Hmo1:A versatile member of the high mobility group box family of chromosomal architecture proteins

Eukaryotic chromatin consisting of nucleosomes connected by linker DNA is organized into higher order structures,which is facilitated by linker histone H1.Formation of chromatin compacts and protects the genome,but also hinders DNA transactions.Cells have evolved mechanisms to modify/remodel chromatin resulting in chromatin states suitable for genome functions.The high mobility group box(HMGB)proteins are non-histone chromatin architectural factors characterized by one or more HMGB motifs that bind DNA in a sequence nonspecific fashion.They play a major role in chromatin dynamics.The Saccharomyces cerevisiae(yeast hereafter)HMGB protein Hmo1 contains two HMGB motifs.However,unlike a canonical HMGB protein that has an acidic C-terminus,Hmo1 ends with a lysine rich,basic,C-terminus,resembling linker histone H1.Hmo1 exhibits characteristics of both HMGB proteins and linker histones in its multiple functions.For instance,Hmo1 promotes transcription by RNA polymerases I and II like canonical HMGB proteins but makes chromatin more compact/stable like linker histones.Recent studies have demonstrated that Hmo1 destabilizes/disrupts nucleosome similarly as other HMGB proteins in vitro and acts to maintain a common topological architecture of genes in yeast genome.This minireview reviews the functions of Hmo1 and the underlying mechanisms,highlighting recent discoveries.

High-mobility group box 1 protein and its role in severe acute pancreatitis

The high mobility group box 1(HMGB1),which belongs to the subfamily of HMG-1/-2,is a highly conserved single peptide chain consisting of 215 amino acid residues with a molecular weight of approximately 24894 Da.HMGB1 is a ubiquitous nuclear protein in mammals and plays a vital role in inflammatory diseases.Acute pancreatitis is one of the most common causes of acute abdominal pain with a poor prognosis.Acute pancreatitis is an acute inflammatory process of the pancreas(duration of less than six months),for which the severe form is called severe acute pancreatitis(SAP).More and more studies have shown that HMGB1 has a bidirectional effect in the pathogenesis of SAP.Extracellular HMGB1 can aggravate the pancreatic inflammatory process,whereas intracellular HMGB1 has a protective effect against pancreatitis.The mechanism of HMGB1 is multiple,mainly through the nuclear factor-κB pathway.Receptors for advanced glycation endproducts and toll-like receptors(TLR),especially TLR-2 and TLR-4,are two major types of receptors mediating the inflammatory process triggered by HMGB1 and may be also the main mediators in the pathogenesis of SAP.HMGB1 inhibitors,such as ethyl pyruvate,pyrrolidine dithiocarbamate and Scolopendra subspinipes mutilans,can decrease the level of extracellular HMGB1 and are the promising targets in the treatment of SAP.

High mobility group box-1 protein inhibits regulatory T cell immune activity in liver failure in patients with chronic hepatitis B

BACKGROUND: Liver failure in chronic hepatitis B (CHB) patients is a severe, life-threatening condition. Intestinal endotoxemia plays a significant role in the progress to liver failure. High mobility group box-1 (HMGB1) protein is involved in the process of endotoxemia. Regulatory T (Treg) cells maintain immune tolerance and contribute to the immunological hyporesponsiveness against HBV infection. However, the roles of HMGB1 and Treg cells in the pathogenesis of liver failure in CHB patients, and whether HMGB1 affects the immune activity of Treg cells are poorly known at present, and so were explored in this study. METHODS: The levels of HMGB1 expression were detected by ELISA, real-time RT-PCR, and Western blotting, and the percentage of CD4(+)CD25(+)CD127(low) Treg cells among CD4(+) cells was detected by flow cytometry in liver failure patients with chronic HBV infection, CHB patients, and healthy controls. Then, CD4(+)CD25(+)CD127(low) Treg cells isolated from the peripheral blood mononuclear cells from CHB patients were stimulated with HMGB1 at different concentrations or at various intervals. The effect of HMGB1 on the immune activity of Treg cells was assessed by a suppression assay of the allogeneic mixed lymphocyte response. The levels of forkhead box P3 (Foxp3) expression in Treg cells treated with HMGB1 were detected by RT-PCR and Western blotting. RESULTS: A higher level of HMGB1 expression and a lower percentage of Treg cells within the population of CIA(+) cells were found in liver failure patients than in CHB patients (82.6+/-20.1 mu g/L vs. 34.2+/-13.7 mu g/L; 4.55+/-1.34% vs. 9.52+/-3.89%, respectively). The immune activity of Treg cells was significantly weakened and the levels of Foxp3 expression were reduced in a dose- or time-dependent manner when Treg cells were stimulated with HMGB1 in vitro. CONCLUSIONS: The high level of HMGB1 and the low percentage of Treg cells play an important role in the pathogenesis of liver failure in patients with chronic HBV infection. Moreover, HMGB1 can weaken the immune activity of Treg cells. It is suggested that effectively inhibiting HMGB1 expression could be a feasible way to treat liver failure by suppressing endotoxemia and enhancing Treg cell activity.

Expression of high mobility group protein B1 in the lungs of rats with sepsis

BACKGROUND： Vibrio vulnifi cus inside the body could activate the NF-!B signaling pathwayand initiate the inflammatory cascade. The lung is one of the earliest organs affected by sepsisassociated with acute lung injury. High mobility group protein B1 （HMGB1） is an important late-actingpro-infl ammatory cytokine involving in the pathophysiology of sepsis. It is also involved in the injuryprocess in the lung, liver and intestine. There has been no report on the involvement of HMGB1 inVibrio vulnifi cus sepsis-induced lung injury.METHODS： Sixty rats were randomly divided into a normal control group （group A, n=10） anda Vibrio vulnificus sepsis group （group B, n=50）. Sepsis was induced in the rats by subcutaneousinjection of Vibrio vulnificus （concentration 6×108 cfu/mL, volume 0.1 mL/100g）） into the left lowerlimbs. The rats in group B were sacrifi ced separately 1, 6, 12, 24, and 48 hours after the infection.Their lungs were stored as specimens, lung water content was measured, and lung pathology wasobserved under a light microscope. The expressions of the HMGB1 gene and protein in the lungswere detected by RT-PCR and Western blot. Data were analyzed with one-way analysis of variance（ANOVA） and the LSD method for pair-wise comparison between the two groups. P〈0.05 wasconsidered statistically signifi cant.RESULTS： Compared to group A （0.652±0.177）, HMGB1 mRNA expression in the lungs ofgroup B was signifi cantly higher at 0 hour （1.161±0.358, P=0.013）, 24 hours （1.679±0.235, P=0.000）,and 48 hours （1.258±0.274, P=0.004） （P〈0.05）, and peaked at 24 hours. Compared to group A（0.594±0.190）, HMGB1 protein expression at 6 hours （1.408±0.567, P=0.026） after infection wassignificantly increased （P〈0. 05）, and peaked at 24 hours （2.415±1.064, P=0.000） after infection.Compared to group A （0.699±0.054）, lung water content was significantly increased at 6 hours（0.759±0.030, P=0.001）,12 hours （0.767±0.023, P=0.000）, 24 hours （0.771±0.043, P=0.000） and 48hours （0.789±0.137, P=0.000） after infection （P〈0.05）. Compared to group A, pathological changesat 12 hours in group B indicate marked pulmonary vascular congestion, interstitial edema andinfl ammatory infi ltration. Alveolar cavity collapse and boundaries of the alveolar septum could not beclearly identifi ed.CONCLUSION： Vibrio vulnifi cus sepsis can lead to injury in rat lungs, and increased HMGB1expression in lung tissue may be one of the mechanisms for injury from Vibrio vulnifi cus sepsis.

Novel insights for high mobility group box 1 proteinmediated cellular immune response in sepsis:A systemic review

High mobility group box 1 protein （HMGB1） is a highly conserved, ubiquitous protein in the nuclei and cytoplasm of nearly all cell types. HMGB1 is secreted into the extracellular milieu and acts as a proinflammatory cytokine. In this article we reviewed briefly the cellular immune response mediated by HMGB1 in inflammation and sepsis. This systemic review is mainly based on our own work and other related reports HMGB1 can actively affect the immune functions of many types of cells including T lymphocytes, regulatory T cells （Tregs）, dendritic cells （DCs）, macrophages, and natural killer cells （NK cells）. Various cellular responses can be mediated by HMGB1 which binds to cell-surface receptors [e.g., the receptor for advanced glycation end products （RAGE）, Toll-like receptor （TLR）2, and TLR4]. Anti-HMGB1 treatment, such as anti-HMGB1 polyclonal or monoclonal antibodies, inhibitors （e.g., ethyl pyruvate） and antagonists （e.g., A box）, can protect against sepsis lethality and give a wider window for the treatment opportunity. HMGB1 is an attractive target for the development of new therapeutic strategies in the treatment of patients with septic complications.

Inflammatory response and immune regulation of high mobility group box-1 protein in treatment of sepsis

Sepsis is an infection induced systemic inflammatory response syndrome and is a major cause of morbidity as well as mortality in intensive care units. A growing body of evidence suggests that the activation of a proinflammatory cascade is responsible for the development of immune dysfunction, susceptibility to severe sepsis and septic shock. The present theories of sepsis as a dysregulated inflammatory response and immune function, as manifested by excessive release of inflammatory mediators such as high mobility group box 1 protein （HMGB1）, are supported by increasing studies employing animal models and clinical observations of sepsis. HMGB1, originally described as a DNA-binding protein and released passively by necrotic cells and actively by macrophages/monocytes, has been discovered to be one of essential cytokines that mediates the response to infection, injury and inflammation. A growing number of studies still focus on the inflammation-regulatory function and its contribution to infectious and inflammatory disorders, recent data suggest that HMGB1 formation can also markedly influence the host cell-mediated immunity, including T lymphocytes and macrophages. Here we review emerging evidence that support extracellular HMGB1 as a late mediator of septic complications, and discuss the therapeutic potential of several HMGBl-targeting agents in experimental sepsis. In addition, with the development of traditional Chinese medicine in recent years, it has been proven that traditional Chinese herbal materials and their extracts have remarkable effective in treating severe sepsis. In this review, we therefore provide some new concepts of HMGBl-targeted Chinese herbal therapies in sepsis.

Scolopendra subspinipes mutilans protected the ceruleininduced acute pancreatitis by inhibiting high-mobility group box protein-1

AIM:To evaluate the inhibitory effects of Scolopendra subspinipes mutilans(SSM) on cerulein-induced acute pancreatitis(AP) in a mouse model.METHODS:SSM water extract(0.1,0.5,or 1 g/kg) was administrated intraperitoneally 1 h prior to the first injection of cerulein.Once AP developed,the stable cholecystokinin analogue,cerulein was injected hourly,over a 6 h period.Blood samples were taken 6 h later to determine serum amylase,lipase,and cytokine levels.The pancreas and lungs were rapidly removed for morphological examination,myeloperoxidase assay,and real-time reverse transcription polymerase chain reaction.To specify the role of SSM in pancreatitis,the pancreatic acinar cells were isolated using collagenase method.Then the cells were pre-treated with SSM,then stimulated with cerulein.The cell viability,cytokine productions and high-mobility group box protein-1(HMGB-1) were measured.Furthermore,the regulating mechanisms of SSM action were evaluated.RESULTS:The administration of SSM significantly attenuated the severity of pancreatitis and pancreatitis associated lung injury,as was shown by the reduction in pancreatic edema,neutrophil infiltration,vacuolization and necrosis.SSM treatment also reduced pancreatic weight/body weight ratio,serum amylase,lipase and cytokine levels,and mRNA expression of multiple inflammatory mediators such as tumor necrosis factor-α and interleukin-1β.In addition,treatment with SSM inhibited HMGB-1 expression in the pancreas during AP.In accordance with in vivo data,SSM inhibited the cerulein-induced acinar cell death,cytokine,and HMGB-1 release.SSM also inhibited the activation of c-Jun NH2-terminal kinase,p38 and nuclear factor(NF)-κB.CONCLUSION:These results suggest that SSM plays a protective role during the development of AP and pancreatitis associated lung injury via deactivating c-Jun NH2-terminal kinase,p38 and NF-κB.

Suppressing high mobility group box-1 release alleviates morphine tolerance via the adenosine5'-monophosphate-activated protein kinase/heme oxygenase-1 pathway

Opioids,such as morphine,are the most potent drugs used to treat pain.Long-term use results in high tolerance to morphine.High mobility group box-1(HMGB1) has been shown to participate in neuropathic or inflammatory pain,but its role in morphine tolerance is unclear.In this study,we established rat and mouse models of morphine tolerance by intrathecal injection of morphine for 7 consecutive days.We found that morphine induced rat spinal cord neurons to release a large amount of HMGB1.HMGB1 regulated nuclear factor κB p65 phosphorylation and interleukin-1β production by increasing Toll-like receptor 4receptor expression in microglia,thereby inducing morphine tolerance.Glycyrrhizin,an HMGB1 inhibito r,markedly attenuated chronic morphine tole rance in the mouse model.Finally,compound C(adenosine 5’-monophosphate-activated protein kinase inhibitor) and zinc protoporphyrin(heme oxygenase-1 inhibitor)alleviated the morphine-induced release of HMGB1 and reduced nuclear factor κB p65 phosphorylation and interleukin-1β production in a mouse model of morphine tolerance and an SH-SY5Y cell model of morphine tole rance,and alleviated morphine tolerance in the mouse model.These findings suggest that morphine induces HMGB1 release via the adenosine 5’-monophosphate-activated protein kinase/heme oxygenase-1 signaling pathway,and that inhibiting this signaling pathway can effectively reduce morphine tole rance.

Effect of high mobility group box-1 protein on immune cells and its regulatory mechanism

High mobility group box-1 protein(HMGB1),which is a nuclear protein,participates in chromatin architecture and transcriptional regulation.When released from cells,HMGB1 also plays a well-established role as a pro-inflammatory mediator during innate immune responses to injury.In the initial stage of injury,there is a release of large quantities of early pro-inflammatory mediators to initiate or perpetuate immune responses against pathogens,but this pro-inflammatory period is transient,and it is followed by a prolonged period of immune suppression.At present,several lines of evidences have suggested that HMGB1 is a late cytokine provoking delayed endotoxin morbidity,which may enhance the production of early proinflammatory mediators,and it can contribute potently to the activation of different immune cells and play a role in the development of host cell-mediated immunity.The biology of HMGB1 has been extensively studied as a pro-inflammatory cytokine of systemic inflammation,however,this review will attempt to provide a summary of the effects of HMGB1 on different immune cells and its regulatory mechanism in acute insults.

Relationship of plasma homocysteine, soluble intercellular adhesion molecule-1, high mobility group box 1 protein with carotid intima-media thickness in elderly patients with type 2 diabetes mellitus

Objective:To explore the relationship of plasma homocysteine(Hcy),soluble intercellular adhesion molecule-1(sICAM-1)and high mobility group box 1 protein(HMGB1)with carotid intima-media thickness(c-IMT)in elderly patients with type 2 diabetes mellitus.Methods:A total of 100 elderly patients who were diagnosed as type 2 diabetes mellitus in Baogang Hospital of Inner Mongolia from June 2017 to May 2020 were chosen as research objects.According to c-IMT,they were divided into the normal group(n=35),the mild to moderate group(n=41)and the severe group(n=24).The expression levels of plasma Hcy,sICAM-1 and HMGB1 were compared between groups respectively.Pearson’s correlation coefficient was used to analyze the relationship of plasma Hcy,sICAM-1,HMGB1 with c-IMT.Results:The comparison in plasma Hcy,sICAM-1,HMGB1 and c-IMT among the three groups of patients was of statistical significance(p<.05).The results of correlation analysis showed that the expression levels of plasma Hcy,sICAM-1 and HMGB1 were positively correlated with c-IMT in elderly patients with type 2 diabetes mellitus(r=.627,.598,.614;p<.05).Conclusions:The expression levels of plasma Hcy,sICAM-1 and HMGB1 are abnormally increased in elderly patients with type 2 diabetes mellitus,and related to c-IMT,which can provide a strong evidence for clinical diagnosis and treatment by detecting their levels in clinical practice.

Renal Expression and Clinical Significance of High Mobility Group Box 1 in Lupus Nephritis

Objective: To evaluate the clinical significance of high mobility group box 1 (HMGB1) expression in nephridial tissues of lupus nephritis (LN). Methods: Sixty-three patients with active LN and 15 systemic lupus erythematosus (SLE) (combined without LN) were included. Renal biopsies were performed in the two groups. The biopsies were evaluated according to the World Health Organization (WHO) classification and renal disease activity was estimated using the British Isles lupus assessment group (BILAG) index. Serum levels of HMGB1 were analyzed by western blot. HMGB1 expression in renal tissue was assessed by immunohistochemistry in the two groups. The correlation between HMGB1 and renal active index (AI), chronicity index (CI), pathological type of LN was analyzed. Results: LN biopsies showed WHO class III, IV or V and all patients had high renal disease activity (BILAG A/B). The HMGB1 expression was higher in the LN groups than the control groups (t = 9.263, P < 0.05). It showed positive correlation between HMGB1 expression and SLE DAI classification (r = 0.579, P P < 0.05) and renal tubule interstitial (TIL) classification (r = 0.815, P < 0.05), and negative correlation between HMGB1 expression and CI classification (r = 0.582, P < 0.05). In all patients, serum levels of HMGB1 increased only slightly in the patients only with SLE;however, in patients with LN WHO class IV a significant decrease was observed (P = 0.02). Immunostaining revealed a pronounced extranuclear HMGB1 expression predominantly outlining the glomerular endothelium and in the mesangium. There were significant differences in HMGB1 expression between LN and control biopsies and it existed with apparent association to histopathological classification and clinical outcome. Conclusions: Renal tissue expression and serum levels of HMGB1 were elevated in LN. The unusual elevation of HMGB1 in serum and tissue in LN may reflect persistent inflammatory activity, which clearly indicates a role for HMGB1 in pathogenesis of LN.

Expression of HMGB1 Protein in Human Cervical Squamous Epithelium Carcinoma

OBJECTIVE To investigate the expression of the high mobility group boxl（HMGB1） in human cervical squamous epithelial carcinoma （CSEC） and to explore the relationship of HMGB1 expression to the differentiation degree, size, invasion and metastasis of CSEC. METHODS Immunohistochemical staining of tissue microarrays and Western blot analysis were conducted to detect the expression of HMGB1 in the following tissue samples： 30 carcinoma in situ, 90 invasive CSEC without metastasis, 30 invasive CSEC with metastasis, 30 cases of normal cervical squamous epithelia. RESULTS The positive-expression rate of HMGB1 was 58.7% （88/150） in CSEC, showing a significant difference compared to normal cervical squamous epithelia. The expression of HMGB1 was correlated with tumor size, invasion and metastasis of CSEC （respectively, P〈0.01）, but had no relationship with the degree of differentiation （P〉0.05）. CONCLUSION The over-expression of HMGB1 in CSEC might be a useful parameter as an indication of tumor invasion, metastasis, prognosis and overall biological behavior of human CSEC, as well as a noval target site for gene therapy.

血清Ang-2和HMGB1水平与脓毒症患者病情及预后相关性的研究

目的 探究血清血管生成素-2(Ang-2)和高迁移率族蛋白B1(HMGB1)水平与脓毒症患者病情及28天预后的相关性。方法 回顾性分析41例脓毒症患者资料,根据患者28天的预后情况分为存活组和死亡组;根据患者病情严重程度将2组患者分别分为脓毒症组和脓毒性休克组。记录所有入组患者的一般资料并进行第1天、第3天、第5天及第7天的APACHEⅡ评分,测定Ang-2和HMGB1水平。结果 存活组与死亡组患者的一般资料比较差异无统计学意义。死亡组脓毒症休克患者占比及入科APACHEⅡ评分为(85.71%)和(29.29±5.99)分,均明显高于存活组(51.84%)和(24.96±5.84)分。2组患者血清Ang-2和HMGB1水平比较差异无统计学意义。第7天的HMGB1水平与脓毒症患者预后显著相关,而第1天、第3天、第5天的HMGB1水平及上述各时间点的Ang-2水平与患者预后均无相关性。各时间点血清Ang-2水平的算术平均值与APACHEⅡ评分的算术平均值呈正相关,HMGB1水平与APACHEⅡ评分并无相关性。联合第7天的APACHEⅡ评分、Ang-2和HMGB1水平的曲线下面积最大为0.78,灵敏度为92.9%,特异度为59.3%。联合第7天的Ang-2和HMGB1水平预测预后不良的曲线下面积为0.71,灵敏度为92.9%,特异度为55.6%。结论 脓毒症患者第1天、第3天、第5天及第7天的平均血清Ang-2水平与病情严重程度呈正相关;联合脓毒症患者第7天的血清Ang-2和HMGB1水平对其预后进行评估灵敏度最高,明显高于单独某项指标的评估价值。

血清SP-A、SP-D、HMGB1、KL-6、miR-21与老年急性加重特发性肺纤维化及肺功能的关系

目的 探讨血清表面活性蛋白-A(SP-A)、表面活性蛋白-D(SP-D)、高迁移率族蛋白B1(HMGB1)、涎液化糖链抗原-6(KL-6)、微小核糖核酸-21(miR-21)与老年急性加重特发性肺纤维化(AE-IPF)及肺功能的关系,以提高AE-IPF早期诊断率、防范误诊的发生。方法 选取2023年1月至2024年1月收治的老年AE-IPF患者80例为AE-IPF组,另选择同期就诊的80例老年稳定特发性肺纤维化(IPF)患者为IPF组及50例体检健康老年人为对照组。比较3组血清SP-A、SP-D、HMGB1、KL-6、miR-21及肺功能指标[第1秒用力呼气容积(FEV1)、FEV1/用力肺活量(FVC)、第1秒用力呼气容积占预计值百分比(FEV1%pred)、1 min最大自主通气量(MVV)、1 min最大自主通气量占预计值百分比(MVV%pred)、肺一氧化碳弥散量占预计值百分比(DLCO%pred)]。分析血清SP-A、SP-D、HMGB1、KL-6、miR-21诊断AE-IPF的价值;分析血清SP-A、SP-D、HMGB1、KL-6、miR-21与肺功能指标的相关性。结果 IPF组、AE-IPF组血清SP-A、SP-D、HMGB1、KL-6、miR-21高于对照组,且AE-IPF组高于IPF组(P<0.05)。血清SP-A、SP-D、HMGB1、KL-6、miR-21诊断AE-IPF发生的曲线下面积分别为0.866、0.882、0.890、0.793、0.783,敏感度分别为0.800、0.825、0.800、0.763、0.813。AE-IPF组、IPF组FEV1、FEV1/FVC、FEV1%pred、MVV、MVV%pred、DLCO%pred均低于对照组,AE-IPF组低于IPF组(P<0.05)。血清SP-A、SP-D、HMGB1、KL-6、miR-21与FEV1、FEV1/FVC、FEV1%pred、MVV、MVV%pred、DLCO%pred均呈负相关(P<0.01)。结论 检测血清SP-A、SP-D、HMGB1、KL-6、miR-21表达情况可提高AE-IPF早期诊断率,防范误诊误治的发生。

儿童肺炎支原体肺炎肺泡灌洗液CARDS TX、HMGB1、TLR2表达及临床意义

目的研究肺炎支原体肺炎(MPP)患儿肺泡灌洗液中社区获得性呼吸窘迫综合征毒素(CARDS TX)、高迁移率族蛋白1(HMGB1)、Toll样受体2(TLR2)表达,探讨其在MPP发病中的临床意义。方法回顾性分析55例MPP病例组及20例对照组临床资料,同时检测两组支气管肺泡灌洗液(BALF)中CARDS TX、HMGB1、TLR2、TLR4、晚期糖基化终末产物受体(RAGE)及髓样分化因子88(MyD88)表达,体外检测不同浓度CARDS TX刺激下HMGB1、TLR2的表达并进行比较。结果与对照组相比,MPP组LYM绝对值计数、PA、CD3^(+)、CD3^(+)CD4^(+)、CD3^(+)CD8^(+)、CD3-CD19^(+)、NK cell、CD19^(+)CD23^(+)明显下降,CRP、LDH、D-二聚体、IgA、IgG、IgM、CARDS TX、HMGB1、TLR2、MyD88均升高,差异有统计学意义(P<0.05或0.001)。CARDS TX刺激后HMGB1、TLR2的表达升高,且呈正相关,差异有统计学意义(P<0.001)。结论CARDS TX可能通过HMGB1-TLR2-MyD88途径参与肺炎支原体(MP)的致病。

穿心莲内酯调节HMGB1/RAGE信号通路对糖尿病周围神经病变大鼠坐骨神经功能损伤的影响

目的探讨穿心莲内酯调节高迁移率族蛋白B1(HMGB1)/晚期糖基化终产物受体(RAGE)信号通路对糖尿病周围神经病变(DPN)大鼠坐骨神经功能损伤的影响。方法将84只大鼠随机分为对照组(生理盐水)、DPN组(生理盐水)、穿心莲内酯低剂量组(0.833 mg/kg)、穿心莲内酯高剂量组(3.332 mg/kg)、硫辛酸组(阳性对照,0.1 g/kg)、重组大鼠HMGB1蛋白(rHMGB1,8μg/kg)组、穿心莲内酯高剂量+rHMGB1组,每组12只。除对照组外,其余各组大鼠均采用高糖高脂饲料喂养联合腹腔注射链脲佐菌素的方式构建DPN模型。造模成功24 h后,进行给药处理,每天1次,持续8周。给药结束后,检测大鼠空腹血糖、机械痛阈值、热痛阈值、坐骨神经传导速度的变化;观察大鼠坐骨神经病理变化;检测大鼠坐骨神经中超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量;检测大鼠坐骨神经中HMGB1、RAGE蛋白表达水平和核因子κB p65(NF-κB p65)蛋白磷酸化水平。结果与对照组比较,DPN组大鼠坐骨神经病理损伤严重,空腹血糖、热痛阈值、MDA含量及HMGB1、RAGE蛋白表达水平和NF-κB p65蛋白磷酸化水平均显著升高(P<0.05),机械痛阈值、感觉神经传导速度、运动神经传导速度、SOD活性显著降低/减慢(P<0.05);与DPN组比较,穿心莲内酯低、高剂量组和硫辛酸组大鼠上述指标均显著改善(P<0.05),rHMGB1组对应指标变化趋势与上述3个给药组相反(P<0.05);并且,rHMGB1可减弱高剂量穿心莲内酯对DPN大鼠血糖的降低作用及坐骨神经氧化应激损伤的改善作用(P<0.05)。结论穿心莲内酯可能通过抑制HMGB1/RAGE信号通路来降低血糖、抑制氧化应激,进而改善DPN大鼠坐骨神经损伤。

术前血清sTim-3、HMGB1水平与肌层浸润性膀胱癌根治术后预后的关系

目的探讨术前血清可溶性T细胞免疫球蛋白黏蛋白分子-3(sTim-3)、高迁移率族蛋白B1(HMGB1)水平与肌层浸润性膀胱癌(MIBC)根治术后预后的关系。方法选取2019年6月至2020年6月该院收治的85例MIBC患者作为MIBC组,另选取同期在该院体检的85例健康者作为对照组。采用酶联免疫吸附试验(ELISA)检测所有受试者血清sTim-3、HMGB1水平;根据血清sTim-3、HMGB1水平均值将MIBC患者分为sTim-3高表达组(≥均值)和sTim-3低表达组(<均值)、HMGB1高表达组(≥均值)和HMGB1低表达组(<均值)。对比各组血清sTim-3、HMGB1水平,以及不同sTim-3、HMGB1水平与MIBC患者病理特征的关系。采用Pearson相关分析MIBC患者血清sTim-3水平与血清HMGB1水平的相关性;采用Kaplan-Meier生存曲线分析不同血清sTim-3、HMGB1水平MIBC患者的生存预后情况。结果MIBC组患者血清sTim-3、HMGB1水平高于对照组(P<0.05)。Pearson相关性分析结果显示,MIBC组患者血清sTim-3水平与血清HMGB1水平呈正相关(r=0.405,P<0.001)。不同年龄、性别、肿瘤最大径及是否发生淋巴结转移MIBC患者的血清sTim-3、HMGB1水平比较,差异均无统计学意义(P>0.05),而不同TNM分期、组织分级、淋巴结状态MIBC患者的血清sTim-3、HMGB1水平比较,差异均有统计学意义(P<0.05)。根据血清sTim-3、HMGB1水平的均值将85例MIBC患者分为sTim-3高表达组(≥3.67 ng/mL,n=44)和sTim-3低表达组(<3.67 ng/mL,n=41)、HMGB1高表达组(≥14.91 ng/mL,n=47)和HMGB1低表达组(<14.91 ng/mL,n=38)。Kaplan-Meier生存曲线结果显示,sTim-3低表达组患者的3年生存曲线高于sTim-3高表达组患者(Log-rankχ^(2)=6.175,P=0.013),HMGB1低表达组患者的3年生存曲线高于HMGB1高表达组患者(Log-rankχ^(2)=4.056,P=0.044)。sTim-3高表达组与sTim-3低表达组MIBC患者的3年生存率分别为52.27%、78.05%,HMGB1高表达组与HMGB1低表达组MIBC患者3年生存率分别为55.32%、76.31%。结论血清sTim-3、HMGB1在MIBC患者中呈高表达,且二者水平呈正相关,可作为评估MIBC患者预后不良的重要指标。

MiR-142-3p Regulates ILC1s by Targeting HMGB1 via the NF-κB Pathway in a Mouse Model of Early Pregnancy Loss

Objective Innate lymphoid cells(ILCs)are a class of newly discovered immunocytes.Group 1 ILCs(ILC1s)are identified in the decidua of humans and mice.High mobility group box 1(HMGB1)is predicted to be one of the target genes of miR-142-3p,which is closely related to pregnancy-related diseases.Furthermore,miR-142-3p and HMGB1 are involved in regulating the NF-κB signaling pathway.This study aimed to examine the regulatory effect of miR-142-3p on ILC1s and the underlying mechanism involving HMGB1 and the NF-κB signaling pathway.Methods Mouse models of normal pregnancy and abortion were constructed,and the alterations of ILC1s,miR-142-3p,ILC1 transcription factor(T-bet),and pro-inflammatory cytokines of ILC1s(TNF-α,IFN-γand IL-2)were detected in mice from different groups.The targeting regulation of HMGB1 by miR-142-3p in ILC1s,and the expression of HMGB1 in normal pregnant mice and abortive mice were investigated.In addition,the regulatory effects of miR-142-3p and HMGB1 on ILC1s were detected in vitro by CCK-8,Annexin-V/PI,ELISA,and RT-PCR,respectively.Furthermore,changes of the NF-κB signaling pathway in ILC1s were examined in the different groups.For the in vivo studies,miR-142-3p-Agomir was injected in the uterus of abortive mice to evaluate the abortion rate and alterations of ILC1s at the maternal-fetal interface,and further detect the expression of HMGB1,pro-inflammatory cytokines,and the NF-κB signaling pathway.Results The number of ILC1s was significantly increased,the level of HMGB1 was significantly upregulated,and that of miR-142-3p was considerably downregulated in the abortive mice as compared with the normal pregnant mice(all P<0.05).In addition,miR-142-3p was found to drastically inhibit the activation of the NF-κB signaling pathway(P<0.05).The number of ILC1s and the levels of pro-inflammatory cytokines were significantly downregulated and the activation of the NF-κB signaling pathway was inhibited in the miR-142-3p Agomir group(all P<0.05).Conclusion miR-142-3p can regulate ILC1s by targeting HMGB1 via the NF-κB signaling pathway,and attenuate the inflammation at the maternal-fetal interface in abortive mice.

HGI、β2-MG、HMGB1在2型糖尿病视网膜病变早期诊断中的临床价值

目的 探讨糖化血红蛋白变异指数(HGI)、血清β2微球蛋白(β2-MG)及高迁移族蛋白B1(HMGB1)在2型糖尿病视网膜病变(DR)早期诊断中的临床价值。方法 回顾性选取2020年1月~2023年1月在皖西卫生职业学院附属阜南县中医院治疗的2型糖尿病(T2DM)合并DR患者112例作为观察组,同时选取同期在本院治疗的单纯T2DM患者54例作为对照组,比较两组患者一般资料及HGI、β2-MG、HMGB1水平,多元logistic回归模型分析影响T2DM患者并发DR的危险因素,绘制受试者工作特征(ROC)曲线,分析HGI、β2-MG、HMGB1三者单独及联合检测对T2DM患者并发DR的诊断效能。结果 观察组合并高血压占比率高于对照组,差异有统计学意义(χ^(2)=5.372,P<0.05);观察组T2DM病程、FPG、HbA1c、LDL-C、HGI、β2-MG、HMGB1水平均高于对照组,差异均有统计学意义(t=5.303、2.711、3.135、2.391、14.958、11.028、5.695,P<0.05)。多元logistic回归分析显示,合并高血压、HbA1c、HGI、β2-MG、HMGB1为影响T2DM患者并发DR的危险因素(P<0.05);ROC曲线结果显示,HGI、β2-MG、HMGB1单独T2DM患者并发DR的曲线下面积(AUC)分别为0.822、0.785、0.753,而三者联合检测的AUC为0.909,高于单一检测(P<0.05)。结论 相较于单纯T2DM患者,合并DR的T2DM患者的HGI、β2-MG、HMGB1水平异常升高,且上述指标均为DR发生的危险因素,通过联合检测三者水平对DR具有较好的诊断效能。

Overexpression of HMGB1 A-box reduced lipopolysaccharide-induced intestinal inflammation via HMGB1/TLR4 signaling in vitro

AIM: To investigate the inhibitory effects and mechanism of high mobility group box(HMGB)1 A-box in lipopolysaccharide(LPS)-induced intestinal inflammation.METHODS: Overexpression of HMGB1 A-box in human intestinal epithelial cell lines(SW480 cells) was achieved using the plasmid p EGFP-N1. HMGB1 A-box-overexpressing SW480 cells were stimulated with LPS and co-culturing with human monocyte-like cell lines(THP-1 cells) using a Transwell system, compared with another HMGB1 inhibitor ethyl pyruvate(EP). The m RNA and protein levels of HMGB1/toll-like receptor(TLR) 4 signaling pathways [including HMGB1, TLR4, myeloid differentiation factor88(MYD88), Phosphorylated Nuclear Factor κB(p NF-κB) p65] in the stimulated cells were determined by realtime polymerase chain reaction and Western blotting. The levels of the proinflammatory mediators [including HMGB1, interleukin(IL)-1β, IL-6 and tumor necrosis factor(TNF)-α] in the supernatants of the stimulated cells were determined by ELISA.RESULTS: EP downregulated the m RNA and protein levels of HMGB1, inhibited the TLR4 signaling pathways(TLR4, MYD88 and p NF-κB p65) and reduced the secretion of proinflammatory mediators(HMGB1, IL-1β, IL-6 and TNF-α) in the SW480 and THP-1 cells activated by LPS but not in the unstimulated cells. Activated by LPS, the overexpression of HMGB1 A-box in the SW480 cells also inhibited the HMGB1/TLR4 signaling pathways and reduced the secretion of these proinflammatory mediators in the THP-1 cells but not in the transfected and unstimulated cells. CONCLUSION: HMGB1 A-box, not only EP, can reduce LPS-induced intestinal inflammation through inhibition of the HMGB1/TLR4 signaling pathways.