Improvement effects of glycyrrhizin on Helicobacter pylori-associated gastritis in rats and its mechanism / 中国药房

OBJECTIVE To investigate the improvement effects of glycyrrhizin （GL） on Helicobacter pylori （HP）-associated gastritis in rats and its mechanism. METHODS HP-associated gastritis rat model was induced by inoculating with 1×109 cfu/mL HP. The model rats were randomly divided into model group， positive control group （HP standard quadruple group）， GL low-dose， medium-dose and high-dose groups （5， 20， 50 mg/kg）， with 12 rats in each group. Another 12 healthy rats were selected as normal control group. Except the normal control group and model group were given constant volume of normal saline intragastrically， the other groups were given corresponding drugs intragastrically， once a day， for 30 consecutive days. After administration， rats received 13C urea breath test， and delta-over-baseline （DOB） was recorded； the pathological and cellular morphological changes of gastric mucosa in rats were observed， and pathological scoring was performed； the levels of interleukin-8 （IL-8）， IL-1β， tumor necrosis factor-α （TNF-α）， reactive oxygen species （ROS） and malondialdehyde （MDA） were detected in gastric mucosa of rats； mRNA expressions of high mobility group box-1 protein （HMGB1） and nuclear factor-κ-B （NF-κB）， relative expressions of nitric oxide synthases （iNOS） and HMGB1， the phosphorylation level of NF- κBp65 were also detected in rats. RESULTS Compared with normal control group， the DOB value， histopathological score of gastric mucosa， the levels of IL-8， IL-1β， TNF-α， ROS and MDA， relative expressions of HMGB1 and NF- κB mRNA， relative expressions of iNOS and HMGB1 protein and the phosphorylation level of NF-κB p65 were all increased significantly in model group （P＜0.05）； the epithelial cells of gastric mucosa in rats were incomplete in structure and decreased in the number， with an increase in cell fragments and vacuoles， and significant cell pyknosis. Compared with model group， the changes of the above indexes in GL groups and positive control group were significantly reversed （P＜0.05）； the changes in the above indicators in the GL high-dose group were more significant than GL low-dose and medium-dose groups （P＜0.05）； the pathological changes of gastric mucosal cells in rats had all improved. CONCLUSIONS GL may inhibit inflammation and oxidative stress by inhibiting the activation of HMGB1/NF-κB pathway， thus relieving HP-induced gastric mucosal injury.

Effect of Modified Erchentang on Bronchioles of Rats with Chronic Obstructive Pulmonary Disease by HMGB1/RAGE/NF-κB Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo study the effect of modified Erchentang on the expression of key molecules in the high mobility group Box 1 protein （HMGB1）/receptor for advanced glycation endproduct （RAGE）/nuclear factor-κB （NF-κB） signaling pathway in bronchioles of rats with chronic obstructive pulmonary disease （COPD）， to explore the mechanism of modified Erchentang against bronchiolar inflammation of COPD rats via HMGB1/RAGE/NF-κB signaling pathway. MethodSixty SD rats were randomly divided into normal group， model group， modified Erchentang low-， medium- and high-dose groups （5， 10， 20 g·kg-1·d-1） and ethyl pyruvate （HMGB1 inhibitor） group， with 10 in each group. The COPD rat model was prepared by cigarette smoke combined with tracheal injection of lipopolysaccharide （LPS）. After modeling， the modified Erchentang groups were given corresponding drugs （ig） and Ringer's solution （4 mL， ip）， while the EP group was treated with equal volume of normal saline （ig） and EP （0.04 g·kg-1·d-1， ip）. The normal group and the model group received equal volume of normal saline （ig） and Ringer's solution （ip） for 21 consecutive days. The contents of HMGB1， chemokine （C-X-C motif） ligand 1 （CXCL1）， CXCL2 and monocyte chemotactic protein-1 （MCP-1） in bronchoalveolar lavage fluid （BALF） were detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA expressions of HMGB1， RAGE and NF-κB p65 were determined by Real-time polymerase chain reaction （Real-time PCR）， and the protein expressions of HMGB1， RAGE， p-NF-κB p65， and alpha-smooth muscle actin （α-SMA） in bronchioles tissue of rats were determined by immunohistochemistry （IHC）. ResultCompared with the conditions in the normal group， the forced vital capacity （FVC）， forced expiratory volume in the first second （FEV1） and FEV1/FVC in the model group were decreased （P<0.01） while the contents of HMGB1， CXCL1， CXCL2 and MCP-1 in BALF were increased （P<0.01）. And the model group presented higher mRNA expressions of HMGB1， RAGE and NF-κB p65 （P<0.01） and protein expressions of HMGB1， RAGE， p-NF-κB p65 and α-SMA （P<0.05， P<0.01） than the normal group. Compared with the model group， the modified Erchentang medium- and high-dose groups had increased FEV1/FVC （P<0.05， P<0.01）， lowered contents of HMGB1， CXCL1， CXCL2 and MCP-1 in BALF （P<0.05， P<0.05）， and reduced mRNA expressions of HMGB1， RAGE and NF-κB p65 （P<0.05， P<0.01） and protein expressions of HMGB1， RAGE， p-NF-κB p65 and α-SMA （P<0.05， P<0.01）. ConclusionModified Erchentang can resist bronchiolar inflammation of COPD rats. The mechanism may be related to down-regulating the mRNA expressiona of HMGB1 and RAGE， inhibiting the activity of NF-κB， and reducing the release of HMGB1， CXCL1， CXCL2 and MCP-1， thus suppressing the inflammatory injury and abnormal repair of bronchioles.

miR-142-3p alleviates house dust mite induced airway inflammation among children / 中国学校卫生

Objective@#To investigate the role of miR-142-3p in alleviation of house dust mite induced allergic airway inflammation among children, so as to provide insights into unraveling the pathogenesis of allergic airway inflammation.@*Methods@#Serum samples were collected from 15 patients with house dust mite induced allergic asthma and 15 healthy children in Jiangnan University Medical Center from September to November 2022, and serum miR-142-3p expression was quantified using a fluorescent quantitative real time PCR (qPCR) assay. The levels of interleukin 6 (IL 6) and tumor necrosis factor α (TNF α) were measured in the cell culture supernatant using enzyme linked immunosorbent assay (ELISA), and the expression of high mobility group box 1 (HMGB1) was detected at transcriptional and translational lvels using qPCR and Western blotting assays. The negative regulation of the HMGB1 gene by miR 142 3p was identified using a dual luciferase gene reporter assay, and the expression of downstream regulatory proteins was determined in human normal lung epithelial cells (BEAS 2B) cells transfected with miR 142 3p using Western blotting. In addition, female C57BL/6 mice at ages of 6-8 weeks were randomly assigned to the phosphate buffer saline (PBS) group, house dust mite sensitized airway inflammation group and house dust mite sensitized airway inflammation + miR 142 3p intervention group. Mouse airway inflammation was evaluated using hematoxylin eosin staining, and the expression of inflammatory cells and inflammatory cytokines were detected in mouse bronchoalveolar lavage fluid (BALF) using Giemsa staining and ELISA.@*Results@#Lower serum miR-142-3p expression was quantified among children with house dust mite induced allergic asthma than among healthy controls (1.33±0.21 vs. 4.74±0.62, t=5.22, P <0.05). Stimulation with dermatophagoides farinae extract (DFE) resulted in a reduction in miR-142-3p expression in BEAS-2B cells (0.82±0.25), while transfection with miR-142-3p mimics resulted in a rise in miR-142-3p expression in BEAS-2B cells (0.55±0.14)( t=3.31, 3.94, P <0.05). Pre treatment with miR-142-3p reduced the expression of IL 6(2.25±0.46)and TNF α(6.58±1.95) ( t=4.86, 3.38, P <0.05) in BEAS 2B cells stimulated with DFE, and treatment with miR-142-3p mimics resulted in a reduction in TLR4 and NF-κB expression in BEAS-2B cells via negative regulation of the HMGB1 expression. In addition, treatment with miR-142-3p was found to alleviate inflammatory cell infiltration in lung tissues of house dust mite sensitized mice, and results in a reduction in interleukin 4 (IL-4)[(107.60±10.43)pg/mL], interleukin 5 (IL 5)[(95.78±13.14)pg/mL] and HMGB1[(2.52±0.87)pg/mL] expression in BALF ( t=10.32, 7.29, 2.90, P <0.05).@*Conclusion@#miR-142-3p alleviates house dust mite induced allergic airway inflammation among children via negative regulation of the HMGB1/TLR4/NF-κB pathway.

Effects of dexamethasone on neutrophil airway inflammation and HMGB1 expression in asthmatic mice / 解放军医学杂志

Objective: To investigate the effect of dexamethasone intervention on neutrophil airway inflammation and high mobility group box 1 protein (HMGB1) expression in asthmatic mice. Methods: Forty healthy SPF grade female Balb/c mice were randomly divided into four groups (n=10): Control group, Asthma group, 1 mg/kg dexamethasone intervention group and 5 mg/kg dexamethasone intervention group. The asthmatic mice model was induced by egg-free ovalbumin. The airway hyper-responsiveness was measured by the invasive pulmonary function instrument; the number and classification of inflammatory cells in bronchoalveolar lavage fluid (BALF) were detected; lung pathological changes were observed by HE staining; HMGB1 expression was detected by immunohistochemistry and Western blotting. Results: Compared with Control group, the airway responsiveness of asthmatic mice was significantly enhanced (P0.05). Conclusions: The expression of HMGB1 in airway epithelium and lung tissue of asthmatic mice with neutrophilic airway inflammation was significantly increased. After dexamethasone intervention, asthmatic mice still showed higher neutrophilic airway inflammation and expression of HMGB1 in lung tissue, suggesting that HMGB1 may be associated with neutrophilic airway inflammation and glucocorticoid resistance in asthma.

The effect of serum HMGB1 levels on the severity and prognosis of critical ill patients with early trauma-induced coagulopathy in the intensive care unit / 解放军医学杂志

Objective: To analyze the influence of serum high mobility group box-1 protein (HMGB1) levels on the severity and prognosis of critical ill patients at the early of trauma-induced coagulopathy (TIC) in intensive care unit (ICU). Methods: 43 cases of critical ill patients with severe trauma were included during January 1, 2017 to December 31, 2018 in ICU of Foshan Hospital of Traditional Chinese Medicine. International normalized ratio (INR) >1.2 was applied as the diagnosis criterion of TIC. The patients was divided into TIC group (n=23) and control group (n=20). Their age, sex, injury mechanism, the interval between injury and admission to ICU (delay time), the interval between injury and coagulopathy correction (correction time), ISS scores, APACHE II scores at admission to ICU were recorded, and the activated partial thromboplastin time (APTT), prothrombin time (PT), INR, fibrinogen (Fib), platelet counts (PLT), C-reactive protein (CRP) and procalcitonin (PCT) levels were detected meanwhile. The serum HMGB1 levels were examined through ELISA. The blood transfusion volume (red blood cells, RBC and fresh frozen plasma, FFP), ventilation time, ICU stay and 28-day mortality rate were statistically analyzed. Results: TIC occurred in 53.5% of critical ill trauma patients in ICU. There were no significant differences in the age, sex, injury mechanism, delay time, APACHE II scores, ISS scores, APTT, PT, CRP and PCT levels between two groups (P>0.05). Compared with control group, the Fib and PLT levels were significantly reduced in the TIC group, and the ventilation time, blood transfusion volume of RBC and FFP, infection rate and organ dysfunction rate were remarkably increased (P<0.05). Besides, the 28-day mortality rate revealed a raised tendency in TIC group (P=0.091). Simultaneously, the serum HMGB1 levels at admission to ICU were significantly increased in the TIC group, and the serum HMGB1 level in the death subgroup was much higher than that in the survivors, as same as those in the TIC subgroup analysis (P=0.000). The correlation analysis disclosed that serum HMGB1 levels at admission was positively related to delay time, correction time, APACHE II score, ISS score, ventilation time, INR levels, APTT, CRP and PCT levels at admission (r=0.648, 0754, 0.526, 0.516, 0.521, 0.509, 0.432, 0.592, 0.375), and was negatively associated with Fib levels, PLT value, infection incidence rate, organ dysfunction rate and 28-day mortality rate (r=-0.424, -0.571, -0.505, -0.396, -0.765). There were significant differences in the delay time, correction time, ISS scores, transfusion volume, serum HMGB1 levels, PLT value, APTT and CRP levels between death subgroup and survivor subgroup of TIC patients (P<0.05), and serum HMGB1 levels and PLT value at admission were independent risk factors in the multivariable logistic regression analysis (P=0.004, 0.011). For the TIC sub-group trauma patients, the AUC of serum HMGB1 levels was 0.897(95%CI 0.748-1.000, P=0.002), the best cutoff value was 54.60 ng/ml with Youden index of 0.808. Conclusions: The PLT levels and serum HMGB1 levels at admission to ICU are independent risk factors of critical ill patients with severe trauma. The serum HMGB1 levels is closely related to severity and prognosis, and can predict the 28-day mortality rate of critical ill patients with TIC.

Anti-inflammatory effect of impressic acid from Acanthopanax gracilistylus based on NF-κB signaling pathway / 中草药

Objective: To explore the anti-inflammatory effect of impressic acid (IA) from Acanthopanax gracilistylus on lipopolysaccharide (LPS)-induced RAW264.7 cells. Methods: RAW264.7 cells were stimulated by LPS to establish an inflammatory model. The cytotoxicity of IA on RAW 264.7 cells was detected by EZ4U cell proliferation and cytotoxicity analysis kit. The level of nitric oxide (NO) was determined by Griess. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were determined by ELISA. The expressions of TNF-α and IL-1β mRNA were detected by RT-PCR. The expression of high-mobility group box 1 protein (HMGB1) was detected by western blotting. The levels of nuclear factor-κB (NF-κB) in cytoplasm and nucleus were measured by ELISA. Results: IA significantly suppressed the levels of TNF-α and IL-1β, the expression of HMGB1 protein, and the translocation of NF-κB from cytoplasm to nucleus in LPS-induced RAW264.7 cells (P < 0.05, 0.01). Conclusion IA from A. gracilistylus has an anti-inflammatory effect on LPS-induced RAW264.7 cells.

Regulatory role of high mobility group box-1 protein in the balance of Th17/Treg in peripheral blood of immune thrombocytopenia patients / 白血病·淋巴瘤

Objective@#To explore the effect of high mobility group box-1 protein (HMGB1) on the balance of Th17/Treg in patients with immune thrombocytopenia (ITP).@*Methods@#A total of 30 patients who were first diagnosed as ITP in the Fifth People's Hospital of Datong from July 2017 to April 2018 were selected as the case group, and another 30 healthy volunteers in the corresponding period were taken as the control group. The proportion of Th17 and Treg cells was detected by using flow cytometry, and the concentration of HMGB1, interleukin (IL)-17 and transforming growth factor β (TGF-β) in plasma was tested by using enzyme-linked immunosorbent assay (ELISA). Isolated peripheral blood mononuclear cells (PBMC) were cultured in vitro. After the treatment with recombinant human HMGB1 (rhHMGB1), real-time polymerase chain reaction (RT-PCR) was applied to detect the mRNA expression changes in Treg cell transcription factor intracellular forkhead helix transcription factor 3 (Foxp3) and Th17 cell transcription factor retinoid related orphan receptor γt (RORγt). The differences of indicators in Treg cell transcription factor peripheral blood between the case group and the control group were compared, and the balance correlation between HMGB1 and Th17/Treg was analyzed.@*Results@#Compared with the healthy control group, the proportion of Th17 cells and the expression level of HMGB1 and IL-17 in peripheral blood of ITP patients were increased (all P < 0.01), while the proportion of Treg cells and the level of TGF-β were decreased (all P < 0.01). The proportion of Th17 cells and the expression level of IL-17 and HMGB1 in peripheral blood of ITP patients were positively correlated with the concentration of HMGB1 (all P < 0.01); the proportion of Treg cells and the level of TGF-β were negatively correlated with the expression level of HMGB1 (all P < 0.01). In vitro experiments, the expression of intracellular RORγt mRNA was increased compared with the negative control group (1.50±0.24 vs. 0.93±0.22, t = 9.612, P < 0.01), and the expression of Foxp3 mRNA was decreased compared with the negative control group after the stimulation of PBMC by rhHMGB1 (0.72±0.19 vs. 1.08±0.18, t = 7.387, P < 0.01).@*Conclusion@#The high level of HMGB1 in the peripheral blood of ITP patients induces Th17/Treg imbalance and aggravates inflammatory reactions, which may be an important cause of ITP.

Effects of Dexmedetomidine Combined with Mild Hypothermia on the Levels of HMGB 1，TLR4 and Trem- 1 in Lung Tissues of Sepsis Model Rats / 中国药房

OBJECTIVE： To investigate the effects of dexmedetomidine combined with mild hypothermia on the levels of High mobility group box 1 protein （HMGB1）， Toll like receptor 4 （TLR4） and Triggering receptor expressed on myeloid cells 1 （Trem-1） in lung tissues of sepsis model rats. METHODS： Totally 100 SD rats were randomly divided into sham operation group （normal saline）， model group （normal saline）， dexmedetomidine group （2 μg/kg）， mild hypothermia group （normal saline+anal temperature 32-35 ℃ caused by whole body spraying of cold water） and combination group （dexmedetomidine 2 μg/kg+anal temperature 32-35 ℃ caused by whole body spraying of cold water）， with 20 rats in each group. Except that sham operation group received sham operation， sepsis model was induced in other groups. After ligation and incision， the corresponding drugs were pumped into the jugular vein and the corresponding body temperature was maintained. Plasma samples were collected 6 h after operation. Interleukin 1 （IL-1）， IL-6， tumor necrosis factor α （TNF-α） were determined by ELISA. The lung wet mass/dry mass ratio （W/D） was calculated by weighing the mass. Lung tissue sections were observed by HE staining， and lung injury scores were scored. The activity of MPO in lung tissue was determined by immunoturbidimetry. The expression of HMGB1，TLR4 and Trem-1 protein was determined by Western blot. RESULTS： Compared with sham operation group， the contents of IL-1， IL-6 and TNF-α， W/D， lung injury score， MPO activity， the protein expression of HMGB1， TLR4 and Trem-1 were increased significantly， with statistical significance （P＜0.05）； lung tissue section showed that alveolar wall was obviously thickened； a large number of inflammatory cells infiltrated； blood vessels were obviously dilated and congested. Compared with model group， above indexes of rats in dexmedetomidine group， mild hypothermia group and combination group were decreased significantly， with statistical significance （P＜0.05）； the degree of pathological changes in lung tissue was significantly reduced. Compared with dexmedetomidine group and mild hypothermia group， above indexes of combination group were decreased more significantly， with statistical significance （P＜0.05）. Alveolar walls were thickened， inflammatory cell infiltration was relieved significantly and no vascular diatation and congestion was found. CONCLUSIONS： Dexmedetomidine combined with mild hypothermia can significantly improve lung injury in sepsis model rats， and down-regulate the protein expression of HMGB1， TLR4 and Trem-1. Therapeutic efficacy of combination therapy is better than single therapy.

Influence of high mobility group box-1 protein inhibition on protein expression of rage/toll-like receptor 4 signal passway in ht22 cells induced with aβ25-35 / 中国药理学与毒理学杂志

OBJECTIVE To investigate the influence of high mobility group box-1 protein (HMGB1) inhibition on the expression of receptors for advanced glycation end products (RAGE)/Toll-like receptor 4 (TLR4) signal passway related proteins, RAGE, TLR4, NF-kB, interieukin-1β (IL-1βand tumor necrosis factor-α(TNF-α) in HT22 cells stimulated with amyloid-β25-35 (Aβ25-35). METHODS HT22 cells were stimulated with Aβ25-350, 2.5, 5, 10, 20 and 40 ymol • L"1 for 24 h, and the vitality of HT22 cells was determined using MTT assay. Half maximal inhibitory concentration (IC50) was calculated. HT22 cells were divided into 5 groups: Normal cell control group, Apas-K 40 Mmol-L"' group, siRNA 50 Mmol-L"' group, siRNA or scramble siRNA+APas-as group (HT22 cells were transfected with siRNA or scramble siRNA at 50 pmol • L"' for 24 h, then treated with synthetic Apa-as at a final concentration of 40 pmol • L"1 for 24 h). The morphology of HT22 cells was observed under a microscope. The location of HMGB1 was observed by immunofluorescence. The protein expressions of HMGB1, RAGE, TLR4 and NF-kB P65 were detected by Western blotting. The levels of IL-IP and TNF-α in the culture supernatants were examined by ELISA. RESULTS The ICso of HT22 cells treated with Apas-as for 24 h was 41.17 Mmol-L-1, and A25-35 40 Mmol-L"1 was selected in subsequent experiments. After 24 h treatment with A25-35 40 nmol-L"', a large number of cells died, or aggregated into clusters, processes decreased, cell gaps increased, and HMGB1 was released from the nucleus to the outside. The protein expressions of HMGB1, RAGE, TLR4 and NF-kB P65 in HT22 cells were significantly increased (P<0.05) after treatment with Apas-as, and the levels of IL-1p and TNF-α in the culture supernatants were significantly increased (P<0.05). The protein expressions of HMGB1, RAGE, TLR4 and NF-kB P65 in HT22 cells were significantly decreased (P<0.05) after 24 h treatment with siRNA 50 pmol-L"' and the levels of IL-1p and TNF-α in the culture supernatants were significantly decreased (P<0.05). CONCLUSION RNA interference with HMGB1 reduces the expression of HMGB1, RAGE, TLR4 and NF-kB P65 in HT22 cells stimulated with A25-35

San-Cao Granule () Ameliorates Hepatic Fibrosis through High Mobility Group Box-1 Protein/Smad Signaling Pathway / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the possible mechanism of San-Cao Granule (SCG, ) mediating antiliver fibrosis.</p><p><b>METHODS</b>A total of 60 male Sprague-Dawley rats were randomly divided into the normal control group, porcine serum-treated group, ursodesoxycholic acid (UDCA, 60 mg/kg), SCG (3.6 g/kg) group, SCG (1.8 g/kg) group and SCG (0.9 g/kg) group, with 10 rats in each group. Liver fibrosis was induced with porcine serum by intraperitoneal injection for 8 weeks, except for the normal control group. Then, the rats in the three SCG-treated groups and UDCA group were administered SCG and UDCA respectively for 4 weeks. The serum levels of alanine transaminase (ALT), aspartate transaminase (AST), albumin (ALB), total bilirubin (TBIL), hyaluronic acid (HA), laminin (LN), and type IV collagen (IVC) were examined using commercial kits and hepatic histopathology was examined with hematoxylin and eosin and Masson staining. Moreover, the protein expression levels of high mobility group box-1 protein (HMGB1), transforming growth factor β1 (TGF-β1), phosphorylated mothers against decapentaplegic homolog 3 (p-Smad3), Smad7, toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), nuclear factor-kappa B (NF-κB) and α-smooth muscle actin (α-SMA) were determined by western blot, immunohistochemistry and real time quantitative-reverse transcription polymerase.</p><p><b>RESULTS</b>Both SCG (3.6 and 1.8 g/kg) and UDCA significantly ameliorated the liver fibrosis induced by porcine serum as indicated by retarding the serum levels increasing of ALT, AST, TBIL, HA, LN and IVC and preventing the serum level reducing of ALB compared with the model group (all P<0.01). Meanwhile, the collagen deposition was attenuated by SCG and UDCA treatment. Furthermore, SCG markedly reduced the expressions of HMGB1, TGF-β1, p-Smad3, TLR4, MyD88, NF-κB and α-SMA, and enhanced the expression of the Smad7 compared with the model group (all P<0.01).</p><p><b>CONCLUSION</b>SCG ameliorates hepatic fibrosis possibly through inhibiting HMGB1, TLR4/NF-κB and TGF-β1/Smad signaling pathway.</p>

The immune response in acute and chronic pancreatitis / 解放军医学杂志

Acute and chronic pancreatitis are most common gastrointestinal diseases.Recently,there are emerging evidences that immune cell play important roles in the pathogenesis of acute and chronic pancreatitis.Studies have shown that macrophages,high mobility group protein 1 (HMGB 1) and interleukin-33 (IL-33) play an important role in the pathological process of pancreatitis,and are regulated by multiple levels.For example,immune cells are critical in the development and progression of pancreatitis,which not only have the ability to induce microenvironment,but also respond to danger signals derived from endogenous and exogenous molecules.Therefore,further understanding of relevant immune signaling will provide new idea and potential therapeutic targets that can prevent disease progression.Here,we review recent data from animal and human clinical studies that focus on immune responses.

Advances in the study of NETs in the pathogenesis of chronic obstructive pulmonary disease / 医学研究生学报

Chronic obstructive pulmonary disease (COPD) is a prevalent chronic inflammatory disease of the lung in the worldwide. With the deeper development of the study, people gradually realized that COPD also with the characteristics of autoimmune diseases. However, the initiation mechanism of acquired immunity in COPD is still unclear. Chronic neutrophilic inflammation of the airways is a distinct feature of COPD. The latest research shows that neutrophils can form a reticular substance composed of DNA in the infected state-NETs, which can not only give full play to the anti infection effect, but also cause tissue damage. In some autoimmune diseases, it can even initiate the acquired immune responses. This paper will briefly review the recent advances of NETs in the pathogenesis of COPD and its potential role as an anti-inflammatory target for COPD, so as to provide new ideas for the anti-inflammatory treatment of COPD in the future.

Study on the Effects of HMGB1-mediated Inflammatory Pathway on Liver Injury Induced by Tripterygium wilfordii / 中国药房

OBJECTIVE:To study the effects of high-mobility group box 1 protein(HMGB1) -mediated inflammatory pathway HMGB1-Toll like receptor 4 (TLR4) /nuclear transcription factor κ B(NF-κ B)on liver injury of rats induced by Tripterygium wilfordii,and to provide reference for clarify the mechanism of liver injury induced by T.wilfordii. METHODS:Totally 24 SD rats were randomly divided into blank group(normal saline,i. g. ),T. wilfordii group(16 g/kg by crude drug,i. g. )and neutralizer group(16 g/kg T. wilfordii crude drug i. g. after i. p injection of 100 mg/kg Ammonium glycyrrhizinate solution 3 h),with 8 rats in each group. All rats were treated for consecutive 3 weeks. The serum levels of AST and ALT in rats were detected every week. After the end of medication,the serum levels of HMGB1,IL-1β,IL-2 and TNF-α were detected by ELISA method;the protein expression of HMGB1,NF-κB p65 and TLR4 in liver tissue of rats were detected by Western blot assay. The pathological changes of liver tissue in rats were measured with HE staining method. RESULTS:After 3 weeks of treatment,the serum levels of AST, ALT,HMGB1,IL-1β,IL-2 and TNF-α in rats,the protein expression of HMGB1,NF-κB p65 and TLR4 in liver tissue of rats in T. wilfordii group were significantly higher than blank group and neutralizer group(P<0. 05 or P<0. 01). Hepatocyte edema was found around the central vein of the liver,and circular vacuoles were seen in some hepatic cytoplasm in T. wilfordii group;only varying size of vacuoles were found in a small number of cells in neutralizer group. CONCLUSIONS:T. wilfordii induced liver injury may be associated with the activation of HMGB1-TLR4/NF-κB inflammation pathway.

Research progress of high mobility group box 1 in pediatric diseases / 中国小儿急救医学

High mobility group box 1 protein(HMGB1) is widely present in eukaryotic nuclei and named for its rapid migration during polyacrylamide gel electrophoresis. Studies have shown that HMGB1 playes an important role in the development of gene transcription,inflammation,cancer and other diseases. In this paper, the biological characteristics of HMGB1 and its research progress in pediatric diseases are reviewed.

The role of RAGE in high mobility group box-1 protein1-mediated CD4+T cells differentiation to Th1/Th2 / 中华急诊医学杂志

Objective To explore the role of receptor for advanced glycation end products (RAGE) in HMGBl-mediated CD4+ T cells differentiation to Th1/Th2.Methods CD4+ T lymphocytes isolated from the spleens of male BALB/C mice by magnetic beads were suspended in RPMI-1640 with 10％ FCS in 2× 106cell/well on 96-well cell culture plates in vitro.The cells were randomly divided 4 groups according to concentration of HMGB1 treatment:control group,10 ng/mL group,100 ng/mL group,1 000 ng/mL group after stimulation with ConA in 3 μg/mL for 12 hours.IL-4 and IFN-γ levels in culture supernatants were quantitated by ELISA kits after HMGB1 stimulation for 12,24,and 48 h.According to the results,cultured cells were exposed to HMGB1 in 100 ng/mL for 24 h in the following experiments.The cells were randomly divided into 4 groups:control group,A group,B group,C group,and each group were cultured with ConA in 3 μg/mL for 24 h.The cells of control group and other three groups were stimulated with PBS or 100 μg/L HMGB1 for 24 h.The cells of B,C groups were incubated with 1/200 diluted 5 μg/L anti-RAGE Abs (anti-bodies) or PBS for 2 h before HMGB1 stimulation.The cell suspension was obtained to detect the levels of IL-4 and IFN-γ by EILSA and the protein levels and mRNA expressions of RAGE,CATA-3 were detected by western-blot and real-time fluorescent quantitative PCR,respectively.ResuRs Compared with control group,CD4+ T cells incubated with increasing concentrations of HMGB1 (100,1 000 ng/mL) for 24 h resulted in a decrease in IFN-γ/IL-4 ratio (P＜0.05).When CD4+ T cells were exposed to 100 ng/mL HMGB1 for 12 h,IFN-γ/IL-4 ratio was markedly increased.However,CD4+ T cells treated with 100 ng/mL HMGB1 for 24,48 h,IFN-γ/IL-4 ratio was markedly inhibited (P＜0.05).Compared with control group,protein levels and mRNA expressions of RAGE and GATA3 of cells in A group were significantly increased (P＜0.05),and IFN-γ/IL-4 ratio of cell suspension in A group and B group was significantly down-regulated (P＜0.05).Compared with A group,IFN-γ/IL-4 ratio of cell suspension in C group was increased (P＜0.05),and expression of GATA3 mRNA was down-regulated (P＜0.05).Compared with A group,protein level of RAGE of cells in C group was significantly down-regulated (P＜0.05),but protein level of RAGE of cells in C group was still increased compared with control group (P＜0.05).Conclusion Th1/Th2 differentiation induced by HMGB1 on CD4+ T lymphocytes in vitro was at least partly mediated by over-activating RAGE/GATA3 pathway.

High-mobility group box 1 protein (HMGB1) in maternal plasma affects Th17/Treg balance via receptor for advanced glycation end products( RAGE) -IL-6 pathway in preeclamptic pregnancies / 中华微生物学和免疫学杂志

Objective To analyze the relationships of high mobility group box 1 protein (HMGB1) with regulatory T cells (Treg), T helper 17 cells (Th17) and cytokine secrtion in peripheral blood of gravidas with preeclampsia(PE),and to investigate the mechanism of HMGB1 in regulating Th17/Treg ratio via receptor for advanced glycation end products (RAGE)-IL-6 pathway. Methods Forty gravi-das with mild(20 cases) and severe(20 cases) PE were recruited as experimental groups,20 heathy gravi-das in the third trimester of pregnancy were enrolled as control group. Concentrations of HMGB1,IL-6,IL-17 and TGF-β in peripheral blood of all subjects were determined by enzyme-linked immunosorbent assay (ELISA). Real-time quantitative PCR(RT-PCR) was used to detect the expression of RAGE at mRNA lev-el in peripheral blood mononuclear cells(PBMCs). The percentages of Treg and Th17 cells were determined by flow cytometry. RT-PCR was performed to analyze changes in the expression of RAGE,IL-6,Foxp3 and RORγt at mRNA level after the PBMCs isolated from 20 garvidas with PE were cultured in vitro and stimula-ted with recombinant human HMGB1 (rhHMGB1). Results The levels of HMGB1,IL-6,Th17 and IL-17 in peripheral blood of gravidas with PE were significantly higher than those in the normal pregnancy group. Moreover,HMGB1 level was positively correlated with IL-6 level and ratios of Th17/Treg and IL-17/TGF-β in preeclamptic pregancies. In vitro stimulation of PBMCs with rhHMGB1 significantly enhanced the expres-sion of RAGE,IL-6 and RORγt at mRNA level, but suppressed the expression of Foxp3 at mRNA level. Conclusion Enriched HMGB1 in plasma shifts the Th17/Treg balance towards Th17 dominance via the RAGE-IL-6 pathway, which exacerbates inflammation and participates in the onset of preeclampsia during pregnancy.

Expression of high mobility group box 1 protein in patients with ankylosing spondylitis / 中华风湿病学杂志

Objective To investigate the role of high mobility group box l protein (HMGB1) in the pathogenesis of ankylosing spondylitis (AS). Methods Enzyme-linked immuno sorbent assay (ELISA) was used to test the levels of plasma HMGB1 levels in 58 patients with active AS [bath ankylosing spondylitis disease activity index (BASDAI)>6, or 6>BASDAI>4 and erythrocyte sedimentation rate (ESR)>22 mm/1 h, 6>BASDAI>4 and hypersensitive C reactive protein (hsCRP)>9 mg/L], 73 cases of stable AS (BASDAI<4) and 70 healthy control. Twelve patients who were treated with TNF-alpha antagonist for 6 month were followed-up. Their plasma levels of HMGB1 were detected before and after treatment. Quantitative data were described by, while qualitative data were described by case number. Variance analysis or rank sum test was adopted for the difference between measurement data groups, LSD method was adopted for further pair-wise comparison. The correlation between variables was analyzed by using Spearman correlation analysis. Results The levels of plasma HMGB1, ESR, hsCRP, White blood cell WBC, GR, Mo and GLOB were significantly higher in the AS patients than those in the healthy control group (P<0.001), and the level of plasma HMGB1 in the AS patients was significantly positively correlated with BASDAI, Bath ankylosing spondylitis functional index (BASFI), ESR, hsCRP, WBC, GR, Mo, and GLOB (r=0.288, 0.174, 0.308, 0.243, 0.261, 0.301, 0.279, 0.289; P=0.004, 0.047, 0.000, 0.005, 0.003, 0.000, 0.001 ,0.001). The level of plasma HMGB1, BASDAI, BASFI, ESR, hsCRP, WBC, GR, GLOB were significantly higher in the active AS group than in the stable group (Z=-3.598,-9.456,-5.907, -2.562, -3.178, 4.134, -2.574, -4.582; P=0.000, 0.000, 0.000, 0.012, 0.002, 0.000, 0.011, 0.000). The level of plasma HMGB1 was not found statistically poor in the patients with different expressions of HLA-B27, or hip involvement and history of vuvitis (P>0.05). The plasma HMGB1 level, BASDAI, BAIFI, ESR, hsCRP and GLOB in the 12 followed-up patients were significantly decreased (P=0.034, 0.002, 0.002, 0.005, 0.004, 0.004) after being treated with biological agents for 6 months. Conclusion HMGB1 might play a vital role in the pathogenesis of ankylosing spondylitis,and the HMGB1 might be used as a clinical indicator to evaluate the activity of AS and to assess the clinical efficacy.

Mechanism of signal transduction in distant organ injuries induced by intestinal ischemia/reperfusion / 中华临床营养杂志

Objective To investigate the effect of different antibodies on Toll-like Receptor 4-High Mobility Group Box 1 and its downstream signal transductions in distant organ injuries caused by intestinal ischemia/reperfusion in mice.Methods A total of 40 mice (C57BL/6,SPF level) were by random number table method assigned into five groups:sham,control,anti-HMGB1,anti-Myeloid differentitation gene,and antiTIR domain containing adaptor inducing IFN-β (n=8).In the control,anti-HMGB1,anti-MyD88,and antiTRIF groups,the IgG,HMGB1,MyD88,and TRIF antibodies were injected,respectively,via the tail vein 30 minutes before ischemia (1 mg/kg body weight,0.025％).After anesthesia and abdomen incision,all mice,except the sham group,underwent intestinal ischemia by clamping the superior mesenteric artery for 60 minutes followed by 60 minutes of reperfusion.Sham group underwent the same surgical procedures except for clamping the artery.Serum nuclear factor-κB p65,Interleukin-6 and Tumor Necrosis Factor-α were measured.Morphological changes in the lung and intestine were evaluated.mRNA and protein expressions of HMGB1 and NF-κB in lung and intestinal tissues were assayed.Results Compared with the control group [(228.53± 24.85),(104.91±31.18),and (70.81±46.97) ng/L],HMGB1 [(145.00±33.63),(62.28±6.73),and (52.76± 5.71) ng/L],MyD88 [(191.12± 13.22),(85.90± 17.37),and (63.19 ± 5.47) ng/L],and TRIF [(183.73±10.81),(78.14±7.38),and (59.70±4.63) ng/L] significantly decreased the serum level of NF-κB (P=0.000,0.005,0.001),IL-6 (P=0.000,0.004,0.000) and TNF-α (P=0.000,0.024,0.002) after ischemia reperfusion.Tissue injuries in the lung and intestine were also alleviated by HMGB1,MyD88,and TRIF.The anti-HMGB1,anti-MyD88,and anti-TRIF groups displayed significant elevations of HMGB1 mRNA [lung (1.89±0.18),(2.35±0.31),and (2.29±0.28),ileum (4.93±0.55),(5.96± 0.73),and (5.76±0.51)],NF-κB mRNA [lung (1.42±0.23),(1.77±0.18) and (1.70±0.13),ileum (2.23±0.55),(3.11±0.38) and (2.99±0.24)] and NF-κB protein expressions in lung and ileum tissues compared to the sham group [lung HMGB1 mRNA (1.04±0.19) (P=0.000,0.000,0.000),NF-κBmRNA (1.03±0.21) (P=0.004,0.000,0.000),ileum HMGB1 mRNA (1.14±0.54) (P=0.000,0.000,0.000),NF-κB mRNA (1.03±0.23) (P=0.000,0.000,0.000)].However,incornparison with the control group [lung HMGB1 mRNA (2.67±0.23) (P=0.000,0.035,0.016),NF-κB mRNA (2.04±0.29) (P=0.000,0.039,0.012),ileum HMGB1 mRNA (6.70±0.66) (P=0.001,0.038,0.015),NF-κBmRNA (3.71±0.53) (P=0.000,0.018,0.006)],the other three groups showed a significant down-regulation,with the most remarkable decrement in the anti-HMGB1 group.Application of anti-HMGB1,anti-MyD88,and anti-TRIF could drastically attenuate the tissue injuries in ischemia reperfusion.anti-HMGB1 exhibited the most significant effect.Conclusions HMGB1 and its downstream signals play an important role in intestinal ischemia reperfusion injuries in mice.Of two downstream signals,the TRIF-dependent pathway exerts a more important effect than that of the MyD88-dependent pathway.

Protective effects of ferulic acid on mice bone marrow hematopoietic damage induced by irradiation / 中华放射医学与防护杂志

Objective To observe the effect and the mechanisms of ferulic acid on radiationinduced damage of mice peripheral blood and bone marrow hematopoietic function.Methods Ninety-six mice were randomly divided into sham irradiation group,irradiation group,positive drug group and 10,30,90 mg·kg-1 ·d-1 ferulic acid group,16 mice per group.Mice were exposed to 3.5 Gy γ-rays 24 h after first drug taken.Then,mice were given drugs for 7 d after irradiation.White blood cells in peripheral blood of 10 mice per group were counted 2 d before irradiation and 3,7,10,15 and 22 days after irradiation.The bone narrow of the other six mice was taken to detect the micronuclei frequency of polychromatic erythrocyte,the hematopoietic progenitor cell colony formation capacity,Thbd and HMGB1 protein expressions in mice bone marrow on the seventh day after irradiation.Results Compared with the irradiation alone group,the treatment of mice with ferulic acid 90 mg· kg-1 · d-1 increased the number of white blood cells in peripheral blood at 3,10,15 and 22 d after irradiation (t =2.267,2.399,1.945,2.828,P ＜ 0.05).Treatment with mice with ferulic acid 90 mg· kg-1 · d 1 decreased the micronuclei rate of erythrocytes in irradiated bone marrow (t =4.013,P ＜ 0.05),increased the clone numbers of CFU-E,BFU-E and CFU-GM of hematopoietic progenitor cells (t =2.366,2.953,3.115,P ＜0.05),improved the relative expression of the Thbd protein in bone marrow and the HMGB1 protein in nuclear (t =17.75,23.39,P ＜ 0.01).Conclusions Ferulilc acid could protect the bone marrow hematopoietic of mice exposed to irradiation by regulating the expressions of Thbd and HMGB1 protein,and then accelerate the peripheral cells recovery.

Updates on the Pathways of Toll-Like Receptors Mediated by HMGB1 / 现代检验医学杂志

High mobility group box 1 protein,a kind of damage-associated molecular patterns,is a group of nonhistone DNA-binding protein in the nuclear.It not only involves in DNA replication,recombination,repair,regulation of gene transcription,but also induces activation of innate immunity and T cell generation and activates the adaptive immune response through Toll-like receptor.It is also more closely related to autoimmune diseases,inflammatory diseases,tumors and so on.Currently there are many researches on the role of TLR-mediated HMGB1 signaling pathway in diseases,which is involved in a variety of pathological processes and has a broad clinical outlook.