Serum HMGB1 Levels Research Articles

Fingolimod anti-inflammatory and neuroprotective effects modulation of RAGE axis in multiple sclerosis patients

BackgroundWe investigated Fingolimod treatment effects on the RAGE (receptor for advanced glycation endproducts) axis in multiple sclerosis (MS) patients. The primary outcome of the study was whether Fingolimod treatment increases serum levels of the soluble RAGE isoforms, sRAGE and esRAGE – both being considered putative endogenous inhibitors of RAGE signaling. Additional variables were serum levels of RAGE ligands, the high mobility group box (HMGB)1 and pentosidine. MethodsSerum levels of the study variables were measured by ELISA, and compared between baseline (before Fingolimod treatment) and 6 and 12 months post-drug treatment in 17 relapsing MS patients. Fingolimod treatment effects on MS disease progression were assessed by comparing pre- and post-Fingolimod values of the EDSS and rate of clinical relapse, and changes in the T1-and T2-enahncing lesions on the MRI scan.methods ResultsTwelve months treatment with Fingolimod increased serum levels of sRAGE and esRAGE by 32.4% (P = 0.004) and 48.5% (P = 0.007) respectively. In addition, Fingolimod treatment reduced serum levels of HMGB1 by 71.6% (P = 0.02) and pentosidine serum levels by 41.3% (P = 0.12). EDSS remained stable (baseline: 3.57 ± 1.56; post-Fingolimod: 3.54 ± 1.2, P = 0.96) and the rate of clinical relapse decreased near significantly (P = 0.094). T1-and T2-enhancing lesions remained stable, showing no significant changes pre-vs. post-Fingolimod treatment. ConclusionFingolimod mediates modulation of the RAGE axis which apparently contributes to the Fingolimod's anti-inflammatory and neuroprotective effects. These findings may provide a rationale for the clinical efficacy of Fingolimod in pathological states other than MS, where dysregulation of the RAGE axis plays a role.

Oxidized low-density lipoprotein promotes osteoclast differentiation from CD68 positive mononuclear cells by regulating HMGB1 release

Patients with ankylosing spondylitis (AS) have an increased risk for cardiovascular mortality. The circulating ox-LDL/LDL ratio is associated with subclinical atherosclerosis in patients with systemic lupus erythematosus. In this study, we found that the ox-LDL/LDL ratio was increased in AS patients. The levels of serum RANKL and HMGB1 were also elevated in AS patients, and the number of CD68+/RANK+ cells was increased in peripheral blood from AS patients. 0.03% ox-LDL in LDL, similar to the ox-LDL/LDL ratio in peripheral blood from AS patients, promoted cytoplasmic translocation and release of HMGB1 as well as RANK expression. Further investigation evidenced that ox-LDL-induced EGR1 expression contributed to the cytoplasmic translocation of HMGB1 and CD68 assisted the secretion of HMGB1 from cytoplasm to extracellular matrix. Extracellular HMGB1 induced RANK expression in CD68+ mononuclear cells, forming osteoclast precursors that were differentiated to osteoclasts in response to RANKL. Taken together, these results suggested that the changes, including ox-LDL/LDL ratio, CD68+/RANK+ cells number, and the levels of RANKL and HMGB1 in AS patients, favored osteoclastogenesis.

Serum high mobility group box 1 protein levels are not associated with either histological severity or treatment response in children and adults with nonalcoholic fatty liver disease.

AimSerum high mobility group box 1 protein (HMGB1) is a proinflammatory molecule that could potentially serve as a biomarker for non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) due to its correlation with degree of liver fibrosis. The aim of the current study was to examine the cross-sectional and longitudinal relationships between serum HMGB1 levels and liver histology in adults and children with NAFLD participating in two large randomized controlled trials.MethodsSerum HMGB1 levels were measured at various time points in adults and children with NAFLD, who participated in PIVENS and TONIC clinical trials respectively. PIVENS trial compared vitamin E or pioglitazone to placebo in adults whereas TONIC trial compared vitamin E or metformin to placebo in children. Participants had liver biopsies at baseline and the end of treatment (96 weeks), and liver histology was reviewed by a central committee of study pathologists.ResultsIn the cross-sectional analyses (n = 205 for PIVENS and 109 for TONIC), there was no significant relationship between serum HMGB1 levels and histological features such as steatosis, ballooning, inflammation, fibrosis, or presence of steatohepatitis in either adults or children. Serum HMGB1 levels did not change significantly during treatment either with placebo, vitamin E therapy (P = 0.81) or pioglitazone (P = 0.09) in the PIVENS trial. Similarly, serum HMGB1 levels did not change significantly during treatment either with placebo, metformin (P = 0.15) or vitamin E (P = 0.23) in the TONIC trial. In the longitudinal analyses (n = 105 for PIVENS and 109 for TONIC), changes in serum HMGB1 levels did not correlate with histologic improvement or resolution of NASH in either adults or children. There was no relationship between serum HMGB1 and ALT levels in either adults or children with NAFLD.ConclusionSerum HMGB1 levels were not associated with histological severity or treatment response in either children or adults with NAFLD.

Serum high mobility group box-1 and osteoprotegerin levels are associated with peripheral arterial disease and critical limb ischemia in type 2 diabetic subjects

BackgroundHigh mobility group box-1 (HMGB-1) is a nuclear protein also acting as inflammatory mediator, whilst osteoprotegerin (OPG), member of tumor necrosis factor receptor superfamily, is indicated as marker of vascular calcification. Peripheral artery disease (PAD) and type 2 diabetes (T2D) are clinical conditions characterized by elevated serum inflammatory markers and vascular calcification enhancement. The aim of this study was to investigate the potential role of HMGB-1, OPG and several inflammatory mediators such as C-reactive protein (HsCRP), tumor necrosis factor-alpha and interleukin-6 (IL-6) on the presence and severity of peripheral artery disease in patients with T2D.MethodsIn this retrospective observational study, we have analyzed HMGB-1, OPG and inflammatory cytokines serum levels in 1393 type 2 diabetic patients with PAD and without PAD (WPAD).ResultsHMGB-1 (7.89 ± 15.23 ng/mL), OPG (6.54 ± 7.76 pmol/L), HsCRP (15.6 ± 14.4 mg/L) and IL-6 (56.1 ± 28.6 pg/mL) serum levels were significantly higher in patients with PAD than in those WPAD (3.02 ± 8.12 ng/mL, P ˂ 0.001; 2.98 ± 2.01 pmol/L, P < 0.001; 7.05 ± 4.4 mg/L, P < 0.001; 37.5 ± 20.2 pg/mL, P < 0.001 respectively). Moreover HMGB-1 (P < 0.001), OPG (P < 0.001), HsCRP (P < 0.001) and IL-6 (P < 0.001) serum levels were positively correlated with clinical severity of PAD. HMGB-1 (adjusted OR 12.32; 95% CI 3.56–23.54, P = 0.023) and OPG (adjusted OR 3.53; 95% CI 1.54–6.15, P = 0.019) resulted independent determinants of PAD in patients with T2D after adjusting for the conventional cardiovascular risk factor and established inflammatory mediators.ConclusionsIn T2D patients HMGB-1 and OPG serum levels are higher in patients affected by PAD and independently associated with its occurrence and clinical severity.

Intestinal Permeability Biomarker Zonulin is Elevated in Healthy Aging

IntroductionIncreased gut permeability (“leaky gut”) has been proposed as a potential contributor to age-related inflammation and gut dysbiosis. However, information on the relationship between a leaky gut and inflammation and physical frailty during aging are limited. ObjectiveTo investigate the hypothesis that an aging-associated leaky gut is linked to the age-related inflammation and frailty. MethodsTwo cohorts of healthy adults were studied: young (18–30 years old, n = 19) and older (≥70 years old, n = 18). Serum concentrations of the tumor necrosis factor (TNF)-α and interleukin (IL)-6, zonulin (a marker for leaky gut), and high-mobility group box protein (HMGB1, a nuclear protein triggering inflammation) were measured. Correlations of serum levels of zonulin and HMGB1 with strength of plantar flexor muscles and number of steps taken per day were analyzed. ResultsSerum concentration of zonulin and HMGB1 were 22% (P = .005) and 16% (P = .010) higher in the older versus young adults. Serum zonulin was positively associated with concentrations of TNF-α (r = 0.357, P = .032) and IL-6 (r = 0.345, P = .043). Importantly, both zonulin and HMGB1 were negatively correlated with skeletal muscle strength (zonulin: r = −0.332, P = .048; HMGB1: r = −0.383, P = .023), and habitual physical activity (zonulin: r = −0.410, P = .016; HMGB1: r = −0.483, P = .004). ConclusionsSerum zonulin was associated with both systemic inflammation and 2 key indices of physical frailty. These data suggest that a leaky gut may play a critical role in the development of age-related inflammation and frailty.

Study on applications of rheum sterile solution in rats following sepsis and effect on inflammatory factors

Objective To study the therapeutic effect of rheum(Chinese herbal medicine) preparation made by using ultrasonic technique on pro-inflammatory cytokines and sepsis in rats. In order to offer novel measure for the treatment of critically ill patients. Methods Firstly, rheum sterile solution was prepared through ultrasonic technique. Secondly, fifty healthy male SD rats were randomly(random number) divided to CLP group and rheum group. Moderate degree of sepsis model was established by using cecal ligation and puncture(CLP). Rats in group rheum received the liquid rheumpreparation via intragastric administration, while rats in group CLP received saline instead. The 7-day survival rate was recorded and was compared between two groups. In addition, another fifty-four rats were randomly(random number) divided to sham group, CLP group and rheum group(n=18 in each group). CLP was performed to induce sepsis in CLP group and rheum group. Then rats in rheum group received rheum sterile solution via intragastric administration, while rats in CLP group received saline instead. At 12 hours, 24 hours and 48 hours after modeling, six rats in each group were randomly sacrificed. Serum TNF-α and HMGB1 levels were detected by ELISA method. Levels of RAGE, HMGB1 and NF-κB P65 in small intestine were detected by Western Blot. Results Level of anthraquinones extracted from rheum by ultrasonic technique was higher than that by conwentional decoction method. The 7-day survival rate of rats in rheum group(76%) was higher than that in CLP group(48%)(P 0.05). At 24 hours and 48 hours after modeling, serum HMGB1 levels were significantly lower in rheum group than those in CLP group(P<0.05). Compared with sham group, protein levels of HMGB1, RAGE and NF-κB in small intestine were elevated in CLP group and rheum group at 48 hours after modeling(P<0.01), while protein levels of above biomarker were higher in CLP group than those in rheum group(P<0.05). Conclusions Rheum sterile solution could down-regulate the level of pro-inflammatory cytokines, modulate the inflammatory response, and improve the survival rate in rats with sepsis. Key words: Ultrasonic technique; Rheum sterile solution; Stability; Sepsis; Proinflammatory cytokines

Esmolol reduces apoptosis and inflammation in early sepsis rats with abdominal infection

BackgroundEsmolol is a highly selective beta 1 receptor blocker with various effects such as slowing heart rate, lowering blood pressure and reducing myocardial oxygen consumption. However, few studies have reported the use of beta blockers in sepsis with multiple organ dysfunctions. This study aimed to investigate the effects of esmolol on reducing apoptosis and inflammation in early sepsis rats with abdominal infection. MethodsRats were randomly divided into sham operation group, sepsis group, antibiotic group, Esmolol + antibiotic group with low, median and high dose Esmolol (L group, M group and H group). Values between two or more groups were compared by independent t-tests. ResultsIn the liver and kidney, we found inflammatory infiltration in sepsis group while pathological aspects reduced in L, M and H groups. Bcl-2 mRNA and protein levels increased while Bax mRNA and protein levels decreased in the liver and kidney of L, M and H groups. Serum IL-6, HMGB-1 and TNF-α levels decreased but IL-10 level increased in L, M and H groups, compared to sepsis group. Compared to sepsis and antibiotic groups, the levels of myocardial enzymes were lower in L, M and H groups. ConclusionThe administration of esmolol in early sepsis may reduce inflammation, inhibit apoptosis and protect key organs.

MP06-13 BENEFICAL EFFECT OF HYDROGEN SULFIDE ON RENAL ISCHEMIA-REPERFUSION INJURY IN CLAWN MINIATURE SWINE

You have accessJournal of UrologyTransplantation & Vascular Surgery: Renal Transplantation & Vascular Surgery I1 Apr 2017MP06-13 BENEFICAL EFFECT OF HYDROGEN SULFIDE ON RENAL ISCHEMIA-REPERFUSION INJURY IN CLAWN MINIATURE SWINE Yuichi Ariyoshi, Mitsuhiro Sekijima, Takahiro Murokawa, Hisashi Sahara, Motoo Araki, Yasutomo Nasu, and Kazuhiko Yamada Yuichi AriyoshiYuichi Ariyoshi More articles by this author , Mitsuhiro SekijimaMitsuhiro Sekijima More articles by this author , Takahiro MurokawaTakahiro Murokawa More articles by this author , Hisashi SaharaHisashi Sahara More articles by this author , Motoo ArakiMotoo Araki More articles by this author , Yasutomo NasuYasutomo Nasu More articles by this author , and Kazuhiko YamadaKazuhiko Yamada More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.258AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The organ shortage is the major problem in kidney transplantation. To overcome this obstacle, kidney transplants of the marginal donor, cardiac death (DCD) and extended-criteria donors, were increasing. In this setting, the prevention of ischemia-reperfusion injury (IRI) is important for both early and long-term allograft function. Hydrogen Sulfides (H2S) have recently been reported to demonstrate both anti-inflammatory and cytoprotective effects. However, the efficacy and safety of H2S has yet to be elicited in a large animal model. We investigated whether H2S administration was effective for control of renal IRI and optimal administration method in a large animal model. METHODS Female, MHC-inbred, CLAWN miniature swine (n=8) underwent renal ischemia for 120-minutes by occlusion of the left renal artery and vein. Group 1 animals (n=3) underwent renal ischemia exclusively without any additional treatment. Group 2 recipients (n=2) received 1.1 mg/kg of intravenous Na2S 10-minutes prior to kidney reperfusion, followed by an additional 1.1 mg/kg of Na2S 30-minutes post-reperfusion. Group 3 recipients (n=3) underwent selective renal artery administration of 1.1 mg/kg of Na2S 10-minutes prior to reperfusion, followed by an additional 1.1 mg/kg of Na2S 30-minutes post-reperfusion via the supra-renal aorta with concomitant occlusion of the infra-renal aorta, thus allowing for exclusive renal administration. Post-operative renal function was monitored by daily serum creatinine, analysis of circulating cytokine activity (TNF-α, IL-6 and HMGB1) to measure the inflammatory response to IRI and histological evaluation of renal biopsies obtained on post-operative days (POD) 2, 7 and 14. RESULTS H2S administration did not result in any adverse side effects in the recipients. All animals experienced transient acute kidney injury, achieving a peak serum creatinine level by POD 3. Recipients in the untreated group had the higher post-operative serum creatinine level than selective renal H2S administration group (POD4: 4.4 vs 8.6 mg/dl, p<0.001). Pathologically, Control group still showed strongly epithelial flattering and vacuolations, congestion of PTC, and condensed tubular nuclei as a result of influence of IRI. In systemic group, the structures of renal tubule were well preserved comparatively, and those changes in selective group were obviously disappeared. Furthermore, on POD 7, specimens from selective group showed disappointment of IRI change. In selective group, the serum TNF-α and HMGB-1 level was lower than systemic group (TNF-α: 30.6 vs 85.4 pg/ml, p<0.05, HMGB-1: 2.6 vs 11.4 ng/ml, p<0.05). The mRNA expression of IL-1β was also suppressed in selective group specimen. CONCLUSIONS H2S administration performed safety and appeared to have potential cytoprotective and anti-inflammatory effects following renal IRI. This effect was most profound with selective renal artery administration. Further work investigating the benefits of H2S for organ procurement and preservation is warranted as this may allow for improved outcomes following renal transplantation. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e77-e78 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Yuichi Ariyoshi More articles by this author Mitsuhiro Sekijima More articles by this author Takahiro Murokawa More articles by this author Hisashi Sahara More articles by this author Motoo Araki More articles by this author Yasutomo Nasu More articles by this author Kazuhiko Yamada More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

HMGB1 targeting by ethyl pyruvate suppresses malignant phenotype of human mesothelioma.

Human malignant mesothelioma (MM) is an aggressive cancer linked to asbestos and erionite exposure. We previously reported that High-Mobility Group Box-1 protein (HMGB1), a prototypic damage-associated molecular pattern, drives MM development and sustains MM progression. Moreover, we demonstrated that targeting HMGB1 inhibited MM cell growth and motility in vitro, reduced tumor growth in vivo, and prolonged survival of MM-bearing mice. Ethyl pyruvate (EP), the ethyl ester of pyruvic acid, has been shown to be an effective HMGB1 inhibitor in inflammation-related diseases and several cancers. Here, we studied the effect of EP on the malignant phenotype of MM cells in tissue culture and on tumor growth in vivo using an orthotopic MM xenograft model. We found that EP impairs HMGB1 secretion by MM cells leading to reduced RAGE expression and NF-κB activation. As a consequence, EP impaired cell motility, cell proliferation, and anchorage-independent growth of MM cells. Moreover, EP reduced HMGB1 serum levels in mice and inhibited the growth of MM xenografts.Our results indicate that EP effectively hampers the malignant phenotype of MM, offering a novel potential therapeutic approach to patients afflicted with this dismal disease.

Altered balance of epidermis-related chemokines in epidermolysis bullosa

BackgroundEpidermolysis bullosa (EB) is a congenital, refractory skin disease and there are no fundamental treatments. Recently, allogenic cell therapies are beginning to be applied as potential treatments, that are based on the concept that the allogenic cells can migrate into the skin and reconstitute the skin components. Although the mechanisms of cell migration into skin are not fully understood, chemokines are regarded as key factors in recruiting bone marrow-derived cells. ObjectivesOur study aims to elucidate the expression of chemokines in the EB patients. MethodsWe determined the expression of wound-healing related chemokines in the sera, keratinocytes, and skin tissues of EB patients and compared them to those of healthy volunteers by enzyme-linked immunosorbent assays, quantitative reverse transcription PCR, and immunofluorescence staining. ResultsThe serum levels of CXCL12 and HMGB1 were found to be significantly elevated in the EB patients. Conversely, the serum levels of CCL21 were found to be lower in the EB patients than in healthy controls. In addition, the serum levels of CXCL12 tended to increase and the serum levels of CCL27 tended to decrease with an increase in the affected body surface areas. To detect the origin of the circulating chemokines, we performed immunofluorescence staining. CCL21, CCL27, HMGB1 and CXCL12 were stained more broadly in the EB patient tissues than those in the control tissues. ConclusionsThese results suggest that fluctuations in chemokine levels may contribute in a coordinated way to the wound-healing process and lend clues toward efficient cell therapies for EB.

Elevated Serum Level of HMGB1 in Patients with the Antiphospholipid Syndrome.

Pregnancy problems are common in patients with rheumatic disease; indeed, autoimmune disorders and autoantibodies can affect pregnancy progress and lead to maternal complications. Recent studies have highlighted a close association between HMGB1, chronic inflammation, and autoimmune diseases. Thus, in this investigation, we analyzed serum levels of HMGB1, an alarmin which plays a pivotal role in inducing and enhancing immune cell function. Sera from 30 patients with antiphospholipid syndrome (11 primary and 19 secondary APS), 35 subjects with pregnancy morbidity, and 30 healthy women were analysed for HMGB1 and its putative receptor RAGE (sRAGE) by Western blot and for TNF-α by ELISA. Results revealed that APS patients showed significantly increased serum levels of HMGB1, sRAGE, and the proinflammatory cytokine TNF-α, as compared to healthy women. However, also, the pregnancy morbidity subjects showed significantly increased levels of HMGB1 and sRAGE as well as TNF-α compared to healthy women. Our findings suggest that in subjects with pregnancy morbidity, including obstetric APS, elevated levels of HMGB1/sRAGE may represent an alarm signal, indicating an increase of proinflammatory triggers. Further studies are needed to evaluate the role of HMGB1/sRAGE as a possible tool to evaluate the risk stratification of adverse pregnancy outcomes.

Serum Level of HMGB1 Protein and Inflammatory Markers in Patients with Secondary Peritonitis: Time Course and the Association with Clinical Status.

SummaryBackgroundIntra-abdominal infection in secondary peritonitis drives as excessive production of inflammatory mediators and the development of systemic inflammatory response syndrome (SIRS) or sepsis. Finding a specific marker to distinguish SIRS from sepsis would be of immense clinical importance for the therapeutic approach. It is assumed that high-mobility group box 1 protein (HMGB1) could be such a marker. In this study, we examined the time course changes in the blood levels of HMGB1, C-reactive protein (CRP), procalcitonin (PCT) and serum amyloid A (SAA) in patients with secondary peritonitis who developed SIRS or sepsis.MethodsIn our study, we evaluated 100 patients with diffuse secondary peritonitis who developed SIRS or sepsis (SIRS and SEPSIS group) and 30 patients with inguinal hernia as a control group. Serum levels of HMGB1, CRP, PCT, and SAA were determined on admission in all the patients, and monitored daily in patients with peritonitis until discharge from hospital.ResultsPreoperative HMGB1, CRP, PCT and SAA levels were statistically highly significantly increased in patients with peritonitis compared to patients with inguinal hernia, and significantly higher in patients with sepsis compared to those with SIRS. All four inflammatory markers changed significantly during the follow-up. It is interesting that the patterns of change of HMGB1 and SAA over time were distinctive for SIRS and SEPSIS groups.ConclusionsHMGB1 and SAA temporal patterns might be useful in distinguishing sepsis from noninfectious SIRS in secondary peritonitis.

Down-Regulation of Serum High-Mobility Group Box 1 Protein in Patients with Pulmonary Tuberculosis and Nontuberculous Mycobacterial Lung Disease.

BackgroundRecently, increased levels of high-mobility group box 1 protein (HMGB1) have been identified in various inflammatory conditions and infections. However, no studies have evaluated the HMGB1 level in nontuberculous mycobacterial (NTM) lung disease, and compared it to mycobacterial lung disease.MethodsA total of 60 patients newly diagnosed with NTM lung disease, 44 culture-positive pulmonary tuberculosis (TB) patients, and 34 healthy controls, were included in this study. The serum HMGB1 concentrations were quantified using HMGB1 enzyme-linked immunosorbent assay kits.ResultsSerum HMGB1 level in patients with pulmonary TB or NTM lung disease, was significantly lower than that of the healthy controls. In addition, the serum HMGB1 level in TB patients was significantly lower than patients with NTM lung disease. However, the levels in NTM patient subgroups did not differ according to the causative species, disease progression, and disease phenotype.ConclusionAlthough low levels of serum HMGB1 has the potential to be a marker of mycobacterial lung disease, these levels were unable to differentiate disease progression and disease phenotype in NTM lung diseases.

Glycyrrhizin Protects Rats from Sepsis by Blocking HMGB1 Signaling.

Background. HMGB1 acts as an important inflammatory mediator and is a potential therapeutic target for sepsis. Glycyrrhizin (GL), a natural triterpene glycoside derived from licorice, has been demonstrated to inhibit HMGB1 activity. The aim of this study is to explore how GL affects the HMGB1 signaling in sepsis. Methods. We used a CLP model of sepsis and in vitro LPS or HMGB1-treated NR8383 cells to examine the effects of GL on expression of HMGB1 and proinflammatory cytokines. Furthermore, we explored the effect of GL on interactions between HMGB1 and RAGE or TLR4 and the activations of NF-κB and MAPKs. Results. GL significantly decreased mortality and reduced serum levels of HMGB1 in vivo. GL also attenuated the release and expression of HMGB1 and proinflammatory cytokines. Direct stimulation by HMGB1 elevated the release of proinflammatory cytokines faster than LPS did and it was also inhibited by GL. Furthermore, GL blocked the interaction of HMGB1 with RAGE and TLR4 and suppressed the downstream MAPKs/NF-κB signaling pathway. Conclusion. GL may protect rats against sepsis by blocking the interaction of HMGB1 with cell surface receptors and HMGB1-mediated inflammatory responses.

Salidroside Inhibits HMGB1 Acetylation and Release through Upregulation of SirT1 during Inflammation.

HMGB1, a highly conserved nonhistone DNA-binding protein, plays an important role in inflammatory diseases. Once released to the extracellular space, HMGB1 acts as a proinflammatory cytokine that triggers inflammatory reaction. Our previous study showed that salidroside exerts anti-inflammatory effect via inhibiting the JAK2-STAT3 signalling pathway. However, whether salidroside inhibits the release of HMGB1 is still unclear. In this study, we aim to study the effects of salidroside on HMGB1 release and then investigate the potential molecular mechanisms. In an experimental rat model of sepsis caused by CLP, salidroside administration significantly attenuated lung injury and reduced the serum HMGB1 level. In RAW264.7 cells, we investigated the effects of salidroside on LPS-induced HMGB1 release and then explored the underlying molecular mechanisms. We found that salidroside significantly inhibited LPS-induced HMGB1 release, and the inhibitory effect was correlated with the HMGB1 acetylation levels. Mechanismly, salidroside inhibits HMGB1 acetylation through the AMPK-SirT1 pathway. In addition, SirT1 overexpression attenuated LPS-induced HMGB1 acetylation and nucleocytoplasmic translocation. Furthermore, in SirT1 shRNA plasmid-transfected cells, salidroside treatment enhanced SirT1 expression and reduced LPS-activated HMGB1 acetylation and nucleocytoplasmic translocation. Collectively, these results demonstrated that salidroside might reduce HMGB1 release through the AMPK-SirT1 signalling pathway and suppress HMGB1 acetylation and nucleocytoplasmic translocation.

Effect of electroacupuncture at “Zúsānlľ” ( ST 36) and other lower he-sea points on the expression of TNF-α and NF-κB of acute gastric mucosal lesion rats

ObjectiveTo contrastively observe the effect of electroacupuncture at lower he-sea points of stomach, large intestine, small intestine and gallbladder of acute gastric mucosal lesion (AGML) model rats on the expression of serum high mobility group protein B 1 (HMGB 1), tumor necrosis factor-α (TNF-α) and nuclear factor-κB (NF-κB) of gastric tissue, and to explore whether the lower he-sea points of stomach have relative specificity in treatment of stomach diseases. MethodsSeventy healthy SD rats were randomly divided into seven groups, including control group, model group, “Zúsānlľ” (▪ ST 36) group, “Shàngjùxū” (▪ ST 37) group, “Xiàjùxū” (▪ ST 39) group, “Yánglíngquán” (▪. GB 34) group and “Chéngjīn” (▪ BL 56) group, with five male rats and five female rats in each group. Acute gastric mucosal lesion model was established by applying WRS method. After successful modeling and treatment for ten days, sampling was conducted, gastric mucosal lesion was observed visually and scored, the levels of serum HMGB 1 and gastric tissue TNF-α were detected by ELISA method, and the expression of NF-κB was detected by immunohistochemical method. Results▪ Compared with the model group, the levels of Ulcers and inflammation (Ul) in gastric mucosal and serum HMGB1, and the expression of gastric tissue TNF-α and NF-κB in the four lower he-sea points groups reduced (P<0.01 or P<0.05); (2) compared with ST 36 group, the levels of gastric mucosal Ul and serum HMGB1, and the expression of gastric mucosal TNF-α and NF-κB in other treatment groups increased significantly, and the difference was statistically significant (P<0.01 or P<0.05). Conclusion▪ Electroacupuncture at relevant lower he-sea points can inhibit the body's immune response, relieve inflammatory reaction and reduce mucosal lesion by regulating HMGB 1, TNF-α and NF-κB, so as to realize the intervention and treatment on AGML rats; ▪ the total intervention effect in ST 36 group is superior to that in other groups, indicating partially that there is a relative specificity between ST 36 and stomach.

TIMING OF TRAUMATIC BRAIN INJURY AND ITS SYSTEMIC EFFECTS ON THE LUNGS OF ADULT MALE ALBINO RATS

Objective: Timing of traumatic brain injury (TBI) is very common and important especially, in forensic autopsy cases with no signs of intracranial hemorrhage or contusion and with short time of survival following head injury. In such cases it is difficult to determine the cause of death. TBI results in systemic inflammatory responses that are complicated by dysfunction of peripheral organs such as the lungs, liver and intestine. The lung is especially critical in the setting of lung transplantation, where more than half of donor allografts are obtained postmortem from individuals with TBI. The aim of the present study was to assess post-intervals of traumatic brain injury and to evaluate the effects of TBI on lungs by measuring serum levels of High mobility group box 1 protein (HMGB1), performing quantitative real-time polymerase chain reaction (qPCR) analysis for gene expression and detecting any histopathological changes or immunohistochemical reaction of brain and lungs. Materials and Methods: Fifty four adult male albino rats divided into in to 3 groups (18 rats each): Group I: Negative control: rats left without any intervention. Group II: (sham): rats were anesthetized with ether inhalation and left without TBI inflection. Group III (TBI): rats were anesthetized with ether inhalation then the head was placed on platform and brain injury was done by weight drop method. After TBI infliction, six rats from group III (TBI) at each time point (1 st, 3rd& 7thday) and six rats from group I&II were anesthetized and blood samples were drawn for measuring serum level of (HMGB1).Then rats were sacrificed, specimens from contused cerebral cortex and lungs were subjected to quantitative real-time polymerase chain reaction (qPCR) analysis for gene expression of HMCB1, histopathology examination and immunohistochemical detection of HMGB1 immunoreaction. Results: Traumatic brain injury induced time dependent up-regulation of HMGB1 in serum and qPCR gene expression. Significant histopathological inflammatory changes and positive HMGB1 immunoreaction of brain and lung tissues of TBI (group III) were detected when compared with both control groups. Conclusion: It can be concluded that traumatic brain injury induced time dependent changes in serum and tissue levels of HMGB1, also histopathological changes in the brain and lungs of adult male albino rats. So, this marker can be useful for the diagnosis and timing of brain trauma and predict post TBI lung injury.

TLR4-Activated MAPK-IL-6 Axis Regulates Vascular Smooth Muscle Cell Function

Migration of vascular smooth muscle cells (VSMCs) into the intima is considered to be a vital event in the pathophysiology of atherosclerosis. Despite substantial evidence supporting the pathogenic role of Toll-like receptor 4 (TLR4) in the progression of atherogenesis, its function in the regulation of VSMC migration remains unclear. The goal of the present study was to elucidate the mechanism by which TLR4 regulates VSMC migration. Inhibitor experiments revealed that TLR4-induced IL-6 secretion and VSMC migration were mediated via the concerted actions of MyD88 and TRIF on the activation of p38 MAPK and ERK1/2 signaling. Neutralizing anti-IL-6 antibodies abrogated TLR4-driven VSMC migration and F-actin polymerization. Blockade of p38 MAPK or ERK1/2 signaling cascade inhibited TLR4 agonist-mediated activation of cAMP response element binding protein (CREB). Moreover, siRNA-mediated suppression of CREB production repressed TLR4-induced IL-6 production and VSMC migration. Rac-1 inhibitor suppressed TLR4-driven VSMC migration but not IL-6 production. Importantly, the serum level of IL-6 and TLR4 endogenous ligand HMGB1 was significantly higher in patients with coronary artery diseases (CAD) than in healthy subjects. Serum HMGB1 level was positively correlated with serum IL-6 level in CAD patients. The expression of both HMGB1 and IL-6 was clearly detected in the atherosclerotic tissue of the CAD patients. Additionally, there was a positive association between p-CREB and HMGB1 in mouse atherosclerotic tissue. Based on our findings, we concluded that, upon ligand binding, TLR4 activates p38 MAPK and ERK1/2 signaling through MyD88 and TRIF in VSMCs. These signaling pathways subsequently coordinate an additive augmentation of CREB-driven IL-6 production, which in turn triggers Rac-1-mediated actin cytoskeleton to promote VSMC migration.

Serum levels of HMGB1 have a diagnostic role in metastatic renal cell cancer.

RCC constitutes approximately 90% of all renal malignancies and 2-3% of all malignant tumours in adults. In spite of the improvement in radiologic methods, nearly 30% of the early metastatic RCC patients are incidentally diagnosed. HMGB1 is an extracellular signalling molecule that plays a role both in inflammation and carcinogenesis. Patients who were followed in Medical Oncology Departments of Denizli Government Hospital and Antalya Education and Research Hospital with a histopathological diagnosis of RCC between years 2010-2012 were enrolled in this study. HMGB1 levels were also assessed in a manually performed quantitative sandwich-enzyme-linked immunosorbent assay (ELISA) assay kit. In our study, we showed that the serum level of HMGB1, whether 149.9 pg/ml or not is important in differential diagnosis between patient and control group.

The damage-associated molecular pattern HMGB1 is elevated in human alcoholic hepatitis, but does not seem to be a primary driver of inflammation.

The role of sterile inflammation caused by release of damage-associated molecular patterns (DAMP) remains unclear in human alcoholic hepatitis (AH). The DAMP, high mobility group box-1 protein (HMGB1) is released by tissue damage and inflammation. We aimed to investigate whether HMGB1 is a primary inflammatory driver in AH by determining HMGB1 serum levels and effects on inflammatory cells from AH patients. We measured serum HMGB1 in 34 AH patients and 10 healthy controls using ELISA. Toll-like receptor 4 (TLR4) and CD14 expressions were assessed by flow cytometry on HMGB1-stimulated peripheral blood mononuclear cells (PBMC) and ELISA was used to measure TNF-α and IL-1β in the supernatants. We observed 5-fold higher serum levels of HMGB1 in AH patients at the day of diagnosis and day 30, but no associations to clinical outcome. HMGB1 stimulation increased the expression of TLR4 on CD14+-monocytes compared with unstimulated cells in the AH patients. The TNF-α and IL-1β production in response to HMGB1 was diminished in AH patients. In conclusion, AH patients have increased levels of HMGB1 in their blood. This combined with an increased TLR4 expression, but an unaffected cytokine response to HMGB1 suggest that HMGB1 is not the primary driver of inflammation in AH.