased(P<0.05),and the expressions of mitochondrial membrane proteins TOM20 and COX4Ⅰ1 in ICH+Mdivi-1 group were significantly elevated(P<0.01).Conclusion:Inhibition of Drp1 can inhibit mito-chondrial division and aggravate inflammatory injury after ICH in mice.Therefore,Drp1 may reduce the inflammation after ICH.
Objective:To explore the mechanism of Tanshinone ⅡA(TⅡA)in improving ischemia/reperfusion(I/R)injury of H9c2 cardiomyocytes by activating Sirtuin 1(SIRT1)-adenosine 5'-monophosphateactivated protein kinase(AMPK)pathway through miR-155-5p.Methods:H9c2 cells were cultured in vitro and I/R damage model was established.After modeling,H9c2 cells were randomly divided into model group,TⅡA group,TⅡA+miR-NC group,TⅡA+miR-155-5p mimics group,10 µmol/L TⅡA was added for intervention after transfection,and the H9c2 cells supplemented with DMSO were used as control group.qRT-PCR was used to detect expression level of miR-155-5p;MTT method was used to analyze cell proliferation ability;flow cytometry was used to evaluate cell apoptosis;ELISA was used to determine the levels of TNF-α,IL-4,IL-10,IL-17,lactate dehydrogenase(LDH),malo-ndialdehyde(MDA)and superoxide dismutase(SOD);Western blot was used to detect relative expressions of SIRT1,AMPK and p-AMPK proteins.Results:Compared with control group,expression of miR-155-5p in model group was increased,cell viability was decreased,apoptosis rate and expressions of TNF-α,IL-17,LDH and MDA were increased,while expressions of IL-4,IL-10,SOD,SIRT1 and p-AMPK were decreased(P<0.05);compared with model group,expression of miR-155-5p in TⅡA group was reduced,cell viability was increased,apoptosis rate and expressions of TNF-α,IL-17,LDH and MDA were decreased,while expressions of IL-4,IL-10,SOD,SIRT1 and p-AMPK were increased(P<0.05);compared with TⅡA group and TⅡA+miR-NC group,expression of miR-155-5p in TⅡA+miR-155-5p mimics group was increased,cell viability was decreased,apoptosis rate and expressions of TNF-α,IL-17,LDH and MDA were increased,while expressions of IL-4,IL-10,SOD,SIRT1 and p-AMPK were decreased(P<0.05).Conclusion:TⅡA can improve I/R injury of H9c2 cardiomyocytes by down-regulating miR-155-5p,and its mechanism may be related to the activation of SIRT1-AMPK pathway.
Objective:To investigate effects of extract of pinellia(EP)on proliferation and apoptosis of airway smooth muscle cells(ASMCs)in asthmatic rats and its mechanism.Methods:Asthma rat model was constructed by sensitization and challenge with ovalbumin and rat ASMCs were isolated and cultured.Immunofluorescence staining was used to identify α-actin in ASMCs.After iden-tifying as meeting characteristics of ASMCs,ASMCs were divided into control group,model group,EP group,Compound C group and EP+Compound C group.MTT was used to detect cell proliferation activity;flow cytometry was used to detect cell apoptosis;ELISA was used to detect levels of inflammatory factors IL-6 and TNF-α in cell supernatant;Western blot was used to detect expressions of CyclinD1,proliferating cell nuclear antigen(PCNA),Bcl-2 associated X protein(Bax),Caspase-3,Cleaved-Caspase-3,adenylate activated protein kinase(AMPK),p-AMPK,forkhead box protein O3a(FOXO3a),p-FOXO3a proteins.Results:Immunofluores-cence staining showed that 98%of cells showed green fluorescent filaments in cytoplasm,and α-actin was positively expressed,which proved that cultured cells were ASMCs.Compared with control group,cell OD490,CyclinD1,PCNA protein expressions and IL-6 and TNF-α levels in supernatant of model group were significantly increased,apoptosis rate,Bax,Caspase-3,Cleaved-Caspase-3 protein expressions and p-AMPK/AMPK,p-FOXO3a/FOXO3a levels were significantly reduced(P<0.05);compared with model group,cell OD490,CyclinD1,PCNA protein expressions and IL-6,TNF-α levels in supernatant of EP group were significantly reduced,apoptosis rate,Bax,Caspase-3,Cleaved-Caspase-3 protein expressions,and p-AMPK/AMPK,p-FOXO3a/FOXO3a levels were significantly increased(P<0.05),the above corresponding indicators of Compound C group showed opposite trend(P<0.05);compared with EP group,cell OD490,CyclinD1,PCNA protein expressions and IL-6,TNF-α levels in supernatant of EP+Compound C group were signifi-cantly increased,apoptosis rate,Bax,Caspase-3,Cleaved-Caspase-3 protein expressions,and p-AMPK/AMPK,p-FOXO3a/FOXO3a levels were significantly reduced(P<0.05).Conclusion:EP may inhibit proliferation of ASMCs and promote cell apoptosis in asthmatic rats by activating AMPK/FOXO3a pathway.
Objective:To analyze the difference of expression of Wnt protein family member 5a(Wnt5a)in prediabetes and diabetes mellitus and its correlation with immune inflammation in patients with prediabetes.Methods:From November 2020 to Novem-ber 2021,64 patients with prediabetes who treated in Zaozhuang Municipal Hospital were selected as group A,51 patients with diabe-tes were selected as group B,and 50 healthy people who underwent physical examination were selected as group C.The baseline data of 3 groups were collected,level of Wnt5a was determined,expression difference of Wnt5a in prediabetes and diabetes was analyzed,the related indicators of immune inflammation were determined,and the correlation between Wnt5a and immune inflammation in patients with prediabetes was analyzed.Predictive value of Wnt5a for prediabetes and diabetes was analyzed by ROC curve,and its cutoff values were determined for clinical disease prediction.Results:Compared with group B,Wnt5a level,immune inflammatory indexes Th1,Th17,Th22,IFN-γ,IL-17A and IL-22 levels in group A were decreased(P<0.05).Logistic regression analysis showed that fasting blood glucose,Wnt5a,Th1,Th17,Th22,IFN-γ,IL-17A and IL-22 were independent risk factors for the occurrence of prediabetes(P<0.05).Wnt5a was positively correlated with immune inflammation-related indicators Th1,Th17,Th22,IFN-γ,IL-17A and IL-22 in patients with prediabetes(P<0.05).ROC curve showed that AUC values of Wnt5a in predicting prediabetes and diabetes were 0.921 and 0.897,respectively,and the AUC were all>0.7,which indicating that Wnt5a had a higher predictive value for predia-betes and diabetes(P<0.05).Conclusion:There is a certain difference in expression of Wnt5a in prediabetes and diabetes.When level of 42.38 pg/ml≤Wnt5a<62.50 pg/ml,it is prediabetes,and when level of Wnt5a≥62.50 pg/ml,it indicates that the transition from prediabetes to diabetes has occurred,and believed that Wnt5a is related to immune inflammation in prediabetic patients,which may increase the degree of insulin resistance by amplifying local immune inflammation in prediabetic patients and eventually lead to the occurrence of diabetes.
Objective:To investigate whether etomidate affects inflammatory response and apoptosis of PC12 cells induced by hypoxia by regulating miR-142-3p.Methods:PC12 cells were pretreated with different doses(2,6,12 µmol/L)of etomidate to establish hypoxia model;PC12 cells that transfected with miR-142-3p mimics or inhibitors were pretreated with 0 or 12 µmol/L of etomidate to establish hypoxia model.Cell viability,apoptosis and protein(CyclinD1,Cleaved-caspase-3)expressions were detected by CCK-8 method,flow cytometry and Western blot,respectively.ELISA was used to detect levels of inflammatory factors TNF-α,IL-1β,IL-6.Expression of miR-142-3p was detected by RT-qPCR.Results:Etomidate increased hypoxia-induced PC12 cells activity and expres-sion of CyclinD1 protein and miR-142-3p,while decreased cell apoptosis rate,Cleaved-caspase-3 protein expression and levels of inflammatory factors TNF-α,IL-1β,IL-6(P<0.05).Up-regulation of miR-142-3p increased activity and expression of CyclinD1 pro-tein of hypoxia-induced PC12 cells,while decreased cell apoptosis rate,Cleaved-caspase-3 protein expression and levels of inflamma-tory factors TNF-α,IL-1β,IL-6(P<0.05).Down-regulation of miR-142-3p reversed effects of etomidate on hypoxia-induced PC12 cell activity,apoptosis and expressions of inflammatory factors(P<0.05).Conclusion:Etomidate can reduce inflammatory response and apoptosis of PC12 cells induced by hypoxia,and its mechanism may be related to the up-regulation of miR-142-3p expression in cells.
Objective:To investigate influences of ginsenoside Rg1 regulating miR-144-3p on neuroinflammation and blood-brain barrier damage in rats with experimental cerebral hemorrhage,and its regulation on formyl peptide receptor 2(FPR2)/p38 path-way.Methods:Ninety SD rats were randomly divided into control group,cerebral hemorrhage group,ginsenoside Rg1 low-dose group(10 mg/kg),ginsenoside Rg1 high-dose group(40 mg/kg),ginsenoside Rg1 high-dose+ago-miR-144-3p group(40 mg/kg ginseno-side Rg1+ago-miR-144-3p),with 18 mice in each group.Except for control group,experimental intracerebral hemorrhage rat model was constructed by injecting collagenase Ⅱ into right caudate nucleus,and then each group was given intraperitoneal administration and intracerebral injection as required.Neurological damage in rats was scored;rat brain water content was determined by dry-wet spe-cific gravity method;levels of TNF-α,IL-6 and IL-1β in rat brain tissues homogenate were determined by ELISA;ultrastructure around cerebral edema was observed by electron microscope;permeability of blood-brain barrier in rats was determined by Evans blue(EB)method;expressions of miR-144-3p/FPR2/p38 pathway were determined by qRT-PCR and Western blot.Results:Compared with control group,blood-brain barrier damage was aggravated in cerebral hemorrhage group,neurological function damage score,brain water content,miR-144-3p,TNF-α,IL-6,IL-1β,p38 mRNA,p-p38/p38 expressions in brain homogenate were increased(P<0.05),FPR2 mRNA and protein expressions were decreased(P<0.05);compared with cerebral hemorrhage group,blood-brain barrier damage was reduced in ginsenoside Rg1 low-dose group and ginsenoside Rg1 high-dose group,neurological function damage score,brain water content,miR-144-3p,TNF-α,IL-6,IL-1β,p38 mRNA,p-p38/p38 expressions in brain homogenate were decreased(P<0.05),FPR2 mRNA and protein expressions were increased(P<0.05);ago-miR-144-3p was able to reverse protective effects of gin-senoside Rg1 on blood-brain barrier and neuroinflammation in rats(P<0.05).Conclusion:Ginsenoside Rg1 may inhibit blood-brain barrier damage and neuroinflammation in rats by regulating miR-144-3p/FPR2/p38 axis.
Objective:To investigate effect and mechanism of mangiferin on regulation of M2-type macrophage polarization tar-geting MMP9 in pancreatic cancer.Methods:In vivo,therapeutic effect of mangiferin on pancreatic cancer was evaluated by drawing tumor growth curves and immunohistochemical staining.M2-type macrophages expression in pancreatic cancer was detected by immu-nofluorescence and ELISA.Effects of mangiferin on expression of MMP9 and downstream M2 macrophage polarization-related signaling pathways were detected by immunofluorescence,ELISA,Western blot and qRT-PCR.In vitro,MTT assay was utilized to detect effect of mangiferin on M2-type macrophage and therapeutic effect of mangiferin on pancreatic cancer.ELISA was used to detect effect of mangiferin on M2-type polarized macrophages.Effects of mangiferin on expression of MMP9 and its downstream signalling pathway were detected by immunofluorescence and Western blot.Results:Mangiferin had potential to inhibit growth of pancreatic cancer in mice pancreatic model,and could prevent expression of M2-polarized macrophages in pancreatic cancer in addition.At the same time,mangiferin could inhibit expression of MMP9 and downstream M2 macrophage polarization related signaling pathways in pancreatic cancer.Mangiferin inhibited proliferation of pancreatic cancer cells in a M2 type polarized macrophage-pancreas cancer cell co-culture model,inhibited macrophage M2 polarization,at the same time,expression of MMP9 and downstream M2 macrophage polarization related signaling pathway was inhibited.Conclusion:Mangiferin can inhibit macrophage M2 polarization by inhibiting MMP9 and its downstream signaling pathway,and play a role in pancreatic cancer therapy.
Objective:To explore the influence and possible mechanism of compound Huangbai liquid on wound healing in rats with skin ulcer.Methods:A rat model of skin ulcer was constructed and randomly separated into model group,compound Huang-bai liquid low-dose,medium-dose and high-dose groups(compound Huangbai liquid L,M,H groups)and erythromycin ointment group,10 rats/group;another 10 rats were randomly collected as the normal group.Compound Huangbai liquid L,M,H groups were given 2.5 ml,5 ml and 10 ml of compound Huangbai liquid,respectively;the erythromycin ointment group was given erythromycin ointment to spread evenly on the wound surface using cotton swabs;the model group and normal group were not given drug treatment.At 0 d,3 d,7 d and 14 d of administration,the wound healing of the rats were recorded by taking pictures,and the wound healing rate were calculated.On the 14th day of administration,the pathological changes and collagen deposition in the wounds of rats were determined by HE and Masson staining methods;the levels of IL-1β,TNF-α,nitric oxide(NO),transforming growth factor-β(TGF-β)and hydroxyproline in wound tissue were determined by kits;and the expressions of Notch1 and Twist1 proteins in wound tissue were determined by Western blot.Results:On the 7th and 14th day of administration,compared with model group,the wound healing rate of the rats in the compound Huangbai liquid L,M,H groups and the erythromycin ointment group were significantly improved(P<0.05),and the inflammatory cell infiltration and other phenomena in rats were significantly improved on 14 days of administration,and the deposition of collagen in the wound surface of rats was also significantly increased.On the 14th day of administration,com-pared with the normal group,the levels of IL-1β,TNF-α,NO,hydroxyproline and TGF-β in the wound tissues of the model group were significantly increased(P<0.05),the protein expressions of Notch1 and Twist1 were significantly decreased(P<0.05);com-pared with model group,the levels of IL-1β,TNF-α and NO in the compound Huangbai liquid L,M,H groups and erythromycin oint-ment group were significantly decreased(P<0.05),the levels of hydroxyproline and TGF-β and the protein expressions of Notch1 and Twist1 were significantly increased(P<0.05).Conclusion:Compound Huangbai liquid can promote wound healing in rats with skin ulcers by activating the Notch1/Twist1 axis.
Extracellular vesicles(EVs)are membrane-derived lipid bilayers secreted by bacteria and eukaryotic cells.EVs se-creted by Gram-negative bacteria have been extensively studied for many years,but due to differences in cell wall structure,EVs in Gram-positive bacteria have been disregarded.However,the production of EVs in Gram-positive bacteria was confirmed in recent stud-ies.Components of EVs are diverse,including proteins,lipids,nucleic acids and metabolites,play important role in biofilm forma-tion,antibiotic resistance,horizontal gene transfer(HGT)and pathogenesis.EVs have potential applications and are involved in the fields of vaccine research and development,disease treatment and drug delivery.Here,we review the latest findings on the biogene-sis,components,applications and functions of EVs from Gram-positive bacteria,hoping to provide a reference for the research in the field of EVs in Gram-positive bacteria.
Type 2 diabetes mellitus(T2DM)is a common chronic metabolic disease,while there is no consistent conclusion on the pathogenesis of T2DM.With the innovation of microecological research technology,intestinal dysbiosis is considered to be one of the causes of T2DM.Exercise is one of the effective means of diabetes management,while mechanism of regulating glucose metabo-lism still has many problems to be solved.In recent years,people have found that exercise can improve the intestinal environment and intestinal flora.This article will review the research on the influence and mechanism of exercise on T2DM based on intestinal flora,aiming to provide new ideas for exploring how exercise can improve T2DM in the future.
Objective:To investigate the neuroprotective effect of synthetic triterpenoid(CDDO-Im)on ischemic stroke rats and its influence on inflammatory response by activating the nuclear factor 2-related factor 2(Nrf2)/antioxidant response elements(ARE)signaling pathway.Methods:SD rats were divided into sham group(S group),MCAO group(M group)and CDDO-Im inter-vention group(M+C group).The middle cerebral artery occlusion(MCAO)was used in M group and M+C group to establish ischemic stroke model in rats,and sham operation was used in S group to control.After operation,M+C group was injected with CDDO-Im(64 µg/300 g)every12 hours via caudal vein,S group and M group were given the same amount of normal saline.After 3 days,the nerve function of rats was measured by Longa score,and the area of cerebral infarction was evaluated by 2,3,5-triphenyltetrazolium chlo-ride(TTC)staining.The expressions of Nrf2,heme oxygenated-1(HO-1),ionic calcium binding adaptor molecule 1(Iba1),IL-1β and IL-4 protein were detected by Western blot.Results:Compared with S group,M group rats showed significant neurologic deficit and cerebral infarction area,and the expression of Nrf2 protein had no significant difference,and the expression levels of HO-1,Iba1,IL-1β and IL-4 protein were increased significantly(P<0.05).Compared with M group,the neurological deficit score,cerebral infarction area,the expression levels of Iba1 and IL-1β protein were decreased significantly(P<0.05),while Nrf2,HO-1 and IL-4 pro-tein expression increased significantly in M+C group(P<0.05).Conclusion:CDDO-Im may activate the Nrf2/ARE signaling pathway and play a neuroprotective role,which may be related to the modulation of microglia to M2 and the regulation of inflammatory response.
