硫化氢与糖尿病

硫化氢是继一氧化氮和一氧化碳之后的第三种内源性气体信号分子。近年来,内源性硫化氢的产生及生理意义已经被认识,其代谢异常与多种疾病有关。本文综述了近年报道的硫化氢及其内生酶在糖尿病发病及进展中的变化情况,并重点概述硫化氢效应的细胞机制,包括硫化氢对β细胞胰岛素释放和对脂肪细胞葡萄糖摄取的影响。深入理解硫化氢在糖尿病中的作用将为以硫化氢为靶点的糖尿病治疗和抗糖尿病新药设计提供新的思路。     

硫化氢与细胞的增殖和凋亡

硫化氢是内源性气体分子家族中的一员,是一种气体递质（gasotransmitter）。近年来,内源性硫化氢的产生及生理意义已经被认识,其代谢异常与许多疾病有关。本文综述了最近发现的硫化氢对细胞增殖和凋亡的调节作用,并重点概述硫化氢细胞效应的分子机制,包括丝裂原活化蛋白激酶、细胞周期相关激酶、细胞死亡相关基因以及离子通道等的改变。对硫化氢调节细胞生长或死亡的深入了解将为新药设计及许多疾病的治疗提供新的思路。     

内源性硫化氢体系与低O2高CO2性肺动脉高压的相关性研究

目的：研究低O2高CO2性肺动脉高压（HHPH）时大鼠肺组织内硫化氢（H2S）体系变化的改变及相关性,并探讨其机制。方法：20只SD大鼠随机分为正常对照组（C组）和低O2高CO2组（HH组）（n=10）。监测其血流动力学变化,光镜观察肺细小动脉管壁面积／管总面积比值（WA／TA）,测右心室／左心室＋室间隔（RV／LV＋SP）比值。原位缺口末端标记法（TUNEL法）观测肺细小动脉凋亡情况,并计算凋亡指数（AI）,免疫组化检测肺细小动脉Bcl-2、Bax蛋白表达。敏感硫电极法测定血浆H2S含量及肺组织匀浆胱硫醚-γ-裂解酶（CSE）活性变化。RT-PCR法测定肺组织中CSE基因表达。结果：HH组肺平均动脉压（mPAP）、WA／TA、RV／LV＋SP明显高于C组（P〈0．05或P〈0．01）。与C组相比,HH组AI显著下降（P〈0．01）,Bcl-2表达加强,Bax减弱,Bax／Bcl-2比值上升（均P〈0．01）。血浆H2S含量、肺组织匀浆CSE活性、肺组织CSE基因表达水平HH组明显低于C组（P〈0．01）。血浆H2S、肺组织CSE活性、肺组织CSE mRNA相对含量与mPAP,Bcl-2／Bax比值均呈显著负相关,与AI呈显著正相关。结论：H2S／CSE体系与低O2高CO2性肺动脉高压关系密切,HHPH时H2S／CSE体系的明显受抑,可使Bcl-2／Bax比值升高,肺动脉平滑肌细胞凋亡减少,促进肺血管重建和肺动脉高压的形成。     

去势大鼠阴茎海绵体胱硫醚γ裂解酶与硫化氢的表达

目的观察去势大鼠阴茎海绵体胱硫醚γ裂解酶（Cystathionine-γ-lyase,CSE）/硫化氢（Hydrogen sulphide,H2S）的变化,进一步探讨勃起功能障碍的发病机制。方法雄性SD大鼠72只分4组：对照组、假手术组,去势组和去势炔丙基甘氨酸（PAG,CSE阻断剂）组,检测基础条件下和阿扑吗啡（Apomorphine,APO）刺激后的海绵体内压（Intracavernous pressure,ICP）及勃起率;激光共聚焦显微镜检测CSE在大鼠勃起不同时期阴茎海绵体组织中的表达,敏感硫电极测定H2S在勃起不同时期的含量。结果与假手术组比较,去势组和去势PAG组ICP与勃起率下降（P〈0.01）;且去势PAG组较去势组ICP明显下降（P〈0.01）;阿朴吗啡刺激后,与假手术组比较,勃起前去势组和去势PAG组CSE蛋白表达降低（P〈0.01）,勃起中去势各组较假手术组CSE蛋白表达降低（P〈0.01）,且去势PAG组较去势组CSE蛋白表达明显降低（P〈0.01）;勃起后各组间CSE蛋白表达变化无差异。勃起前和勃起中去势各组较假手术组H2S含量下降（P〈0.05）,且勃起中去势PAG组较去势组H2S含量明显下降（P〈0.01）：勃起后去势组和去势PAG组较假手术组H2S含量下降（P〈0.01）。结论去势大鼠勃起功能障碍与CSE和H2S表达下降有关。     

内、外源性硫化氢在脂多糖所致大鼠急性肺损伤中的作用

目的：探讨内、外源性硫化氢（H2S）在脂多糖（LPS）所致大鼠急性肺损伤（ALI）中的作用并初探其机制。方法：将120只SD大鼠随机分为对照组、IPS组（经气管内滴注LPS复制ALI模型）、NaHS＋LPS组和炔丙基甘氨酸（PPG）＋LPS组。给药后4h或8h处死动物,测定肺系数;光镜观察肺组织形态学改变;化学法检测血浆H2S、NO和CO含量、肺组织丙二醛（MDA）含量、胱硫醚-γ-裂解酶（CSE）、诱导型一氧化氮合酶（iNOS）和血红素加氧酶（HO）活性以及支气管肺泡灌洗液（BALF）中中性粒细胞（PMN）数目和蛋白含量的变化;用免疫组织化学法检测肺组织iNOS、HO-1蛋白表达。再将血浆H2S含量与上述指标进行相关性分析。结果：气管内滴注LPS可引起肺组织明显的形态学改变;肺系数和肺组织MDA含量增加;BALF中PMN数目和蛋白含量增加;血浆H2S含量和肺组织CSE活性下降;肺组织iNOS活性、HO活性和iNOS蛋白表达、HO-1蛋白表达增强,血浆NO含量、CO含量增加。预先给予NaHS可显著减轻LPS所致上述指标的改变;而预先给予PIG可加重LPS所致肺损伤,使BALF中PMN数目和蛋白含量、血浆NO含量、肺组织iNOS活性和iNOS蛋白表达进一步增加,但对血浆CO含量、肺组织HO活性和HO-1蛋白表达无明显影响。HS含量与CSE活性、血浆CO含量、肺组织HO-1活性呈正相关（r值=0．945—0．987,P均〈0．01）;与其他指标呈负相关（r值=-0．994～-0．943,P均〈0．01）。结论：H2S／CSE体系的下调在LPS所致大鼠Ⅲ的发病学中有一定作用,内、外源性H2S具有抗LPS所致Au的作用,该作用可能与其抗氧化效应、减轻PMN所致肺过度的炎症反应以及下调NO／iNOS体系、上调CO／HO—1体系有一定关系。     

大鼠肝硬化形成中气体信号分子一氧化氮、一氧化碳对硫化氢/胱硫醚-γ-裂解酶体系的影响

目的:研究气体信号分子一氧化氮(NO)、一氧化碳(CO)在大鼠肝硬化中对硫化氢(H2S)/胱硫酶-γ-裂解酶(CSE)体系的影响.方法:将SD大鼠随机分为3组:正常对照组(NC组)、肝硬化模型组(CIRM组)、肝硬化模型+左旋硝基精氨酸甲酯(L-NAME)+锌原卟啉-Ⅸ(Znpp-Ⅸ)组(CIRM+L-NAME+ZnPP-Ⅸ组).采用复合因素法建立大鼠肝硬化模型.运用敏感硫电极方法检测大鼠血浆中H2S的含量;应用免疫组织化学方法对大鼠门静脉(PV)上CSE蛋白进行定位及半定量分析;利用蛋白质免疫印迹技术(Western blot)4定大鼠肝组织中CSE蛋白的表达量.结果:在大鼠肝硬化形成后,血浆H2S含量显著降低,CIRM组[(134.49±12.25)umol/L]与NC组[(180.34±11.71)umol/L]比较有显著性差异(P<0.01);门静脉上CSE蛋白的表达[NC组、CIRM组分别为(139.19±1.13)、(153.68±0.90)]及其在肝组织中的蛋白含量[NC组、CIRM组分别为(159.30±1.37)、(121.72±1.61))显著降低(P<0.01);当外源性给予L-NAME、ZnPP-Ⅸ后,CIRM+LNAME+ZnPP-Ⅸ组血浆H2S含量[(160.81±6.79)u-mol/L]显著高于CIRM组[(134.49±12.25)umol/L,P<0.01],同时,CIRM+L-NAME+ZnPP-Ⅸ组门静脉上CSE蛋白的表达(148.72±1.39)及其在肝组织中的蛋白含量(142.79±1.13)显著高于CIRM组(P<0.01).结论:在大鼠肝硬化形成中,NO、CO可以上调H2S/CSE体系.     

NO前体对高肺血流时肺动脉胱硫醚-γ-裂解酶基因表达的调节作用

目的：研究一氧化氮（NO）前体L-精氨酸（L—Arg）对高肺血流时肺动脉胱硫醚-γ-裂解酶（内源性硫化氢生成酶）的调节作用,以探讨NO体系对高肺血流肺动脉高压及肺血管结构重建调节作用的机制。方法：30只雄性SD大鼠随机分为对照组,分流组和分流＋L—Arg组。对后两组大鼠行腹主动脉-下腔静脉分流术。观察术后11周大鼠肺动脉平均压（mPAP）、右心室肥厚和肺动脉相对中膜面积的改变,用竞争逆转录聚合酶链反应（RT-PCR）对肺组织CSEmRNA表达进行定量分析,同时用化学法测定肺组织硫化氢产出率。结果：分流组大鼠mPAP及肺动脉相对中膜面积明显高于对照组（P〈0．01）,而分流＋L—Arg组大鼠mPAP及肺动脉相对中膜面积明显低于分流组（P〈0．01）。分流组CSEmRNA表达与对照组相比明显降低（P〈0．01）,而分流组＋L—Arg组CSEmRNA表达又明显高于分流组（P〈0．05）：分流组大鼠肺组织硫化氢产出率明显低于对照组（P〈0．01）,而分流组＋L—Arg组大鼠肺组织硫化氢产出率及血浆硫化氢含量明显高于分流组（P〈0．01）。结论：高肺血流可致肺动脉CSEmRNA下调,外源性NO能够缓解CSEmRNA的改变,从而对高肺血流所致肺血管结构重建和肺动脉高压起调节作用。     

脂肪因子Chemerin与妊娠相关疾病的关系展望

<正>1 Introduction Recurrent pregnant loss,gestational diabetes,premature delivery,intrauterine growth restriction,preeclampsia and other pregnancy-related complications have severe impact on the fetus development and the health and life quality of the mother.These diseases are also causes of unstability and huge economic burden for the family as well as the     

人胱硫醚β-合酶及其截短型片段的表达、纯化和活性测定

将人胱硫醚β-合酶(CBS)基因克隆至质粒pGEX-4T-1中,获得的重组质粒pGEX-4T-1-CBS转入大肠杆菌E.coli Rosetta (DE3)菌株,构建了高效表达CBS的重组菌E.coli Rosetta (pGEX4T-1-CBS)。重组菌在0.1mmol/L的IPTG于30℃诱导16h,可溶性CBS表达量达到28mg/L培养基。将重组菌破碎后上清液经GSTrap Fast Flow亲和层析一步纯化得到CBS融合蛋白,在凝血酶柱上切割缓冲液中加入3%甘油和0.1%CHAPS可以有效抑制酶切后CBS聚沉,酶活性回收率为54.8%,蛋白质产率为15.2mg/L培养基,纯度达到95%,单位酶活为143U/mg,终浓度为1mmol/L的S-腺苷甲硫氨酸(AdoMet)可使CBS单位酶活提高5.1倍,达到735U/mg。同时构建了表达CBS_1-413(删除了CBS羧基端调控域138个氨基酸残基)的重组菌E.coli Rosetta (pETDuet-1-CBS_1-413),经过一步HisTrap Fast Flow亲和层析,酶活性回收率为74.3%,蛋白质产率为12.8mg/L培养基,纯度达到95%,单位酶活为965U/mg; 还表达和纯化了胱硫醚β-裂解酶(CBL),并在此基础上建立了一种新的CBL偶联的CBS酶活性测定方法。     

内源性气体硫化氢在中枢神经系统中的作用

内源性硫化氢(H2S)可以刺激神经细胞cAMP水平增加,提高NMDA受体介导的突触后兴奋性电位,提高诱导海马长时程增强。H2S不仅具有神经调节剂的功能,还有神经保护剂的功能。H2S自身并不能将细胞从氧化应激中解救出来,但是它能通过提高胞内有效的抗氧化剂——还原型谷胱苷肽的含量而起到保护神经元的作用。对H2S的研究刚刚起步,对其在神经系统中的作用机制开展研究将有助于了解其在神经元保护方面所起的作用。     

Hydrogen Sulfide and Carbon Monoxide Are in Synergy with Each Other in the Pathogenesis of Recurrent Febrile Seizures

1. The purpose of the present study was to investigate the interaction between hydrogen sulfide (H(2)S) and carbon monoxide (CO) during recurrent febrile seizures (FS) 2.H(2)S and CO are important intra- and intercellular messengers, regulating various brain functions. Our recent studies showed that both of them alleviate the hippocampal damage induced by recurrent FS. In the present study, on a rat model of recurrent FS, we found that hydroxylamine (an inhibitor of cystathionine b-synthase, CBS) reduced CO level and down regulated heme oxygenase (HO-1) expression, while NaHS (a donor of H(2)S) elevated CO level and upregulated HO-1 expression. ZnPP-IX (an inhibitor of HO-1) decreased H(2)S formation and down regulated CBS expression, while hemin (which increases the production of endogenous CO) enhanced H(2)S formation and elevated CBS expression. 3.Our data demonstrate that endogenous H(2)S and CO are in synergy with each other in recurrent FS.     

Hydrogen sulfide from adipose tissue is a novel insulin resistance regulator

Recent data suggested that endogenous hydrogen sulfide (H₂S) contributes to the pathogenesis of diabetes. Here, we identified that cystathionine gamma lyase (CSE) was expressed in adipose tissue in rats and endogenously generated H₂S. The CSE/H₂S system exists in both rat adipocytes and pre-adipocytes. This system was up-regulated with aging, although a high level of glucose down-regulated the system in a concentration- and time-dependent manner. H₂S inhibited the basal and insulin-stimulated glucose uptake of mature adipocytes, whereas administration of CSE inhibitors enhanced the glucose uptake of adipocytes. The PI3K but not K_ATP channel pathway is involved in the inhibitory effect of H₂S on glucose uptake. Finally, in fructose-induced diabetes in rats, we confirmed the up-regulated CSE/H₂S system in adipose tissue, which was negatively correlated with glucose uptake in this tissue. Our findings suggest that H₂S might be a novel insulin resistance regulator.     

Endogenous CSE/H2S system mediates TNF‐α‐induced insulin resistance in 3T3‐L1 adipocytes

Tumour necrosis factor‐α (TNF‐ α)is a major contributor to the pathogenesis of insulin resistance associated with obesity and type 2 diabetes. It has been found that endogenous hydrogen sulfide (H₂S) contributes to the pathogenesis of diabetes. We have hypothesized that TNF‐α‐induced insulin resistance is involved in endogenous H₂S generation. The aim of the present study is to investigate the role of endogenous H₂S in TNF‐α‐induced insulin resistance by studying 3T3‐L1 adipocytes. We found that treatment of 3T3‐L1 adipocytes with TNF‐α leads to deficiency in insulin‐stimulated glucose consumption and uptake and increase in endogenous H₂S generation. We show that cystathionine γ‐lyase (CSE) is catalysed in 3T3‐L1 adipocytes to generate H₂S and that CSE expression and activity are upregulated by TNF‐α treatment. Inhibited CSE by its potent inhibitors significantly attenuates TNF‐α‐induced insulin resistance in 3T3‐L1 adipocytes, whereas H₂S treatment of 3T3‐L1 adipocytes impairs insulin‐stimulated glucose consumption and uptake. These data indicate that endogenous CSE/H₂S system contributes to TNF‐α‐caused insulin resistance in 3T3‐L1 adipocytes. Our findings suggest that modulation of CSE/H₂S system is a potential therapeutic avenue for insulin resistance. Copyright © 2012 John Wiley & Sons, Ltd.     

β-Cyclodextrin-Based Nitrosoglutathione Improves the Storage Quality of Peach by Regulating the Metabolism of Endogenous Nitric Oxide,Hydrogen Sulfide,and Phenylpropane

Nitrosoglutathione (GSNO) and β-cyclodextrin (β-CD) exhibit positive roles in regulating fruit quality. However, there are few reports about the effects of GSNO and β-CD on enhancing storability and boosting nitric oxide (NO), hydrogen sulfide (H₂S), and phenylpropane metabolism in fruits during storage. “Xintaihong” peach were treated with 0.5, 1.0, 1.5 mmol L⁻¹ GSNO in 0.5% (w/v) β-CD solution (GSNO/β-CD). The effects of GSNO/β-CD on endogenous NO, H₂S, and phenylpropane metabolism were investigated. Treatment with GSNO/β-CD increased the color difference of peach and inhibited the increase of respiratory intensity, weight loss, and relative conductivity. Treatment with 1.0 mmol L⁻¹ GSNO/β-CD increased the nitric oxide synthase (NOS-like) activity and L-arginine content, thereby promoting the accumulation of endogenous NO. By improving the activities of L-cysteine desulfhydrylase (L-CD), O-acetylserine sulfur lyase (OAS-TL), serine acetyltransferase (SAT), GSNO/β-CD increased the content of endogenous H₂S in peach. Treatment with GSNO/β-CD increased the activities of phenylalanine ammonia-lyase (PAL), 4-coumarate-CoA ligase (4CL), and cinnamic acid-4-hydroxylase (C4H), promoted the increase of total phenols, flavonoids, and lignin in peach. These results indicated that GSNO/β-CD treatment better maintained the quality of peach by improving the metabolism of endogenous NO, H₂S, and phenylpropane during storage.     

硫化氢在糖尿病及其并发症中作用的研究进展

糖尿病及其并发症是覆盖全球的常见疾病,发病率逐年增高。硫化氢是继一氧化氮和一氧化碳之后的第三种气体信号分子,发挥重要病理生理学效应。目前研究发现,硫化氢在调节胰岛β细胞功能、胰岛素抵抗和糖尿病并发症中发挥着重要作用,已经成为糖尿病及其并发症的研究热点。本篇综述就H_2S在糖尿病及其并发症中发挥的病理生理学作用及机制进行了阐述。     

胆汁酸对代谢性疾病的调控

胆汁酸作为一种信号分子通过激活肝、肠道和外周组织中的胆汁酸受体影响体内葡萄糖和脂质的代谢平衡,对于调节肥胖、2型糖尿病和非酒精性脂肪肝等代谢性疾病具有非常重要的意义。胆汁酸与相应核受体,如法尼醇X受体(farnesoid X receptor, FXR)和Takeda G蛋白偶联受体5 (Takeda G protein-coupled receptor 5,TGR5)的相互作用影响了这些代谢性疾病。FXR主要通过影响胆汁酸的合成及转运对非酒精性脂肪肝发挥作用,TGR5则是间接增加褐色脂肪组织中的生热作用,改善肥胖和2型糖尿病。这些调控机制的研究是非常必要的。本文综述了胆汁酸代谢及其对代谢性疾病调控的分子机制的研究进展,以期为科研工作者提供一定的参考。