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.

一类随机Box-样条的逼近问题

箱样条具有良好的性质和特殊的结构,是进行各种重构工作的有力工具.而客观世界里,有很多数据是随机地给出的,这就需要对其在随机层面进行研究.本文构造了一类特殊的随机Box-样条,并讨论了其对随机函数的逼近问题.

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.

High mobility group box-1 release from H2O2-injured hepatocytes due to sirt1 functional inhibition

BACKGROUND High mobility group box-1 (HMGB1), recognized as a representative of damageassociated molecular patterns, is released during cell injury/death, triggering the inflammatory response and ultimately resulting in tissue damage. Dozens of studies have shown that HMGB1 is involved in certain diseases, but the details on how injured hepatocytes release HMGB1 need to be elicited. AIM To reveal HMGB1 release mechanism in hepatocytes undergoing oxidative stress. METHODS C57BL6/J male mice were fed a high-fat diet for 12 wk plus a single binge of ethanol to induce severe steatohepatitis. Hepatocytes treated with H2O2 were used to establish an in vitro model. Serum alanine aminotransferase, liver H2O2 content and catalase activity, lactate dehydrogenase and 8-hydroxy-2- deoxyguanosine content, nicotinamide adenine dinucleotide (NAD+) levels, and Sirtuin 1 (Sirt1) activity were detected by spectrophotometry. HMGB1 release was measured by enzyme linked immunosorbent assay. HMGB1 translocation was observed by immunohistochemistry/immunofluorescence or Western blot. Relative mRNA levels were assayed by qPCR and protein expression was detected by Western blot. Acetylated HMGB1 and poly(ADP-ribose)polymerase 1 (Parp1) were analyzed by Immunoprecipitation. RESULTS When hepatocytes were damaged, HMGB1 translocated from the nucleus to the cytoplasm because of its hyperacetylation and was passively released outside both in vivo and in vitro. After treatment with Sirt1-siRNA or Sirt1 inhibitor (EX527), the hyperacetylated HMGB1 in hepatocytes increased, and Sirt1 activity inhibited by H2O2 could be reversed by Parp1 inhibitor (DIQ). Parp1 and Sirt1 are two NAD+-dependent enzymes which play major roles in the decision of a cell to live or die in the context of stress . We showed that NAD+ depletion attributed to Parp1 activation after DNA damage was caused by oxidative stress in hepatocytes and resulted in Sirt1 activity inhibition. On the contrary, Sirt1 suppressed Parp1 by negatively regulating its gene expression and deacetylation. CONCLUSION The functional inhibition between Parp1 and Sirt1 leads to HMGB1 hyperacetylation, which leads to its translocation from the nucleus to the cytoplasm and finally outside the cell.

DEAD box-1在儿童常见肿瘤的研究进展

DEAD box-1是DEAD-box RNA解旋酶家族成员之一,在细胞内该酶能水解NTP,与其他蛋白组成复合体发挥作用。参与核糖体、RNA 或DNA的结构重塑,影响RNA的生成及RNA的多态性。目前,越来越多的研究表明,DEAD-box 1在肿瘤的发生、发展中起着重要作用。MYCN基因扩增与多种儿童肿瘤密切相关,是儿童常见肿瘤的重要标记物,研究表明,DEAD box-1基因和MYCN基因在肿瘤中存在共扩增现象。本文根据DEAD box-1基因与儿童常见肿瘤的研究进展,对DEAD box-1与肿瘤关系进行综述。

去整合素金属蛋白酶10和高迁移率族蛋白B1在声门型喉癌患者中的表达及预后分析

目的探讨去整合素金属蛋白酶10(a disintegrin and metalloprotease 10,ADAM10)和高迁移率族蛋白B1(high mobility group box-1 protein,HMGB1)与声门型喉癌患者病理特征及预后关系分析。方法回顾性收集2017年3月~2020年12月于南京医科大学附属南京医院确诊及治疗的声门型喉癌患者50例(观察组),另取相对喉癌组织切缘0.5cm以上部位标本作为对照组。观察并比较ADAM10和HMGB1在两组中的阳性表达率,分析其阳性表达与声门型喉癌患者的病理特征关系。单因素分析影响声门型喉癌预后的危险因素,Cox多因素回归分析声门型喉癌患者不良预后的独立危险因素。结果ADAM10和HMGB1在观察组的阳性表达率均高于对照组,差异均有统计学意义(P<0.05)。声门型喉癌组织中的ADAM10与淋巴结转移和T分级差异比较有统计学意义,而与年龄、性别、饮酒史、吸烟史、分化程度差异比较无统计学意义(P>0.05);HMGB1与分化程度差异比较有统计学意义(P<0.05),而与年龄、性别、饮酒史、吸烟史、淋巴结转移、T分级差异比较无统计学意义(P>0.05)。单因素分析结果表明,淋巴结转移、T分级、分化程度、ADAM10、HMGB1是患者预后的影响因素。Cox多因素回归分析结果表明,淋巴结转移、T3+T4分级、低分化程度、ADAM10阳性、HMGB1阳性为声门型喉癌患者预后不良的独立影响因素(P<0.05)。结论ADAM10和HMGB1可作为声门型喉癌不良预后的风险评估指标。

Predictive Value of High Mobility Group Box-1 and miR-146b in Septic Shock Patients

In the face of the elevated incidence and mortality rate of septic shock in the ICU,this retrospective study seeks to investigate the indicative and predictive value of high-mobility group box 1(HMGB1)and miR-146b in patients with septic shock.Quantitative RTPCR was employed in this study to quantify the HMGB1 and miR-146b levels in plasma samples obtained from the patient group and healthy controls.The investigation involved the comparison between the two groups and tracking changes in the patient group over time.The finding revealed that upon admission,the patient group exhibited markedly elevated relative expression levels of HMGB1,which subsequently decreased over time.Conversely,the patient group displayed significantly reduced relative expression levels of miR-146b upon admission,which subsequently increased over time compared to the control group.Receiver operating characteristic(ROC)curves showed good predictive value for HMGB1 and miR-146b.The experimental results suggest that HMGB1 and miR-146b serve as valuable and convenient biomarkers for evaluating the severity of septic shock and predicting mortality.Additionally,it is proposed that serum miR-146b may be inducible and potentially exerts a negative regulatory effect on the expression of HMGB1.

血清降钙素原、HMGB1检测在脓毒症患儿中的临床意义

目的:分析血清降钙素原、高迁移率蛋白box-1(High mobility protein box-1,HMGB1)检测在脓毒症患儿中的临床意义。方法:选取2020年4月至2022年5月本院收治的105例脓毒症患儿作为研究对象的脓毒症组,根据病情严重程度分为脓毒症组(n=46)、严重脓毒症组(n=28)和感染性休克组(n=31);另选取同期本院收治的43例一般细菌性感染患儿作为研究对象的非脓毒症组。比较不同人群、不同病情程度血清降钙素原、HMGB1水平及急性生理功能和慢性健康状况评分系统Ⅱ评分(Acute physiological function and Chronic health status score System II score,APACHEⅡ),采用Pearson相关分析血清降钙素原、HMGB1水平与APACHEⅡ评分相关性。患儿出院后进行3 m随访观察脓毒症患儿预后情况,且测定不同预后血清降钙素原、HMGB1水平。结果:脓毒症组、严重脓毒症组及感染性休克组的降钙素原、HMGB1水平及APACHEⅡ评分明显高于非脓毒症组;脓毒症病情越严重,血清降钙素原、HMGB1水平及APACHEⅡ评分越高(P<0.05);Pearson相关分析结果显示,血清降钙素原、HMGB1水平与APACHEⅡ评分呈正相关(P<0.05)。随访结果显示,患儿预后良好99例、预后不良6例;治疗3 m后,预后不良组血清降钙素原、HMGB1水平明显高于预后良好组(P<0.05)。结论:在临床中及时检测血清降钙素原、HMGB1,可尽早评判脓毒症患儿病情发展情况,且有效预测预后。

肾茶总黄酮的提取纯化工艺优化

目的应用Box-Behnken效应面法优化肾茶提取工艺,运用大孔吸附树脂优选肾茶总黄酮的纯化工艺。方法以肾茶总黄酮得率为评价指标,以乙醇体积分数,液料比和提取时间为考察因素,采用Box-Behnken效应面法优化加热回流提取工艺,并进行预测分析。通过静态和动态吸附试验考察大孔吸附树脂对肾茶总黄酮的纯化效果,并研究各因素条件对其纯化效果的影响,筛选主要影响指标的最佳工艺条件。结果肾茶最佳提取工艺为:乙醇体积分数为52.52%,液料比(mL:g)为15:1,提取时间为90分钟。DM-130大孔吸附树脂较适合肾茶总黄酮的纯化,静态吸附生药浓度为0.1 g/mL,药液pH值为2;动态吸附上样量为60 mL,吸附时间为50分钟,除杂水用量为3倍柱体积,洗脱剂为70%乙醇,洗脱剂用量为3倍树脂体积。结论 Box-Behnken效应面法用于优选肾茶的提取工艺是可行的,建立的数学模型和实验观察数据相符,DM-130大孔吸附树脂较适合肾茶总黄酮的纯化。

板蓝根多糖的提取与脱蛋白工艺优化

通过Box-Behnken组合实验设计,考察了料液比、提取温度及提取时间对板蓝根多糖提取率的影响。利用响应面分析方法,模拟得到二次多项式回归方程的预测模型,并确定最佳提取条件为:料液比为1∶55,提取温度80℃,提取时间3.5 h。在此条件下,板蓝根多糖的提取率为3.55%。以蛋白脱除率和多糖损失率为指标,比较了Sevag法、三氯乙酸法和胃/胰蛋白酶法对粗多糖中蛋白质的脱除效果。结果表明:采用胃/胰蛋白酶脱蛋白工艺,蛋白脱除率较高,分别可达63.5%和52.9%,多糖损失较少,且操作简便,无污染,是较好的脱蛋白方法。

脓毒症合并急性肾损伤患者外周血TLR4、HMGB1、MFG-E8表达水平及临床意义

目的探讨脓毒症合并急性肾损伤患者外周血Toll受体4(TLR4)、高迁移率族蛋白box-1(HMGB1)、乳脂球表皮生长因子-8(MFG-E8)表达水平及临床意义。方法选取本院ICU收治的110例脓毒症患者,其中脓毒症合并急性肾损伤50例(SIAKI组),脓毒症不伴急性肾损伤60例(非AKI组),同时选取同期健康体检正常者50例(对照组),采用单因素方差分析3组患者TLR4、HMGB1、MFG-E8表达差异,采用Pearson分析三者表达相关性,进一步通过Logistic回归分析影响SIAKI严重程度的因素。结果 SIAKI组、非AKI组患者外周血TLR4、HMGB1表达水平明显高于对照组(P <0.05),且SIAKI组高于非AKI组(P <0.05),SIAKI组、非AKI组患者外周血MFG-E8表达水平低于对照组(P <0.05),SIAKI组低于非AKI组(P <0.05);SIAKI组患者C反应蛋白(CRP)、降钙素原(PCT)、血肌酐、尿素、尿酸及急性生理学与慢性健康情况评分系统Ⅱ(APACHEⅡ)评分显著高于非AKI组(P <0.05),肾小球滤过率(eGFR)明显低于非AKI组(P <0.05);SIAKI患者外周血TLR4、HMGB1水平与尿素、尿酸无相关性(P> 0.05),与CRP、PCT、血肌酐、eGFR、APACHEⅡ评分呈正相关(P <0.05);MFG-E8表达水平与CRP、尿素、尿酸无相关性(P>评分呈负相关(P <0.05);CRP、PCT、血肌酐、eGFR、TLR4、HMGB1是影响SIAKI严重程度的危险因素(P <0.05),MFG-E8是影响SIAKI严重程度的保护因素(P <0.05)。结论 SIAKI患者存在外周血TLR4、HMGB1水平升高,MFG-E8水平降低现象,TLR4、HMGB1、MFG-E8可作为SIAKI严重程度的检测指标。

丙泊酚对急性肺损伤大鼠HMGB-1表达的影响

目的探讨丙泊酚的肺保护作用及其可能作用机制。方法采用尾静脉注射内毒素建立急性肺损伤模型,丙泊酚和锌原卟啉同样经过尾静脉给药。将Wistar大鼠按随机数字表法分为实验对照组(C组)、脂多糖组(L组)、脂多糖+丙泊酚组(LP组)。C组尾静脉注射生理盐水;L组尾静脉注射LPS 7.5mg·kg-1;LP组尾静脉注射LPS 7.5mg·kg-1,同时静脉注射丙泊酚30mg·kg-1。给药前及开始后第3、6和12h测定动脉血氧分压(PaO2),实验结束后观察肺湿/干重(W/D)比值、肺组织病理学检查并进行肺损伤轻重程度评分,同时观察各组大鼠肺组织高迁移率族蛋白1(HMGB-1)含量。结果实验前各组PaO2无明显差异,注射LPS后L组PaO2持续下降,和C组相比其PaO2显著降低(P<0.05)。实验结束后L组和C组相比其W/D比值、肺组织病理学检查评分、HO-1及HMGB-1含量显著增加(P<0.05)。而LP组和L组相比其PaO2显著改善,而W/D比值、肺组织病理学检查评分及HMGB-1含量显著降低(P<0.05)。结论丙泊酚具有肺保护作用,且该作用可能与其抑制HMGB-1过度表达作用相关。

黏蛋白1、细胞周期调节蛋白p16和高迁移率族蛋白B1在原发性喉癌临床诊断效能分析

目的评估黏蛋白MUCI(Mucin-1)、p16和高迁移率族蛋白BI(HMGB1)在原发性喉癌中诊断.疾病进展及预后的相关性。方法通过免疫组化方法分析原发性喉癌患者临床肿瘤样本中p16和MUCI表达水平,ELISA法检测原发性喉癌患者血清样本中HMGB1的表达水平。Spearman相关分析MUCI,p16和HMGBI表达水平与喉癌分期、分级的相关性。并通过Kaplan-Meier法分析MUCI,p16和HMGBI表达水平与原发性喉癌患者预后的相关性。结果免疫组化结果示MUCI表达水平与原发性喉癌的分期具有显著相关性(r=0.513,P<0.001),p16与原发性喉癌分期有显著相关性(r=00.437,P<0.001)。HMGBI与喉癌患者的临床分期和病理分级有相关性(r=0.523,r=0.671,P均<0.001)。结论MGBI、p16和MUCI的表达水平状态可做为原发性喉癌患者疾病进展的诊断指标,有望成为原发性喉癌诊断与治疗的新生物标志物。

金丝桃苷双水相提取工艺研究

以鹿蹄草为原料,利用微波双水相辅助提取技术进行提取,在单因素实验的基础上对提取条件进行了考察,根据BBD(Box-Behnken Design)实验设计原理,采用3因素3水平的响应面分析法,以鹿蹄草中金丝桃苷(hy-perin)为指标,对提取过程进行优化,得到最佳工艺参数为:提取温度50℃,液固比36∶1,(NH4)2SO4浓度22%,提取时间22 min,微波功率300 W,提取次数3次。在最佳提取条件下,金丝桃苷的提取率可达0.982 mg.g-1,纯度可以达0.443%。

响应面法优化杨树花多糖超声提取工艺

采用Plackett-Burman设计结合Box-behnken响应面法优化杨树花多糖的超声提取工艺。以杨树花多糖得率为考察指标，对影响杨树花得率的超声功率、超声频率、提取温度、提取时间、提取次数、液料比、颗粒大小、醇沉时间、醇沉浓度等9个因素，采用Plackett-Burman设计从中筛选出具有显著效应的因素一超声功率、提取温度、提取时间；进一步以杨树花多糖得率为考察指标，采用Box-behnken设计响应面法对以上三个因素进行考察，确定最佳工艺条件为：超声功率195W，提取温度66℃，提取时间43min，以此条件提取的多糖含量为63．18±1．57mg／g，与理论预测值仅相差2％～3％。

丙酮酸乙酯对脓毒症大鼠肠组织保护和HMGB1表达的影响

目的:通过观察丙酮酸乙酯对脓毒症大鼠肠组织HMGB1表达的影响,进一步揭示丙酮酸乙酯治疗脓毒症的分子作用机制。方法:将96只SD大鼠随机分为假手术组、脓毒症组、低剂量丙酮酸乙酯治疗组(LET组)、高剂量丙酮酸乙酯治疗组(ET组),以盲肠结扎穿刺法复制大鼠脓毒症模型。LET组(20 mg/kg)及ET组(40 mg/kg)即刻腹腔内注射EP注射液4 mL,每6 h重复注射一次,直至实验结束,假手术组及脓毒症组在相同时间用相同剂量的生理盐水腹腔内注射。各组大鼠分别于术后2 h、8 h、24 h、48 h 4个时间点随机处死6只大鼠,经腹主动脉取血,用ELISA方法检测血浆TNF-α、IL-6、HMGB1水平。术后24 h,取大鼠末端回肠,用RT-PCR法检测回肠组织HMGB1mRNA表达水平,用免疫组织化学两步方法观察肠组织HMGB1蛋白表达,在光镜下观察大鼠肠组织的病理变化。结果:与假手术组相比,脓毒症组血浆HMGB1在术后8 h、24 h、48 h显著增高,脓毒症组回肠组织HMGB1mRNA及蛋白表达在术后24 h显著增高(P<0.05)。与脓毒症组相比,ET组、LET组血浆HMGB1在术后8 h、24 h、48 h显著降低,ET组、LET组回肠组织HMGB1mRNA及蛋白表达在术后24h显著降低(P<0.05)。结论:脓毒症大鼠血浆HMGB1出现时间晚,作用时间长,提示HMGB1是脓毒症的重要晚期炎症介质。丙酮酸乙酯可抑制脓毒症大鼠血浆及肠组织HMGB1的表达,提示丙酮酸乙酯对脓毒症的治疗机制可能与其直接或间接抑制HMGB1的表达有关。

Ethyl pyruvate protects against experimental acute-on-chronic liver failure in rats

AIM： To investigate the protective effects of ethyl py- ruvate （EP） on acute-on-chronic liver failure （ACLF） in rats. METHODS： An ACLF model was established in rats, and animals were randomly divided into normal, mod- el and EP treatment groups. The rats in EP treatment group received EP （40 mg/kg） at 3 h, 6 h, 12 h and 24 h after induction of ACLF. Serum endotoxin, high mobility group box-1 （HMGB1）, alanine transaminase （ALT）, tumor necrosis factor-α （TNF-α）, interferon-α （IFN-γ）, interleukin （IL）-10 and IL-18 levels, changes of liver histology and HMGB1 expressions in liver tis- sues were detected at 48 h after induction of ACLF. The effects of EP on the survival of ACLF rats were also observed.RESULTS： Serum levels of endotoxin （0.394 ± 0.066 EU/mL vs 0.086±0.017 EU/mL, P 〈 0.001）, HMGB1 （35.42±10,86 μg/L vs 2.14 ± 0.27 μg/L, P 〈 0.001）, ALT （8415.87 ± 3567.54 IU/L vs 38.64 ± 8.82 IU/L, P 〈 0.001）, TNF-α （190.77 ± 12.34 ng/L vs 124.40 ± 4.12 ng/L, P 〈 0.001）, IFN-γ （715.38 ± 86.03 ng/L vs 398.66 ± 32.91 ng/L, P 〈 0.001）, IL-10 （6.85 ± 0.64 ng/L vs 3.49 ± 0.24 ng/L, P 〈 0.001） and IL-18 （85.19 ±3.49 ng/L vs 55.38 ±1.25 ng/L, P 〈 0.001） were significantly increased, and liver tissues presented se- vere pathological injury in the model group compared with the normal group, Howeverr EP administration significantly improved hepatic histopathology and re- duced the serum levels of endotoxin （0.155±0.045 EU/mL vs 0.394 ± 0.066 EU/mL vs P 〈 0.001） and in- flammatory cytokines （11.13 ± 2.58 μg/L vs 35.42 ± 10.86 μg/L for HMGB1, 3512.86 ± 972.67 IU/L vs 8415.87 ± 3567.54 IU/L for ALT, 128.55 ± 5.76 ng/L vs 190.77 ± 12.34 ng/L for TNF-α 438.16 ± 38.10 ng/L vs 715.38 ± 86.03 ng/L for IFN-γ 3.55 ± 0.36 ng/L vs 6.85 ± 0.64 ng/L for IL-10, and 60.35 ± 1.63 ng/L vs 85.19 ± 3.49 ng/L for IL-18, respectively, P 〈 0.001）, and the levels of HMGB1 in liver tissues re- gardless of treatment time after induction of ACLF. EP treatment at the four time points prolonged the me- dian survival time of ACLF rats （60 h） to 162 h, 120 h, 102 h and 78 h, respectively （χ2 = 41.17, P 〈 0.0001）. CONCLUSION： EP administration can protect against ACLF in rats, and is a potential and novel therapeutic agent for severe liver injury.

Glycyrrhizic acid promotes sciatic nerves recovery in type 1 diabetic rats and protects Schwann cells from high glucose-induced cytotoxicity

The present study aims to investigate the therapeutic effect and mechanism of glycyrrhizic acid(GA)in diabetic peripheral neuropathy(DPN).GA significantly mitigated nerve conduction velocity(NCV)deficit and morphological abnormality and reduced high-mobility group box-1(HMGB1)expression in the sciatic nerves of diabetic rats independent of blood glucose and body weight.Notably,GA alleviated the increase of HMGB1 and the decrease of cell viability in high glucose-stimulated RSC96 cells.Furthermore,GA obviously reduced the concentration of inflammatory cytokines in the sciatic nerves of diabetic rats and supernatants of high glucose-exposed RSC96 cells,then restored the decreased expression levels of nerve growth factor(NGF)and neuritin-1,and the increased expression levels of cleaved caspase-3 and neuron-specific enolase.Additionally,GA markedly inhibited receptor for advanced glycation end products(RAGE)expression,p38MAPK phosphorylation,and the nuclear translocation of NF-κBp65 in diabetic rats and high glucose-exposed RSC96 cells.The promotional effect of high glucose in RSC96 cells was diminished following Hmgb1 siRNA treatment.Our findings indicate that GA may exert neuroprotection on DPN by suppressing HMGB1,which lead to extenuation of inflammation response,balance of NGF,neuritin-1 and caspase-3,as well as inactivation of RAGE/p38MAPK/NF-κBp65 signaling pathway.

High mobility group box 1 protein(HMGB1)as an immune-modulating factor for polarization of human T lymphocytes

Objective This study was performed to investigate the effect of high mobility group box-1 protein(HMGB1)on immune function of human T lymphocytes in vitro and explore its potential role in cell-mediated immune dysfunction.Methods Fresh blood was obtained from healthy adult volunteers and peripheral blood mononuclear cells(PBMCs)were isolated,then rhHMGB1 was added to PBMCs.Four-color flow cytometric(FCM)analysis was used for the measurement of intracellular cytokine including interleukin IL-4 and interferon IFN-?ELISA kits were employed for the determination of IL-2 and sIL-2R protein levels in cell culture supernatants.Results(1)Different stimulating time and dosage of rhHMGB1 did not alter the number of IFN-αpositive cells(Th1).rhHMGB1 stimulation provoked a dose-dependent and time-dependent increase in Th2 subset and decrease in ratio of Th1 to Th2.(2)Compared with the untreated cells,when the cells were coincubated with rhHMGB1(10-100ng/ml)for 12 hrs,protein release of IL-2 and sIL-2R were significantly up-regulated.At 48 hrs,in contrast,protein production was relatively lower in cells after exposure to 100-1000 ng/ml rhHMGB1.Conclusions These findings demonstrated that HMGB1 has a dual influence on immune functions of human T lymphocytes.

Sodium butyrate protects against toxin-induced acute liver failure in rats

BACKGROUND: Acute liver failure(ALF) is a serious clinical syndrome with high mortality. Sodium butyrate has been shown to alleviate organ injury in a wide variety of preclinical models of critical diseases. The aim of this study was to investigate the protective effect of sodium butyrate on ALF in rats.METHODS: All rats were randomly divided into control,model and sodium butyrate treatment groups. Except the control group, the rats were induced ALF animal model by subcutaneous injection of human serum albumin+D- galactosamine+lipopolysaccharide. After induction of ALF,the rats in the treatment group received sodium butyrate(500mg/kg) at 12-hour or 24-hour time point. Fourty-eight hours after ALF induction, the animals were sacrificed and samples were harvested. Serum endotoxin, high mobility group box-1(HMGB1), liver function parameters, tumor necrosis factoralpha(TNF-α) and interferon-gamma(IFN-γ) were measured.The expression of HMGB1 and nuclear factor-kappa B(NF-κB)p65 protein in liver tissue was detected by Western blotting. The histological changes of liver and intestine were examined. The survival duration was also observed.RESULTS: Serum endotoxin, alanine aminotransferase, HMGB1,TNF-α and IFN-γ were significantly increased and the liver histology showed more severe histopathological injury in the model group compared with the control group(P<0.05).Compared to the model group, sodium butyrate treatment significantly improved the histopathological changes in the liver and intestine, reduced serum endotoxin and inflammatory cytokines, suppressed HMGB1 and NF-кB p65 proteins in liver tissue, and prolonged the survival duration regardless of treatment at 12 hours or 24 hours after induction of ALF(P<0.05).CONCLUSIONS: Sodium butyrate protected the liver from toxin-induced ALF in rats. The mechanisms may be due to direct hepatoprotection and decreased intestinal permeability.