Correlation between tumor necrosis factor-α and interleukin-1β in exhaled breath condensate and pulmonary function in chronic obstructive pulmonary disease / 中国热带医学

@#Abstract: Objective To explore the correlation between lung function in patients with chronic obstructive pulmonary disease (COPD) and tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) levels in exhaled breath condensate (EBC), and to provide a convenient methodological basis for the diagnosis and treatment of COPD and the determination of its efficacy. Methods A total of 81 COPD patients and 40 healthy controls were selected from the respiratory department of the Fourth Affiliated Hospital of Guangzhou Medical University from August 2020 to February 2022 as the research subjects. The COPD patients were divided into 41 cases in the acute exacerbation group and 40 cases in the remission group according to their status. All participants underwent lung function detection, venous blood and EBC collection, and the levels of TNF-α and IL-1β in EBC and venous blood were analyzed by enzyme-linked immunosorbent assay (ELISA), and correlation analysis was performed by Pearson correlation analysis method. Results The levels of TNF-α and IL-1β in EBC of in the acute exacerbation group, the healthy control group, the remission group were (5.16±0.18) pg/μL and (7.75±0.27) pg/μL, (2.66±0.31) pg/μL and (2.41±0.24） pg/μL, (3.61±0.29) pg/μL and (3.17±0.38) pg/μL, respectively. Compared with the healthy control group, the levels of TNF-α and IL-1β in EBC in the COPD acute exacerbation group were significantly higher than those in the healthy control group and the COPD remission group (F=9.451, 8.217, P<0.001). Serum tests were consistent with this result. Correlation analysis showed that the levels of TNF-α and IL-1β in EBC were significantly positively correlated with the level of serum inflammation levels (P<0.001), while significantly negatively correlated with lung function (P<0.001).  Conclusions TNF-α and IL-1β in EBC are potential biomarkers of inflammation in patients with COPD, and their detection can be used to effectively assess lung function in patients with COPD. 

Effect of aloperine on chondrocyte injury induced by interleukin-1β and its mechanism / 中国生物制品学杂志

@#Objective To investigate the effect of aloperine(ALO)on interleukin-1β(IL-1β)-induced chondrocyte injury and its mechanism. Methods Chondrocytes were randomly divided into control(Con)group,IL-1 β group,IL-1β + ALO-L(25 mg/L)group,IL-1β + ALO-M(50 mg/L)group and IL-1 β + ALO-H(100 mg/L)group;Con group,IL-1βgroup,IL-1β + miR-NC group and IL-1β + miR-16-5p group;Con group,IL-1β group,IL-1β + si-NC group and IL-1β + siSOX5 group. Cells in IL-1β group were treated with 10 ng/mL IL-1β,while no treatment was given in Con group. The transcription levels of miR-16-5p and SOX5 mRNA in chondrocytes were detected by qRT-PCR;The contents of IL-6,TNF-αand IL-1β were detected by ELISA;The expression levels of Bcl-2,Bax and SOX5 protein were detected by Western blot and the apoptosis was detected by flow cytometry. Results Compared with IL-1 β group,the contents of IL-6,TNF-α and IL-1βin IL-1β + ALO-L group,IL-1β + ALO-M group and IL-1β + ALO-H group decreased significantly(t = 5. 002～20. 653,each P < 0. 001),the apoptosis rate decreased significantly(t = 5. 473～17. 371,each P < 0. 001),the expression level of Bcl-2 protein increased significantly(t = 7. 800～16. 100,each P < 0. 001),and the expression level of Bax protein decreased significantly(t = 4. 993～14. 311,each P < 0. 001);The mRNA transcription level of miR-16-5p gene increased significantly(t = 6. 688～16. 545,each P < 0. 001),while the mRNA transcription level and protein expression level of SOX5 gene decreased significantly(t = 4. 609～15. 393,each P < 0. 001). Compared with the IL-1β + miR-NC group,the mRNA transcription level of miR-16-5p in the IL-1β + miR-16-5p group increased significantly(t = 17. 106,P < 0. 001),the contents of IL-6,TNF-α and IL-1 β decreased significantly(t = 15. 030～20. 013,each P < 0. 001),the apoptosis rate decreased significantly(t = 12. 273,P < 0. 001),the expression level of Bcl-2 protein increased significantly(t = 15. 652,P < 0. 001),and the expression level of Bax protein decreased significantly(t = 12. 999,P < 0. 001). Compared with IL-1β +si-NC group,the expression level of SOX5(t = 13. 444,P < 0. 001),IL-6,TNF-α and IL-1β in IL-1β + si-SOX5 group decreased significantly(t = 14. 087～17. 103,each P < 0. 001),the apoptosis rate decreased significantly(t = 11. 991,P < 0. 001),the expression level of Bcl-2 protein increased significantly(t = 13. 864,P < 0. 001),and the expression level of Bax protein decreased significantly(t = 11. 818,P < 0. 001). Conclusion Alo inhibited the apoptosis of chondrocytes induced by IL-1β,thus reducing the injury of chondrocytes,of which the mechanism might be regulating the expression of miR-16-5p and SOX5 and the secretion of inflammatory factors in chondrocytes.

The effect of musk on cerebral ischemia-reperfusion in rats / Монголын Анагаах Ухаан

Introduction @#In recent years, there has been a significant increase of cerebrovascular disease in Mongolia, which is the second leading cause of mortality. There are dozens of Mongolian traditional medicine which is good efficiency for cerebral ischemia that contains musk.@*Aim@#Therefore, we aimed to investigate the effect of musk under the cerebral ischemia/reperfusion in rats.@*Materials and Methods@#Cerebral middle cerebral artery occlusion was established in male rat (90-minute occlusion followed by 24-hour reperfusion). Rats were divided into following groups: control group, ischemia group (cerebral ischemia and reperfusion), nimodipine administrated group (cerebral ischemia and reperfusion + treated with nimodipine), musk administrated group (cerebral ischemia and reperfusion + musk 50 mg/kg and 100 mg/kg). The brain tissue levels of IL-1β, TNF-α, IL-6, IL-10 cytokines were measured using enzyme linked immunosorbent assay (ELISA) every 1, 3, 7th days.@*Results@#Levels of cytokines (IL-1β, TNF-α and IL-6) were significantly lower in musk treated group compared to brain ischemia group (p<0.05). In contrast, treatment with musk was significantly improved neurological function with stimulation of M2 phenotype microglia cells and increased the anti-inflammatory cytokine level of IL-10 in the ischemic hemisphere of brain in rats@*Conclusion@#The mechanisms of musk are associated with increasing the brain tissue levels of IL-10, and reducing the levels of proinflammatory cytokines such as IL-1β, TNF-α, IL-6 subsequently stimulating neurogenesis and reduced ischemic zone. Musk may have neuroprotective effects against cerebral ischemia with stimulating M2 phenotype microglia cells in the brain. Regarding the ELISA, the effects of musk may be due to anti-inflammatory properties through inhibition of some of proinflammatory cytokines and stimulation of anti- inflammatory cytokines.

Effects of butterflybush flower eye drops on expression of inflammatory cytokines IL-1β, Mucin 5AC and P38MAPK in castrated male rabbits / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To observe the effects of butterflybush flower eye drops at different concentrations on expression of inflammatory crtokines IL-1β, Mucin 5AC(MUC5AC)and P38MAPK in castrated male rabbits, and to explore the therapeutic effect of that drops on dry eyes. <p>METHODS: Thirty-six male rabbits were randomly divided into blank group(A), model group(B), low concentrations butterflybush flower eye drops group(C, 1mg/mL), the medium concentrations drops group(D, 1.5mg/mL), the high concentrations drops group(E, 3mg/mL), and testosterone group(F). In addition to group A, the testes and epididymis were removed from each group to establish a dry eye animal model. After successful modeling, groups A and B remain unchanged. Groups C, D, and E were given different concentrations of butterflybush flower eye drops, 3 times/d. In group F, testosterone propionate was injected into the muscles of the thigh at a dose of 0.5mL/kg once every 3d. Fluorescein staining, Schirmer I test(SⅠt)and tear film break time(BUT)were measured under general anesthesia in each group, eatment. After 4wk of treatment, the rabbits were sacrificed and the conjunctival tissues of the eyes were taken. The expression of IL-1β, mucin 5AC and P38MAPK in the conjunctiva was detected by immunohistochemical staining.<p>RESULTS: Among low concentrations butterflybush flower eye drops group, the medium concentrations drops group and the high concentrations drops group, the SⅠt value was significantly higher than that of model group, and BUT was significantly longer than model group. The positive staining of corneal fluorescein was significantly improved compared with model group, which was statistically significant(<i>P</i><0.01). Among IL-1β and P38MAPK in the conjunctiva of high concentrations butterflybush flower eye drops group, the medium concentrations drops group and the low concentrations drops group, the positive expressions were lower than those in model group, and the expression of MUC5AC was higher than that in group model group(<i>P</i><0.01). In addition, the high concentrations drops group was superior to the low and the medium concentrations drops group.<p>CONCLUSION: Butterflybush flower eye drops have androgen-like effect. For castrated dry eyes of male rabbits, they can down-regulate the expression of IL-1beta and P38MAPK in dry conjunctival tissue and increase the expression of MUC5AC, thus reducing inflammation infiltration in dry conjunctival tissue and maintaining tear film stability, but their effect is weaker than that of androgen. To the treatment of dry eyes, the middle and high concentration groups of the drops had stronger effects than the low one, and the high concentration group was better than the medium one.

Increased platelet distribution width and reduced IL-2 and IL-12 are associated with thrombocytopenia in Plasmodium vivax malaria

BACKGROUND Thrombocytopenia in malaria involves platelet destruction and consumption; however, the cellular response underlying this phenomenon has still not been elucidated. OBJECTIVE To find associations between platelet indices and unbalanced Th1/Th2/Th17 cytokines as a response to thrombocytopenia in Plasmodium vivax infected (Pv-MAL) patients. METHODS Platelet counts and quantification of Th1/Th2/Th17 cytokine levels were compared in 77 patients with uncomplicated P. vivax malaria and 37 healthy donors from the same area (endemic control group - ENCG). FINDINGS Thrombocytopenia was the main manifestation in 55 patients, but was not associated with parasitaemia. The Pv-MAL patients showed increases in the mean platelet volume (MPV), which may be consistent with larger or megaplatelets. Contrary to the findings regarding the endemic control group, MPV and platelet distribution width (PDW) did not show an inverse correlation, due the increase in the heterogeneity of platelet width. In addition, the Pv-MAL patients presented increased IL-1β and reduced IL-12p70 and IL-2 serum concentrations. Furthermore, the reduction of these cytokines was associated with PDW values. MAIN CONCLUSIONS Our data demonstrate that an increase in MPV and the association between reductions of IL-2 and IL-12 and PDW values may be an immune response to thrombocytopenia in uncomplicated P. vivax malaria.

Study on the correlation between tear inflammatory factors and symptoms in patients with dry eye / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To observe the correlation between the interleukin-1β(IL-1β)and interleukin-18(IL-18)in tears of patients with dry eye and symptoms and signs.<p>METHODS: A total of 131 patients(262 eyes)who were treated for dry eye in the ophthalmology clinic of the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine were selected from September 2018 to December 12, and the patients were divided into dry eye classification standards according to the dry eye clinical consensus in 2013 No dry eye group, mild dry eye group, moderate dry eye group, severe dry eye group. All patients were examined for dry eye symptom score, break up time(BUT), and tear secretion test Schirmer I test(SⅠt), corneal fluorescein sodium staining(FL), ELISA method to detect the expression of IL-1β and IL-18 in tears, and to analyze the correlation between dry eye inflammatory factors and symptoms and signs.<p>RESULTS: There were significant differences in the expression of dry eye symptoms, BUT, SⅠt, FL and IL-1β and IL-18 in tears(<i>P</i><0.001), inflammatory factors IL-1β, IL-18 and dry eye symptom scores. FL was positively correlated(<i>P</i><0.05)and negatively correlated with BUT and SⅠt(<i>P</i><0.05).<p>CONCLUSION:Inflammatory factors in tears of dry eye patients were correlation with dry eye symptom and signsa.

Dose dependant anti-inflammatory effects of Dehydrocostus Lactone / Монголын Анагаах Ухаан

Introduction@#After central nervous system injury, microglia cells are activated to initiate inflammatory responses and release cytokines that beneficially or detrimentally affect surrounding cells. Lipopolysaccharide stimulates microglia cells and produce pro-inflammatory cytokines such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α. A dehydrocostus lactone (DDL) which is contained in medicinal plant, Saussurea lappa, is considered to have various health benefits in neurodegenerative diseases of central nervous system. </br> In this study, we aimed to investigate the anti-inflammatory effects of Dehydrocostus Lactone following lipopolysaccharide stimulation of microglial cells in vitro.@*Materials and Method@#The anti-inflammatory effects of dehydrocostus lactone were studied using lipopolysaccharide (LPS) stimulated murine microglia (BV2). BV2 were cultered in DMEM then three different doses (4µM, 8µM and 12µM) of DDL were added in the medium for 30 minutes respectively. Then BV2 were treated with 1 ng/ ml LPS for 24 hours to stimulate. The level of IL-1β, IL-6 and TNF-α were measured in 100µl of culturemedium supernatant by ELISA. Three different doses of DDL anti-inflammation groups (BV2+DDL+LPS), LPS-activated group (BV2+LPS) and control group (only BV2) were analysed. @*Results@#LPS-treated BV2 cells had increased IL-1β, IL-6 and TNF-α compared with those without LPS treatment. Pretreatment with dehydrocostus lactone prior to LPS treatment significantly decreased levels of IL-1β and TNF-α in a dose-dependent manner compared with LPS-treated BV2 cells and 4µM was the most effective anti-inflammatory dose of dehydrocostus lactone. As for IL-6, 12µM dehydrocostus lactone was the most effective anti-inflammatory dose, although all doses significantly decreased the level of IL-6, in a dose-dependent manner. @*Conclusion@#These results show that DDL decrease inflammation related IL-1β, IL-6 and TNF-α in a dose-dependent manner in microglia cells.

Effect of IL-1β on human synovial fluid-derived mesenchymal stem cells in the temporomandibular joint / 口腔疾病防治

Objective@#This study investigated the effects of interleukin-1β (IL-1β) on human synovial fluid-derived mesenchymal stem cells (hSFMSCs) in the temporomandibular joint.@*Methods @#hSFMSCs from synovial fluid samples of temporomandibular disorder (TMD) patients were cultured in vitro. hSFMSCs were divided into three groups with different concentrations of rhIL-1β in complete medium (α-MEM cell culture medium + 10% FBS + 1× GlutaMAX): 0 ng/mL IL-1β group, 1 ng/mL IL-1β group and 10 ng/mL IL-1β group. Changes in the rate of colony formation, growth curve, cell cycle and apoptosis of hSFMSCs under IL-1β stimulation were evaluated. The osteogenic, adipogenic and chondrogenic potential of the cells were also determined using histochemical and real-time fluorescence quantitative PCR methods. @*Results @#No significant differences in growth or proliferation capacity were observed in any IL-1β-stimulated group in comparisons of the colony-formation rate (F = 0.665, P=0.548), growth curve (F=0.001, P=0.999), cell cycle (FG1=0.773, PG1=0.503; FS =0.636, PS =0.562) or apoptosis (F=0.196, P=0.827) of the cells. However, the multidifferentiation capacity of hSFMSCs was affected in the inflammatory environment. Mineralized nodule and lipid clusters decreased significantly, and the gene expression levels of runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), peroxisomal proliferative receptor G2 (PPARG2) and lipoprotein lipase (LPL) were suppressed significantly in IL-1β-mediated induction medium (P < 0.05). In general, cartilage pellets formed in all the IL-1β-mediated chondrogenic differentiation groups. The gene levels of sex-determining region Y-related high-mobility group box-9 (SOX9) and collagen II were decreased (P < 0.05), while that of matrix metalloproteinase 13 (MMP13) was increased significantly in the presence of IL-1β (P < 0.05). @*Conclusion@# IL-1β directly affects the multidifferentiation potential of hSFMSCs but not their cell growth or proliferation ability.

Expression of Inflammasome Complex Following Various Oral Bacterial Infection in THP-1 Cells

Interleukin-1b (IL-1β), a proinflammatory cytokine, regulates the innate immune responses against bacterial infection. Mature IL-1β is produced from pro-IL-1β by activated caspase-1, which in turn is activated by the inflammasome complex formation. In this study, we compared the inflammasome mRNA expression induced by S. sanguinis, S. oralis, F. nucleatum and P. intermedia. Among the tested bacteria, S. sanguinis induced the highest IL-1β secretion. S. oralis, F. nucleatum and P. intermedia induced very weak IL-1β secretion. S. sanguinis mostly induced the NLRP3 mRNA expressions. Although F. nucleatum did not induce high IL-1β secretion, it induced high expression levels of AIM2, NLRP2, and NLRP3. No specific inflammasomes were induced by S. oralis and P.intermedia. Studying the inflammasome complex activation induced by oral bacteria may thus enhance our understanding of the pathogenesis of oral diseases.

Inhibitory activities of ginsenosides on the activation of NLRP3 inflammasome / 中国药科大学学报

@#To screen ginsenosides with inhibitory activities on the activation of NLRP3 inflammasome, 17 ginsenosides(PPT, PPD, Rb1, Rb2, Rb3, Rc, Rd, Re, Rg1, Rg2, Gg3, Rh1, Rh2, Ro, Rh2, Ro, cK, F1, F2)were used to treat mouse peritoneal macrophages(PMs)induced with NLRP3 inflammasome inducers(ATP, nigericin, or silica). In the present study, the higher anti-inflammatory activity of ginsenoside monomers were confirmed by analysis of IL-1β secretion in PMs. In the group of ATP induction, IL-1β secretion decreased after ginsenoside treatments, the high inhibitory effect was found at treatments of PPT, PPD, and F1(P< 0. 001). At the group of the nigericin, Rg3 also possessed higher inhibitory effect on IL-1β secretion except PPT, PPD, and F1(P< 0. 001). Furthermore, compared with groups of the ATP and nigericin, IL-1β secretion at the group of silica decreased after treatments of various ginsenoside monomers, including PPT, PPD, Rd, Rg3, Rb3, Rb2, F2, Re, F1, and Rh2(P< 0. 001). The present study indicates that PPT, PPD, and F1 can efficiently inhibit NLRP3 inflammasome activation induced by different inducers.

Biologic Response of Degenerative Living Human Nucleus Pulposus Cells to Treatment with Cytokines

PURPOSE: To investigate the molecular responses of various genes and proteins related to disc degeneration upon treatment with cytokines that affect disc-cell proliferation and phenotype in living human intervertebral discs (IVDs). Responsiveness to these cytokines according to the degree of disc degeneration was also evaluated. MATERIALS AND METHODS: The disc specimens were classified into two groups: group 1 (6 patients) showed mild degeneration of IVDs and group 2 (6 patients) exhibited severe degeneration of IVDs. Gene expression was analyzed after treatment with four cytokines: recombinant human bone morphogenic protein (rhBMP-2), transforming growth factor-beta (TGF-beta), interleukin-1beta (IL-1beta), and tumor necrosis factor-alpha (TNF-alpha). Molecular responses were assessed after exposure of cells from the IVD specimens to these cytokines via real-time polymerase chain reaction and immunofluorescence staining. RESULTS: mRNA gene expression was significantly greater for aggrecan, type I collagen, type II collagen, alkaline phosphatase, osteocalcin, and Sox9 in group 1 than mRNA gene expression in group 2, when the samples were not treated with cytokines. Analysis of mRNA levels for these molecules after morphogen treatment revealed significant increases in both groups, which were much higher in group 1 than in group 2. The average number of IVD cells that were immunofluorescence stained positive for alkaline phosphatase increased after treatment with rhBMP-2 and TGF-beta in group 1. CONCLUSION: The biologic responsiveness to treatment of rhBMP-2, TGF-beta, TNF-alpha, and IL-1beta in the degenerative living human IVD can be different according to the degree of degeneration of the IVD.

Interleukin-1beta Participates in the Development of Pneumococcal Acute Lung Injury and Death by Promoting Alveolar Microvascular Leakage

Streptococcus pneumoniae (S. pneumoniae, also known as pneumococcus) infections are major causes of death worldwide. Despite the development and use of effective antibiotics, high, early mortality due to pneumococcal infections has not been decreased for the last few decades. Recent study found a deadly hemorrhagic acute lung injury (ALI) as a major cause of death at the early stage of severe pneumococcal infections. Interleukin (IL)-1beta was known to play critical roles not only for the development of ALI but also resolution of it. The role of IL-1beta on the pathogenesis of pneumococcal ALI, however, has not been well understood yet. This study aims to investigate the role of IL-1beta on the development of pneumococcal ALI and subsequent death. IL-1beta expression was upregulated in the lungs of pneumococcal ALI in wild-type (WT) mice, but not in the plasma. Despite an increased expression of pulmonary IL-1beta, no inflammatory cell infiltration into airway has been observed. Upregulation of IL-1beta expression was indeed dependent on pneumococcal cytoplasmic toxin pneumolysin and its cell surface receptor Toll-like receptor 4. Deficiency of IL-1R1, a cell surface receptor of IL-1beta, resulted in a markedly reduced hemorrhagic pulmonary edema and early death in pneumococcal ALI. Finally, IL-1beta neutralization in WT mice protects against pulmonary hemorrhagic edema and death. These data suggest that pulmonary expression of IL-1beta exacerbates pneumolysin-induced ALI and death by promoting alveolar hemorrhagic edema.

Caspase-1 Independent Viral Clearance and Adaptive Immunity Against Mucosal Respiratory Syncytial Virus Infection

Respiratory syncytial virus (RSV) infection is recognized by the innate immune system through Toll like receptors (TLRs) and retinoic acid inducible gene I. These pathways lead to the activation of type I interferons and resistance to infection. In contrast to TLRs, very few studies have examined the role of NOD-like receptors in viral recognition and induction of adaptive immune responses to RSV. Caspase-1 plays an essential role in the immune response via the maturation of the proinflammatory cytokines IL-1beta and IL-18. However, the role of caspase-1 in RSV infection in vivo is unknown. We demonstrate that RSV infection induces IL-1beta secretion and that caspase-1 deficiency in bone marrow derived dendritic cells leads to defective IL-1beta production, while normal RSV viral clearance and T cell responses are observed in caspase-1 deficient mice following respiratory infection with RSV. The frequencies of IFN-gamma producing or RSV specific T cells in lungs from caspase-1 deficient mice are not impaired. In addition, we demonstrate that caspase-1 deficient neonatal or young mice also exhibit normal immune responses. Furthermore, we find that IL-1R deficient mice infected with RSV exhibit normal Th1 and cytotoxic T lymphocytes (CTL) immune responses. Collectively, these results demonstrate that in contrast to TLR pathways, caspase-1 might not play a central role in the induction of Th1 and CTL immune responses to RSV.

Los extractos acuoso y metanólico de Berberis darwinii H. (Berberidaceae) inhiben respuestas celulares innatas en monocitos humanos tratados in vitro / Aqueous and methanol extracts of Berberis darwinii H. (Berberidaceae) inhibit innate celular responses in human monocytes in vitro treated

Berberis darwinii H is a native plant of South America, popularly referred to Michay. This species has historically been used by indigenous cultures of Chile as medicinal herb. To preliminarily assess their anti-inflammatory effects was investigated the aqueous and methanolic root extract this plant in human monocytes. The results indicated that the extracts inhibit the production of superoxide anion, the expression of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1-beta (IL-1beta) in monocytes activated with lipopolysaccharide. This result suggests the existence of compounds with potential anti-inflammatory action in these extracts.

Berberis darwinii H. es una planta nativa de América del Sur, conocida popularmente como Michay. Esta especie ha sido históricamente utilizada por las culturas indígenas de Chile como hierba medicinal. Con el fin de evaluar preliminarmente sus efectos anti-inflamatorios, se investigaron dos tipos de extractos; metanólico y acuoso, preparados a partir de la raíz de esta planta. Los resultados indican que estos extractos inhiben la producción de anión superóxido, la expresión del factor de necrosis tumoral-alfa (TNF-alfa) y de interleucina-1beta, (IL-1beta) en monocitos activados con lipopolisacárido. Estos resultados sugieren la existencia de compuestos con potencial acción antiinflamatoria en esta planta.

Nuclear Factor-kappaB Dependent Induction of TNF-alpha and IL-1beta by the Aggregatibacter actinomycetemcomitans Lipopolysaccharide in RAW 264.7 Cells

Aggregatibacter actinomycetemcomitans is an important pathogen in the development of localized aggressive periodontitis. Lipopolysaccharide (LPS) is a virulent factor of periodontal pathogens that contributes to alveolar bone loss and connective tissue degradation in periodontal disease. Our present study was designed to investigate the cytokine expression and signaling pathways regulated by A. actinomycetemcomitans LPS (Aa LPS). Cytokine gene expression profiling in RAW 264.7 cells was performed by microarray analyses. The cytokine mRNA and protein levels and related signaling pathways induced by Aa LPS were measured by RT-PCR, ELISA and western blotting. Microarray results showed that Aa LPS strongly induced the expression of NF-kappaB, NF-kappaB-related genes, inflammatory cytokines, TNF-alpha and IL-1beta in RAW 264.7 cells. NF-kappaB inhibitor pretreatment significantly reduced the levels of TNF-alpha and IL-1beta mRNA and protein. In addition, the Aa LPS-induced TNF-alpha and IL-1beta expression was inhibited by p38/JNK MAP kinase inhibitor pretreatment. These results show that Aa LPS stimulates TNF-alpha and IL-1beta expression through NF-kappaB and p38/JNK activation in RAW 264.7 cells, suggesting the essential role of this pathway in the pathogenesis of localized aggressive periodontitis.

Effect of bamboo salt on human gingival fibroblasts / 대한구강보건학회지

OBJECTIVES: To evaluate the anti-inflammatory and cytotoxic activities of bamboo salt. METHODS: Cytotoxicity of bamboo salt and bay salt (0.01%, 0.1%, and 1%) was evaluated using MTT assay. In addition, secretion of the pro-inflammatory cytokines interleukin (IL)-1beta and IL-6 from human gingival fibroblasts (HGFs) was measured after application of 0.01% and 0.1% concentrations by using real-time polymerase chain reaction. RESULTS: Bamboo salt and bay salt at 1% concentration were cytotoxic to HGFs at 24 h; however, no such effect was observed at 0.01% or 0.1%. Bamboo salt showed a relatively low inhibitory effect. IL-1beta secretion was inhibited by a 0.1% solution of bamboo salt. IL-6 secretion was inhibited by both bamboo salt and bay salt at 0.1% concentration. CONCLUSIONS: The above results suggest that bamboo salt inhibits the release of IL-1beta and IL-6 from HGFs. Thus, bamboo salt may be a useful material for gingival inflammation.

alpha-Asarone Ameliorates Memory Deficit in Lipopolysaccharide-Treated Mice via Suppression of Pro-Inflammatory Cytokines and Microglial Activation

alpha-Asarone exhibits a number of pharmacological actions including neuroprotective, anti-oxidative, anticonvulsive, and cognitive enhancing action. The present study investigated the effects of alpha-asarone on pro-inflammatory cytokines mRNA, microglial activation, and neuronal damage in the hippocampus and on learning and memory deficits in systemic lipopolysaccharide (LPS)-treated C57BL/6 mice. Varying doses of alpha-asarone was orally administered (7.5, 15, or 30 mg/kg) once a day for 3 days before the LPS (3 mg/kg) injection. alpha-Asarone significantly reduced TNF-alpha and IL-1beta mRNA at 4 and 24 hours after the LPS injection at dose of 30 mg/kg. At 24 hours after the LPS injection, the loss of CA1 neurons, the increase of TUNEL-labeled cells, and the up-regulation of BACE1 expression in the hippocampus were attenuated by 30 mg/kg of alpha-asarone treatment. alpha-Asarone significantly reduced Iba1 protein expression in the hippocampal tissue at a dose of 30 mg/kg. alpha-Asarone did not reduce the number of Iba1-expressing microglia on immunohistochemistry but the average cell size and percentage areas of Iba1-expressing microglia in the hippocampus were significantly decreased by 30 mg/kg of alpha-asarone treatment. In the Morris water maze test, alpha-asarone significantly prolonged the swimming time spent in the target and peri-target zones. alpha-Asarone also significantly increased the number of target heading and memory score in the Morris water maze. The results suggest that inhibition of pro-inflammatory cytokines and microglial activation in the hippocampus by alpha-asarone may be one of the mechanisms for the alpha-asarone-mediated ameliorating effect on memory deficits.

alpha-Asarone Ameliorates Memory Deficit in Lipopolysaccharide-Treated Mice via Suppression of Pro-Inflammatory Cytokines and Microglial Activation

alpha-Asarone exhibits a number of pharmacological actions including neuroprotective, anti-oxidative, anticonvulsive, and cognitive enhancing action. The present study investigated the effects of alpha-asarone on pro-inflammatory cytokines mRNA, microglial activation, and neuronal damage in the hippocampus and on learning and memory deficits in systemic lipopolysaccharide (LPS)-treated C57BL/6 mice. Varying doses of alpha-asarone was orally administered (7.5, 15, or 30 mg/kg) once a day for 3 days before the LPS (3 mg/kg) injection. alpha-Asarone significantly reduced TNF-alpha and IL-1beta mRNA at 4 and 24 hours after the LPS injection at dose of 30 mg/kg. At 24 hours after the LPS injection, the loss of CA1 neurons, the increase of TUNEL-labeled cells, and the up-regulation of BACE1 expression in the hippocampus were attenuated by 30 mg/kg of alpha-asarone treatment. alpha-Asarone significantly reduced Iba1 protein expression in the hippocampal tissue at a dose of 30 mg/kg. alpha-Asarone did not reduce the number of Iba1-expressing microglia on immunohistochemistry but the average cell size and percentage areas of Iba1-expressing microglia in the hippocampus were significantly decreased by 30 mg/kg of alpha-asarone treatment. In the Morris water maze test, alpha-asarone significantly prolonged the swimming time spent in the target and peri-target zones. alpha-Asarone also significantly increased the number of target heading and memory score in the Morris water maze. The results suggest that inhibition of pro-inflammatory cytokines and microglial activation in the hippocampus by alpha-asarone may be one of the mechanisms for the alpha-asarone-mediated ameliorating effect on memory deficits.

Transglutaminase-2 Is Involved in Expression of Osteoprotegerin in MG-63 Osteosarcoma Cells

Osteoprotegerin (OPG) is a secreted glycoprotein and a member of the tumor necrosis factor receptor superfamily. It usually functions in bone remodeling, by inhibiting osteoclastogenesis through interaction with a receptor activator of the nuclear factor kappaB (RANKL). Transglutaminases-2 (Tgase-2) is a group of multifunctional enzymes that plays a role in cancer cell metastasis and bone formation. However, relationship between OPG and Tgase-2 is not studied. Therefore, we investigated the involvement of 12-O-Tetradecanoylphorbol 13-acetate in the expression of OPG in MG-63 osteosarcoma cells. Interleukin-1beta time-dependently induced OPG and Tgase-2 expression in cell lysates and media of the MG-63 cells by a Western blot. Additional 110 kda band was found in the media of MG-63 cells. 12-O-Tetradecanoylphorbol 13-acetate also induced OPG and Tgase-2 expression. However, an 110 kda band was not found in TPA-treated media of MG-63 cells. Cystamine, a Tgase-2 inhibitor, dose-dependently suppressed the expression of OPG in MG-63 cells. Gene silencing of Tgase-2 also significantly suppressed the expression of OPG in MG-63 cells. Next, we examined whether a band of 110 kda of OPG contains an isopeptide bond, an indication of Tgase-2 action, by monoclonal antibody specific for the isopeptide bond. However, we could not find the isopeptide bond at 110 kda but 77 kda, which is believed to be the band position of Tgase-2. This suggested that 110 kda is not the direct product of Tgase-2's action. All together, OPG and Tgase-2 is induced by IL-1beta or TPA in MG-63 cells and Tgase-2 is involved in OPG expression in MG-63 cells.

The Effect of Agmatine on Expression of IL-1beta and TLX Which Promotes Neuronal Differentiation in Lipopolysaccharide-Treated Neural Progenitors

Differentiation of neural progenitor cells (NPCs) is important for protecting neural cells and brain tissue during inflammation. Interleukin-1 beta (IL-1beta) is the most common pro- inflammatory cytokine in brain inflammation, and increased IL-1beta levels can decrease the proliferation of NPCs. We aimed to investigate whether agmatine (Agm), a primary polyamine that protects neural cells, could trigger differentiation of NPCs by activating IL-1beta in vitro. The cortex of ICR mouse embryos (E14) was dissociated to culture NPCs. NPCs were stimulated by lipopolysaccharide (LPS). After 6 days, protein expression of stem cell markers and differentiation signal factors was confirmed by using western blot analysis. Also, immunocytochemistry was used to confirm the cell fate. Agm treatment activated NPC differentiation significantly more than in the control group, which was evident by the increased expression of a neuronal marker, MAP2, in the LPS-induced, Agm-treated group. Differentiation of LPS-induced, Agm-treated NPCs was regulated by the MAPK pathway and is thought to be related to IL-1beta activation and decreased expression of TLX, a transcription factor that regulates NPC differentiation. Our results reveal that Agm can promote NPC differentiation to neural stem cells by modulating IL-1beta expression under inflammatory condition, and they suggest that Agm may be a novel therapeutic strategy for neuroinflammatory diseases.