Papel del IFN-λ en la patogénesis del Lupus eritematoso sistémico: mecanismos Biomoleculares

Introducción: El lupus eritematoso sistémico (LES) es una enfermedad autoinmune que causa inflamación sistémica y alteraciones en la tolerancia inmunológica. La activación de los genes inducibles por interferón (IFN), contribuye en más del 50% de su patogenia. Objetivo: relacionar el papel del IFN-λ en la patogenia del LES. Materiales y Métodos: Búsqueda sistémica en base de datos; a través de las palabras claves del MeSH and DeCS. Fue incluido adicionalmente la palabra "Interferón Lambda". Resultados: Se encontró que la producción aberrante de interferón tipo I contribuye a la desregulación de IFN-λ, producido principalmente por células dendríticas plasmocitoides. Este proceso conduce a la estimulación inmunológica por autoanticuerpos y a un aumento de IFNλR-1 en células B, potenciando la generación de anticuerpos. IFN-λ3 se asocia particularmente con nefritis lúpica, y el IFN-λ en general aumenta la expresión de MHC-I, intensificando la respuesta de células T CD8+ y posiblemente afectando la tolerancia central y la regulación en el timo. Conclusión: Se destaca que el IFN-λ favorece la activación inmune, formación de inmunocomplejos, inflamación crónica y producción de autoanticuerpos, vinculándose niveles altos de IFN-λ3 con mayor actividad de la enfermedad.

Introduction: Systemic lupus erythematosus (SLE) is an autoimmune disease that causes systemic inflammation and alterations in immunological tolerance. The activation of interferon (IFN)-inducible genes contributes to more than 50% of its pathogenesis. Objective: to review the role of IFN-λ in the pathogenesis of SLE. Materials and Methods: Systemic search in database; through the MeSH and DeCS keywords. The word "Lambda Interferon" was additionally included. Results: Aberrant production of type I interferon was found to contribute to the deregulation of IFN-λ, produced mainly by plasmacytoid dendritic cells. This process leads to immunological stimulation by autoantibodies and an increase in IFNλR-1 in B cells, enhancing the generation of antibodies. IFN-λ3 is particularly associated with lupus nephritis, and IFN-λ generally increases MHC-I expression, enhancing the CD8+ T cell response and possibly affecting central tolerance and regulation in the thymus. Conclusion: It is highlighted that IFN-λ favors immune activation, formation of immune complexes, chronic inflammation and production of autoantibodies, linking high levels of IFN-λ3 with greater disease activity.

Effects of Bushen Huoxue Formula （补肾活血方） on TLR/NF-κB Pathway and Intestinal Flora in Ileum Tissue of Parkinson's Disease Model Mice / 中医杂志

ObjectiveTo explore the possible mechanism of Bushen Huoxue Formula （补肾活血方， BHF） in the treatment of Parkinson's disease （PD） from the the perspective of intestinal flora. MethodsSeventy-two male C57/BL6J mice were randomly divided into blank group， model group， Madopar group and low-， medium- and high-dose BHF groups， with 12 mice in each group. The mice in the blank group were intraperitoneally injected with 10 ml/kg of normal saline， and those in the other groups were intraperitoneally injected with 30 mg/kg of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine （MPTP） at a concentration of 3 mg/ml to induce PD mice model， both once a day for 7 consecutive days. After successful modeling， the low-， medium-， and high-dose BHF groups were given 7.5， 15， and 30 g/（kg·d） of BHF by gavage， respectively， while the Madopar group was given 112.5 mg/（kg ·d） of Domedopar tablets by gavage， and the blank group and the model group were given 15 ml/（kg·d） of distilled water， all once a day for 14 consecutive days. The rod climbing test， rotating rod test， grip strength test and weight-bearing swimming test were used to evaluate the behavioral indicators of mice. Western blotting was used to measure the protein expression levels of Toll-like receptor （TLR）/nuclear factor kappa B （NF-κB） pathway inflammatory factors in the mouse ileum， including Toll-like receptor 2 （TLR2）， Toll-like receptor 4 （TLR4）， NF-κB， tumor necrosis factor α （TNF-α）， interleukin 6 （IL-6）， and interleukin 17 （IL- 17）. 16S rRNA high-throughput sequencing was used to analyze changes in mouse intestinal flora. ResultsCompared to those in the blank group， the mice in the model group had longer bottoming time when climbing the pole， reduced grip strength， shortened rotary pole duration and swimming duration， and increased protein expression levels of TLR2， TLR4， NF-κB， TNF-α， and IL-6 in the ileal tissue （P＜0.01）. Compared to the model group， the Madopar group and the low-， medium- and high-dose BHF groups had shortened bottoming time of the climbing pole and increased grip strength； the Madopar group and the high-dose BHF group had prolonged rotary pole duration， and reduced protein expressions of TLR2， TLR4， NF-κB， TNF-α， IL-6， and IL-17 levels； and only the high-dose BHF group had prolonged swimming duration （P＜0.05 or P＜0.01）. Compared to those in the low-dose BHF group， the bottoming time of the climbing pole were shorter in the moderate- and high-dose groups （P＜0.05 or P＜0.01）， and the grip strength increased while the protein expression levels of TLR2， TLR4 and IL-17 decreased in the high-dose group （P＜0.05 or P＜0.01）. The intestinal flora results showed significant differences between the blank group and the model group in the Dominance index， Pielou_e index， Shannon index， and Simpson index （P＜0.05 or P＜0.01）. Compared to those of the model group， the Shannon index， Chao1 index， and Observed_otus index of the Madopar group， as well as the Chao1 index， Observed_otus index， Dominance index， Pielou_e index， Shannon index， and Simpson index of the high-dose BHF group all showed significantly statistical differences （P＜0.05 or P＜0.01）. At the phylum level， the relative abundance categories of bacterial phyla with statistically significant differences in each group included Proteobacteria， Bacteroidetes， and Firmicutes （P＜0.05 or P＜0.01）. At the genus level， the relative abundance categories of bacterial genera with statistically significant diffe-rences among each group included Muribaculaceae， Akkermansia， and Helicobacter pylori （P＜0.05 or P＜0.01）. ConclusionThe possible mechanism of BHF in treating PD may be to reconstruct the disordered intestinal flora structure and improve the inflammatory response.

The relationship between the expression of TLR2 and TLR4 in peripheral blood mononuclear cells and Th1/Th2 immune response in children with recurrent respiratory tract infection / 国际检验医学杂志

Objective To investigate the expression of Toll-like receptor 2(TLR2)and Toll-like receptor 4(TLR4)in peripheral blood mononuclear cells(PBMC)in children with recurrent respiratory tract infection(RRTI)and its relationship with T helper cell 1(Th1)/T helper cell 2(Th2)immune response.Methods A total of 65 children diagnosed with RRTI who admitted to the hospital from December 2020 to December 2022 were enrolled in the study as the RRTI group,and 45 healthy children who underwent physical examination in the hospital during the same period were enrolled as the control group.The relative expression levels of TLR2 and TLR4 mRNA in PBMCs were detected by real-time fluorescence quantitative PCR(qPCR).The expres-sion rates of TLR2 and TLR4 protein in PBMCs were detected by flow cytometry.The levels of Th1 cytokine interferon-γ(IFN-γ),Th2 cytokine interleukin-4(IL-4)and their ratio(IFN-γ/IL-4)in plasma were detected by enzyme-linked immunosorbent assay(ELISA).Pearson correlation analysis was used to analyze the corre-lation between TLR2,TLR4 protein expression rates and plasma IFN-γ,IL-4 levels.Results The RRTI group had significantly higher plasma level of Th2 cytokine IL-4 than the control group,significantly lower plasma level of Th1 cytokine IFN-y than the control group,and significantly lower ratio of IFN-γ/IL-4 than the con-trol group,the differences were all statistically significant(P＜0.05).The relative expression levels of TLR2 and TLR4 mRNA and protein expression rates in PBMC of children in the RRTI group were higher than those in the control group,and the differences were statistically significant(P＜0.05).Pearson correlation analysis showed that the protein expression rates of TLR2 and TLR4 in PBMC of children with RRTI were both nega-tively correlated with both plasma IFN-γ levels and IFN-γ/IL-4(P＜0.05)and positively correlated with plasma IL-4 levels(P＜0.05).Conclusion The expression of TLR2 and TLR4 in PBMC and plasma Th1/Th2 cytokines in children with RRTI may be involved in the occurrence and development of the disease.Ex-cessive activation of TLR2 and TLR4 may weaken Th1 function and enhance Th2 function.

Expression of Toll like receptor in pancreatic cancer cells / 中华肝胆外科杂志

Objective:To compare the expression difference of Toll like receptor (TLR) and inflammatory factors between pancreatic cancer and normal pancreatic epithelial cells, and explore the correlation between TLR and inflammatory microenvironment.Methods:Normal pancreatic duct epithelium cells (HPNE) and pancreatic cancer cells (Panc-1 and Mia-PACA-2) were cultured and proteins were obtained. The expression of TLR family protein, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and myeloid differentiation factor 88 (MyD88) were examined by western blot in HPNE, Panc-1 and Mia-PACA-2. The correlations between TLR and inflammation cytokines of pancreatic cancer were analyzed by Pearson correlation analysis.Results:Compared with HPNE, the TLR2, TLR3, TLR4, TLR7, TLR8 and TLR9 were highly expressed in Panc-1 and Mia-PACA-2 (all P<0.05). Compared with Panc-1, the expression of TLR2 and TLR4 in Mia-PACA-2 were increased obviously, while the TLR9 expression was mildly decreased (all P<0.05). The expression of IL-6 in HPNE was found less than that in Panc-1 (0.52±0.03 vs. 0.76±0.04) and Mia-PACA-2 (0.52±0.03 vs. 1.12±0.09) with statistical differences ( P<0.05). Similarly, the expression of TNF-α was found significantly less than that of Panc-1 cells (0.63±0.04 vs. 0.87±0.06) and Mia-PACA-2 cells (0.63±0.04 vs. 0.95±0.10) with statistical differences (all P<0.05). The expression of IL-6 was found positively correlated with expressions of TLR2 ( r=0.964), TLR4 ( r=0.968), TLR7 ( r=0.844), TLR8 ( r=0.668) (all P<0.05), and the expression of TNF-α was found positively correlated with expressions of TLR2 ( r=0.805), TLR4 ( r=0.893), TLR7 ( r=0.847), TLR8 ( r=0.780) (all P<0.05). In contrast with HPNE, the expression of MyD88 was found highly expressed in Panc-1 (0.91±0.10 vs. 0.33±0.03) and Mia-PACA-2 (1.14±0.10 vs. 0.33±0.03) (all P<0.001). Compared with Panc-1, the expression of MyD88 in Mia-PACA-2 was obviously increased (1.14±0.10 vs. 0.91±0.10) with statistical difference ( P=0.048). Conclusion:The TLR family may play a critical role in development of pancreatic cancer by regulating the immune microenvironment, and its mechanism may be through upregulating MyD88 which functions as key signal transduction.

The effects and mechanisms of Toll like receptors 4/9 mediating IL-10, IL-35 and TGF-β expressions by CD25+B cells from periodontitis individuals / 西安交通大学学报(医学版)

【Objective】 To analyze the expressions of IL-10, IL-35 and TGF-β in CD25+B cells from periodontitis individuals, and then establish how the activation of TLR4/9 affects the above processes. 【Methods】 SD rats were randomly divided into healthy group, primary periodontitis groups and severe periodontitis group; experimental models were performed by ligation. Expression of IL-10, IL-35 and TGF-β mRNA in CD25+B cells from gingiva and peripheral blood, expression and activation of TLR 2/4/7/9, MyD88, TRAF6 in gingival CD25+B cells were detected. The effect of TLRs/MyD88 on IL-10, IL-35 and TGF-β expressions and production were evaluated by cell culture experiments. 【Results】 CD25+B cells from gingiva of primary periodontitis individuals showed improved expression of IL-10 and TGF-β mRNA compared with the healthy ones (P<0.05); cells from peripheral blood did not present the same tendency. CD25+B cells from gingiva of severe periodontitis individuals showed improved expression of IL-10, IL-35 and TGF-β mRNA compared with the healthy ones (P<0.05), cells from peripheral blood showed higher IL-10 mRNA level than the healthy ones (P<0.05). Compared with healthy individuals, the expression and phosphorylation of TLR4/9 and MyD88 in CD25+B cells from gingiva of severe periodontitis individuals were increased (P<0.01). In cell culture experiments, TLR4 agonist promoted IL-10, IL-35 and TGF-β mRNA expression and IL-10 secretion (P<0.05); TLR9 agonist improved IL-10 and TGF-β mRNA expression and IL-10 secretion (P<0.05). The combined use of TLR4/9 agonist could increase the expression and secretion of all the detected indexes (P<0.05); MyD88 antagonism decrease the above effects (P<0.05). 【Conclusion】 The expressions of IL-10, IL-35 and TGF-β in gingiva CD25+B cells increase during periodontitis, which may be regulated by TLR4 /9-MyD88 pathway.

Research on expression of Toll-like receptors and IFN-α in HCV infected patients with different clinical outcomes / 中国热带医学

@#Abstract: Objective To investigate the changes in expression of Toll-like receptor 3 (TLR3) and interferon-α (IFN-α) in patients with different clinical outcomes of hepatitis C virus (HCV) infection treated with direct-acting antiviral agents (DAAs), and to explore the relationship between the expression of TLR3 and IFN-α with the clinical outcomes of HCV infection. Methods A total of 149 HCV infected patients who received initial treatment were selected from Hainan General Hospital between September 2020 and August 2022. The patients were divided into two groups: chronic hepatitis C (CHC) group (n=129) and liver cirrhosis (LC) group (n=20). Additionally, 28 volunteers were selected as the control group during the same period. All patients with HCV infection were first treated with Sofosbuvir/Vipatavir tablets for 12 weeks. Blood samples were collected at 0, 4, 12, 24 and 48 weeks after treatment. Liver function indicators were detected by enzyme-linked immunosorbent assay (ELISA), while TLR3 mRNA were detected by real-time fluorescence quantitative polymerase chain reaction (qRCR), IFN-α was detected by Luminex multiplex cytokine assays. Measurement data subject to normal distribution were represented by x±s, and t test was used between groups. Compare differences between groups. Results TLR3 mRNA in CHC group was higher than that in LC group and control group at baseline (P<0.05). After 4 weeks of DAAs treatment, TLR3 mRNA in CHC and LC groups was significantly up-regulated (P<0.05). TLR3 mRNA in the CHC group was gradually down-regulated to the level of the control group at 12, 24, and 48 weeks. In addition, IFN-α expression gradually increased with prolonged treatment, while it decreased in the LC group. The liver inflammation indicators in both the CHC and LC groups partially recovered after treatment with DAAs. Conclusions TLR3 is involved in viral clearance and chronic inflammatory response. The expression difference of TLR3 in patients with different clinical outcomes of HCV infection after DAAs treatment may be related to the severity of the disease.

Research progress of Toll-like receptors and pulmonary infection / 中国医师杂志

Due to the increase of susceptible individual species and drug-resistant pathogens, pulmonary infection is still a global problem that threatens human health. Toll-like receptors (TLR) can identify pathogenic pattern related molecules on the surface of pathogenic microorganisms in innate immunity and initiate innate immune response. It can participate in different anti-infection mechanisms and play an important role in pulmonary infectious diseases caused by different pathogens. In this paper, the research progress of TLR in pulmonary infection at home and abroad in recent years is reviewed.

Study on the peripheral blood levels changes of soluble receptor for advanced glycosylation end product, endotoxin and Toll-like receptor in patients with severe acute pancreatitis complicated with peritoneal cavity infection / 中国医师进修杂志

Objective:To explore the peripheral blood levels changes of soluble glycosylation end product receptor (sRAGE), endotoxin and Toll-like receptor (TLR) in patients with severe acute pancreatitis (SAP) complicated with peritoneal cavity infection, and clarify the relationship between indexes and pathogenetic condition.Methods:The clinical data of 105 patients with SAP in Shanghai Pudong New Area People′s Hospital from January 2017 to December 2020 were retrospectively analyzed. Among them, 28 cases had peritoneal cavity infection (infection group), and 77 cases had peritoneal cavity infection symptoms but undiagnosed (non-infection group). The acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) at admission was recorded. When patients had the suspected symptoms and signs of peritoneal cavity infection, the serum levels of sRAGE, endotoxin, TLR4 and TLR9 were detected by enzyme-linked immunosorbent assay. The correlation between serum levels of sRAGE, endotoxin, TLR-4, TLR-9 and APACHE Ⅱ was analyzed by Pearson analysis; the risk factors of peritoneal cavity infection in SAP patients were analyzed by multivariate Logistic regression; the receiver operating characteristic (ROC) curve was drawn, and the diagnostic efficacy of serum sRAGE, endotoxin, TLR-4 and TLR-9 in peritoneal cavity infection were evaluated in patients with SAP; the Kaplan-Meier survival curve was drawn, and the comparison used log-rank test.Results:The serum sRAGE, endotoxin, TLR-4 and TLR-9 in infection group were significantly higher than those in non-infection group: (822.16 ± 104.51) ng/L vs. (728.09 ± 96.47) ng/L, (62.59 ± 20.11) ng/L vs. (41.62 ± 13.64) ng/L, (45.17 ± 8.54) μg/L vs. (37.34 ± 6.22) μg/L, (26.35 ± 6.73) μg/L vs. (20.02 ± 5.49) μg/L, and there were statistical differences ( P<0.01). Pearson correlation analysis result showed that the serum sRAGE, endotoxin, TLR-4 and TLR-9 were positively correlated with APACHE Ⅱ in patients with SAP ( r = 0.632, 0.556, 0.521 and 0.631; P<0.05). Multivariate Logistic regression analysis result showed that the combined organ function damage, shock, hypoxemia and serum sRAGE, endotoxin, TLR-4 and TLR-9 were independent risk factors of peritoneal cavity infection in patients with SAP ( P<0.05). ROC curve analysis result showed that the area under the curve for the serum sRAGE, endotoxin, TLR-4 and TLR-9 combined diagnosis of peritoneal cavity infection in patients with SAP was the largest, which was 0.910 (95% CI 0.838 to 0.957, P<0.01), with a sensitivity of 82.14% and a specificity of 87.01%. According to the ROC curve cut-off value of serum sRAGE, endotoxin, TLR-4 and TLR-9 (764.58 ng/L, 58.01 ng/L, 40.24 μg/L and 22.61 μg/L), the 28 patients with SAP complicated with peritoneal cavity infection were divided into high levels patients (21, 14, 23 and 22 cases) and low levels patients (7, 14, 5 and 6 cases); Kaplan-Meier survival curve analysis result showed that the 28-day survival rates in patients with high levels of sRAGE, endotoxin, TLR-4 and TLR-9 were significantly lower than those in patients with low levels (61.90% vs. 71.43%, 50.00% vs. 78.57%, 60.87% vs. 80.00% and 59.09% vs. 83.33%), and there were statistical differences ( P<0.05). Conclusions:The serum sRAGE, endotoxin, TLR-4 and TLR-9 have a high combined diagnostic value in SAP complicated with peritoneal cavity infection, and they are all related to the severity of the disease and have a significant impact on survival.

Anti-tumor and Phenotypic Regulation Effect of Matrine on Dendritic Cells through Regulating TLRs Pathway / 中国结合医学杂志

OBJECTIVE@#To investigate the effects of matrine on antigen presentation of dendritic cells (DCs), and to explore the pharmacological mechanism of matrine on anti-tumor effect.@*METHODS@#Different concentrations (0, 1, 2, 4, 8 and 16 µ g/mL) of matrine were co-cultured with DCs, the harvested DCs were co-cultured with antigens of Lewis lung cancer (LLC) cells, and then DCs and T cells were co-cultured to produce DCs-activated killer (DAK) cells, which have significant tumor-killing activity. The expression of cytokines, mRNA and protein of toll-like receptors (TLRs) in DCs were detected by enzyme linked immunosobent assay, polymerase chain reaction and Western blot, respectively. And the killing effect of DAK were measured by MTT assay.@*RESULTS@#Matrine significantly increased the mRNA expression of TLR7, TLR8, myeloid differentiation factor 88 (MyD88), tumor necrosis factor receptor-associated factor 6 (TRAF-6) and I κ B kinase (IKK), as well as the protein expression of TLR7 and TLR8, and up-regulated the levels of interleukin-12 (IL-12), IL-6 and tumor necrosis factor-α (TNF-α), meanwhile, it also increased the expressions of MHC-II, CD54, CD80 and CD86 in DCs. DCs-activated effector T cells had significant tumor-killing activity. When the concentration of matrine was more than 4 µg/mL, all indices had significant difference (P<0.01 or P<0.05).@*CONCLUSION@#Matrine plays an anti-tumor role by regulating TLRs signal transduction pathway, promoting the secretion of inflammatory cytokines and enhancing immune function.

Association Between TLR2, TLR4 and NOD2/CARD15 Gene Polymorphisms and Ulcerative Colitis: A Meta-analysis / 胃肠病学

Background: The prevalence rate of ulcerative colitis (UC) in China has increased significantly in recent years, and Toll-like receptor 2 (TLR2), TLR4 and NOD2/CARD15 may be closely related to the development of UC. Aims: To explore the influence of TLR2, TLR4 and NOD2/CARD15 gene polymorphisms on the pathogenesis of UC. Methods: Studies on correlation of TLR2, TLR4 and NOD2/CARD15 gene polymorphisms with UC were retrieved from PubMed, CBM, CNKI, Wanfang, VIP databases. Literatures were enrolled according to the inclusion and exclusion criteria, and the quality was evaluated and data were extracted. RevMan 5.3 software was used to conducted meta-analysis. Results: Fifteen eligible articles were included. Meta-analysis showed that TLR2 Arg753Gln gene polymorphism was not associated with risk of UC (P>0.05). Except the recessive model, TLR4 Asp299Gly gene polymorphism could increase the risk of UC (P0.05). No significant association between NOD2/CARD15 (Arg702Trp, Gly908Arg and Leu1007fsinsC) gene polymorphism and UC was found (P>0.05). Conclusions: Existing evidence shows that TLR4 (Asp299Gly, Thr399Ile) gene polymorphisms are associated with an increased risk of UC, however, NOD2/CARD15 (Arg702Trp, Gly908Arg and Leu1007fsinsC) and TLR2 (Arg753Gln) gene polymorphisms are not associated with UC.

Hidroxicloroquina: Del fármaco de herencia Inca hasta la inmunología moderna / Hydroxychloroquine: Inca inheritance drug to modern immunology

Resumen La hidroxicloroquina es un antiguo fármaco proveniente del árbol de quino (Cinchona pubescens), a partir del componente químico alcaloide llamado quinina. Sus primeros usos se documentaron en el Imperio Inca del Perú. Se caracteriza por distintos efectos beneficiosos en enfermedades inmunológicas, al disminuir los procesos de autoinflamación y autoinmunidad persistente. Esta revisión se enfoca en describir los mecanismos inmunomoduladores de la hidroxicloroquina, así como los efectos del fármaco en algunas de las enfermedades autoinmunes más prevalentes: lupus eritematoso sistémico, artritis reumatoide, síndrome de Sjögren, vasculitis sistémicas, nefropatía por IgA, síndrome antifosfolípido, distintas enfermedades inmunológicas de la piel. También se revisarán los efectos adversos descritos para este fármaco, especialmente la toxicidad de retina, que es el más temido.

Abstract Hydroxychloroquine is an old drug derived from the quino tree (Cinchona pubescens), from the alkaloid chemical component called quinine. Its first uses trace back to the Inca empire of Peru. It is characterized by different beneficial effects in immunological diseases, decreasing the processes of autoinflammation and persistent autoimmunity. This review focuses on describing the immunomodulatory mechanisms of hydroxychloroquine as well as the effects of the drug on some of the most prevalent autoimmune diseases: systemic lupus erythematosus, rheumatoid arthritis, Sjögren's syndrome, systemic vasculitis, IgA nephropathy, antiphospholipid syndrome, different skin-related autoimmune disorders. The main adverse effects will be revised, focusing in the retinal toxicity.

Effect of Coptidis Rhizoma-Zingiberis Rhizoma Extract on Colonic Epithelium TLR4/NF-κB Signaling Pathway in Mice with Ulcerative Colitis / 中国实验方剂学杂志

Objective::To observe the effect of berberine and 6-shogaol, main components of Coptiae Rhizoma and Zingiberis Rhizoma, on the inflammatory signaling pathway of Toll-like receptors 4 (TLR4)/nuclear factor kappa B (NF-κB) in colonic epithelial cells of mice with ulcerative colitis. Method::Fifty Kunming mice were randomly divided into normal group, model group, berberine group (100 mg·kg-1), 6-shogaol group (100 mg·kg-1), and 6-shogaol combined with berberine group (200 mg·kg-1), with 10 mice in each group. A mouse model of ulcerative colitis was established through oral administration with 2% dextroan sulfate for two weeks. Each group was given corresponding drugs by gavage, while normal group and model group were given equal amount of normal saline. Serum and colon tissue samples were taken 20 days after administration. Enzyme-linked immunosorbent method was used to detect serum interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) expressions, and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot method were used to detect TLR4, NF-κB p65 mRNA and protein expressions in colon epithelial tissue. Result::Compared with the normal group, relative expressions of TLR4 and NF-κB p65 mRNA and protein were increased in the model group (P<0.01), and the contents of serum IL-1β and TNF-α were increased (P<0.01). Compared with the model group, relative expressions of TLR4 and NF-κB p65 mRNA and protein were significantly decreased in 6-shogaol group, berberine group and 6-shogaol combined with berberine group (P<0.01), and the contents of serum IL-1β and TNF-α were significantly decreased (P<0.01). Among the three groups, 6-shogaol combined with berberine group had the strongest effect (P<0.01). Conclusion::Both 6-shogaol and berberine can inhibit colonic inflammation, reduce inflammatory damage and treat ulcerative colitis. The combined application of 6-shogaol and berberine has a significant synergism effect. The mechanism is related to the excessive activation of TLR4/NF-κB pathway and the regulation of non-controllable intestinal inflammation.

Insights into immunoregulatory properties of dental-derived mesenchymal stem cells in oral diseases / 中国组织工程研究

BACKGROUND: Mesenchymal stem cells have strong immunoregulatory capabilities. The immunoregulatory characteristics of dental-derived mesenchymal stem cells are closely related to the occurrence and development of oral diseases. However, the specific mechanism of this immunomodulatory effect on the disease is unknown. Therefore, the application of the immunomodulatory properties of dental derived mesenchymal stem cells may play an important role in the treatment and prevention of oral diseases in the future. OBJECTIVE: To review the role of the Immunomodulatory properties of dental-derived mesenchymal stem cells in oral diseases from the three following aspects: the mutual regulation mechanism of mesenchymal stem cells derived from different oral tissues and activated immune cells, and the effects of various cytokines and oral pathogens on the immunoregulatory capacity of dental-derived mesenchymal stem cells. METHODS: CNKI and PubMed databases were retrieved for relevant literature published from January 2000 to June 2019. The search terms were “tooth-derived mesenchymal stem cells, immunomodulatory properties, immune cells, oral diseases, Porphyromonas gingivalis, periodontitis, dental plaque, dental caries, toll-like receptors” in Chinese and English, respectively. A total of 221 articles were obtained, 64 of which met the standards for result analysis. RESULTS AND CONCLUSION: In the past 20 years since the dental-derived mesenchymal stem cells were isolated, researchers have conducted a large number of in vitro experiments on the interaction between several oral tissue-derived mesenchymal stem cells and activated immune cells. The mechanism of action between them has been preliminarily discussed. Different inflammatory mediators and cytokines can differentially activate various immunomodulatory proteins in mesenchymal stem cells, and thus activate their immunomodulatory activities. Therefore, exploration on specific stimuli that activate mesenchymal stem cells and oral pathogenic bacteria to activate dental-derived mesenchymal stem cells may improve the application efficiency of dental-derived mesenchymal stem cells at different stages of the tissue healing process. It will make a breakthrough in the way that the disease is treated.

Interferon-gamma combined with lipopolysaccharide polarizes human umbilical cord-derived mesenchymal stem cells to a MSC2 phenotype / 中国组织工程研究

BACKGROUND: Mesenchymal stem cells have been widely applied in autoimmune diseases and graft-versus-host diseases because of their immunomodulatory capabilities. However, mesenchymal stem cells have plasticity in immunomodulation, which leads to heterogeneity and instability when used in vivo. OBJECTIVE: To investigate the polarization of human umbilical cord derived mesenchymal stem cells to an immunosuppressive phenotype (MSC2) in the inflammatory microenvironment induced by interferon-γ and lipopolysaccharide. METHODS: Human umbilical cord-derived mesenchymal stem cells were isolated and cultured in vitro, and then were identified by morphological characteristics, surface markers, adipogenesis and osteoinduction activity. Human umbilical cord-derived mesenchymal stem cells were treated with interferon-γ (10 μg/L), lipopolysaccharide (100 μg/L), or their combination for 24 hours, respectively, and were then co-cultured with phytohemagglutinin pre-treated peripheral blood mononuclear cells for 5 days under direct or Transwell indirect contact. The percentages of regulatory T cells and T helper 1 cells were analyzed by flow cytometry at different times. The mRNA expression levels of Toll-like receptors 2, 3 and 4 were detected by real-time fluorescence quantitative PCR. RESULTS AND CONCLUSION: (1) Human umbilical cord derived mesenchymal stem cells exhibited spindle-shaped or fibroblast-like morphology, highly expressed CD73, CD90 and CD105, and lacked expression of CD34, CD45 and HLA-DR. (2) Under direct or indirect co-culture, human umbilical cord-derived mesenchymal stem cells pre-treated with interferon-γ and lipopolysaccharide could promote the generation of regulatory T cells, which was superior to the interferon-γ, lipopolysaccharid, un-treated and control groups (P < 0.05). (3) The percentage of T helper 1 cells gradually decreased over time. (4) Under indirect co-culture, human umbilical cord derived mesenchymal stem cells pre-treated with interferon-γ and lipopolysaccharide were polarized into immunosuppressive MSC2 phenotype at an earlier period and highly expressed Toll-like receptor 3 (P < 0.05). (5) In conclusion, the combination of interferon-γ (10 μg/L) and lipopolysaccharide (100 μg/L) results in the high-efficient polarization of mesenchymal stem cells toward the MSC2 phenotype under indirect co-culture, and the immunosuppressive capability of MSC2 is independent of intercellular contact, which provides clinical evidence for the MSC2-derived exosome therapy in the future.

Roles of Platelet Toll-like Receptors in Thrombosis / 中国医学科学院学报

Platelets are non-nuclear blood cells that are widely involved in physiological and pathological processes.Their main role is to participate in hemostasis and thrombosis.Toll-like receptors(TLRs)are innate immune receptors.Platelets express multiple TLRs and can promote thrombosis by recognizing ligand-induced platelet activation and aggregation.This article reviews the relationship between platelets/TLR and thrombosis and the roles of TLRs in the development of thrombotic diseases.

Role of hepatic macrophages in the development and progression of primary biliary cholangitis / 临床肝胆病杂志

Primary biliary cholangitis(PBC) is a chronic autoimmune liver disease characterized by cholestasis and has unknown etiology. Its specific pathogenesis remains unclear, but hepatic macrophages are one of the key aspects of the immune and inflammatory response of PBC and the injury of biliary epithelial cells. Hepatic macrophages are classified into resident Kupffer cells and monocyte-derived macrophages according to their source, and these cells play an important role in the development and progression of PBC. This article reviews the role of hepatic macrophages in the development and progression of PBC.

Progress in Research on Correlation Between Toll-like Receptors and Barrett's Esophagus / 胃肠病学

Barrett's esophagus (BE), the only recognizable precancerous lesion of esophageal adenocarcinoma (EAC), is a common disease characterized by abnormal transformation of squamous epithelium into intestinal columnar epithelium. Toll-like receptors (TLRs) are important pattern recognition receptors that play a central role in innate and adaptive immunity. TLRs are involved in the identification of almost all pathogens. In addition, TLRs signaling pathway is important for the occurrence and development of BE. This article reviewed the progress in research on the correlation between TLRs and BE.

Regulatory T cells isolated from endometriotic peritoneal fluid express a different number of Toll-like receptors / Células T reguladoras isoladas do fluido peritoneal de mulheres com endometriose expressam um número diferente de receptores do tipo Toll

ABSTRACT Objective To analyze and compare the expression of Toll-like receptors by regulatory T cells present in the peritoneal fluid of patients with and without endometriosis. Methods Regulatory T cells were isolated from peritoneal fluid of women with and without endometriosis, collected during surgery, and mRNA was extracted for analysis of Toll-like receptors expression by reverse-transcriptase polymerase chain reaction. Results Patients with endometriosis presented regulatory T cells expressing a larger number and variety of Toll-like receptors when compared to regulatory T cells from patients in the Control Group. Toll-like receptor-1 and Toll-like receptor-2 in regulatory T cells were expressed in both groups. All other expressed Toll-like receptors types were only found in regulatory T cells from the Endometriosis Group. Conclusion Patients with endometriosis had peritoneal regulatory T cells expressing various Toll-like receptors types.

RESUMO Objetivo Analisar e comparar a expressão de receptores do tipo Toll por células T reguladoras presentes no líquido peritoneal de pacientes com endometriose. Métodos Células T reguladoras foram isoladas do líquido peritoneal de mulheres com e sem endometriose, coletadas durante a cirurgia, e o RNAm foi extraído para análise da expressão de receptores do tipo Toll por reação em cadeia da polimerase com transcriptase reversa. Resultados Pacientes com endometriose apresentaram células T reguladoras expressando maior número e variedade de Toll por células quando comparadas com T reguladoras de pacientes do Grupo Controle. Receptores do tipo Toll-1 e receptores do tipo Toll-2 foram expressos em ambos os grupos. Todos os outros tipos de receptores Toll foram encontrados expressos apenas em células T reguladoras do grupo com endometriose. Conclusão Pacientes com endometriose apresentaram células T reguladoras peritoneais expressando vários tipos de receptores tipo Toll.

Pam2CSK4 (TLR2 agonist) induces periodontal destruction in mice

Abstract Lipoproteins are important bacterial immunostimulating molecules capable of inducing receptor activator of nuclear factor-κB (RANKL) and osteoclast formation in vitro and in vivo . Although these molecules are present in periodontopathogenic bacteria, their role in periodontitis is not known. In this study, we used Pam2CSK4 (PAM2), a synthetic molecule that mimics bacterial lipoprotein, to investigate the effects of lipoproteins on periodontitis in mice. C57BL/6 male mice were randomly divided into three experimental groups: 1) Negative control group: animals received vehicle injection; 2) Positive control group: animals received injection of Escherichia coli lipopolysaccharide (LPS); 3) PAM2 group: animals received PAM2 injection. All the injections were performed bilaterally every other day into the palatal mucosa between first and second molars. After twenty-four days, the animals were euthanized to assess alveolar bone volume (micro-CT), cellular and extracellular composition in the gingiva (stereometric analysis), and osteoclast numbers (TRAP staining). Treatment with either PAM2 or LPS induced gingival inflammation, as demonstrated by increased infiltration of inflammatory cells and enhanced angiogenesis, associated with a smaller number of fibroblasts and decreased extracellular matrix. Importantly, treatment not only with LPS but also with PAM2 resulted in a larger number of TRAP+ multinucleated osteoclasts and significant loss of alveolar bone. Collectively, our data demonstrate that PAM2 can induce gingival inflammation and bone loss in mice, broadening the avenues of investigation into the role of lipoproteins in the pathogenesis of periodontal disease.

Pam2CSK4 (TLR2 agonist) induces periodontal destruction in mice

Abstract Lipoproteins are important bacterial immunostimulating molecules capable of inducing receptor activator of nuclear factor-κB (RANKL) and osteoclast formation in vitro and in vivo . Although these molecules are present in periodontopathogenic bacteria, their role in periodontitis is not known. In this study, we used Pam2CSK4 (PAM2), a synthetic molecule that mimics bacterial lipoprotein, to investigate the effects of lipoproteins on periodontitis in mice. C57BL/6 male mice were randomly divided into three experimental groups: 1) Negative control group: animals received vehicle injection; 2) Positive control group: animals received injection of Escherichia coli lipopolysaccharide (LPS); 3) PAM2 group: animals received PAM2 injection. All the injections were performed bilaterally every other day into the palatal mucosa between first and second molars. After twenty-four days, the animals were euthanized to assess alveolar bone volume (micro-CT), cellular and extracellular composition in the gingiva (stereometric analysis), and osteoclast numbers (TRAP staining). Treatment with either PAM2 or LPS induced gingival inflammation, as demonstrated by increased infiltration of inflammatory cells and enhanced angiogenesis, associated with a smaller number of fibroblasts and decreased extracellular matrix. Importantly, treatment not only with LPS but also with PAM2 resulted in a larger number of TRAP+ multinucleated osteoclasts and significant loss of alveolar bone. Collectively, our data demonstrate that PAM2 can induce gingival inflammation and bone loss in mice, broadening the avenues of investigation into the role of lipoproteins in the pathogenesis of periodontal disease.