Programmed cell death, antioxidant response and oxidative stress in wheat f lag leaves induced by chemical hybridization agent SQ-1

Male sterility induced by a chemical hybridization agent （CHA） is an important tool for utilizing crop heterosis. Leaves, especially the flag leaves, as CHA initial recipients play a decisive role in inducing male sterility. To investigate effects of different treatment times of CHA-SQ-1 used, morphological, biochemical and physiological responses of wheat flag leaves were detected in thistudy. CHA induced programmed cell death （PCD） as shown in terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling （TUNEL） and DNA laddering analysis. In the early phase, CHA-SQ-1 trig- gered organelle changes arid PCD in wheat leaves accompanied by excess production of reactive oxygen species （O2- and H202） and down-regulation of the activities of superoxide dismutase （SOD）, catalase （CAT） and guaiacol peroxidase （POD）. Meanwhile, leaf cell DNAs showed ladder-like patterns on agarose gel, indicating that CHA-SQ-1 led to the activation of the responsible endonuclease. The oxidative stress assays showed that lipid peroxidation was strongly activated and photosynthesis was obviously inhibited in SQ-l-induced leaves. However, CHA contents in wheat leaves gradually reduced along with the time CHA-SQ-1 applied. Young flags returned to an oxidative/antioxidative balance and ultimately developed into mature green leaves. These results provide explanation of the relations between PCD and anther abortion and practical application of CHA for hybrid breeding.

氧化苦参碱水凝胶通过激活角化细胞Nrf2/HO-1通路促进创面愈合

背景:慢性创面炎症和氧化应激阻碍了角化细胞的再生,氧化苦参碱具有抗氧化、抗炎等多种生物活性,可能具有促进创面愈合的潜在效应。目的:探讨氧化苦参碱对创面愈合的作用,以及对H_(2)O_(2)诱导人角质形成细胞系HaCaT细胞氧化应激损伤的保护作用。方法:①体内实验:分别制备含0,0.05,0.1,0.2 g/L氧化苦参碱的甲基丙烯酰化透明质酸(HAMA)水凝胶。在75只糖尿病小鼠背中部制作直径12 mm的全层皮肤缺损模型,随机分5组干预,每组15只:模型组创面包扎固定,单纯水凝胶组以HAMA水凝胶覆盖创面,低、中、高剂量氧化苦参碱组分别以含0.05,0.1,0.2 g/L氧化苦参碱的HAMA水凝胶覆盖创面,光固化后包扎固定,14 d内进行相关指标的检测。②体外实验:将人角质形成细胞系HaCaT分5组培养,正常组常规培养,H_(2)O_(2)组及低、中、高浓度氧化苦参碱组均加入H_(2)O_(2)干预4 h,然后分别更换为含0,0.05,0.1,0.2 g/L氧化苦参碱的培养基,培养24 h后进行相关指标的检测。结果与结论:①体内实验:与模型组比较,单纯水凝胶组小鼠创面愈合率无明显变化,低、中、高剂量氧化苦参碱组治疗后7,14 d的创面愈合率升高(P<0.05)。治疗后14 d的创面样本切片病理观察显示,与模型组相比,中、高剂量氧化苦参碱组再生表皮层厚度、显微血管数量与胶原沉积均增加(P<0.05)。术后7 d的创面样本Western blotting检测显示,与模型组相比,中、高剂量氧化苦参碱组肿瘤坏死因子α与白细胞介素6的蛋白表达降低(P<0.05)。②体外实验:CCK-8检测、EdU及Ki67染色显示,与H_(2)O_(2)组相比,中、高浓度氧化苦参碱组细胞增殖能力明显升高(P<0.05)。与H_(2)O_(2)组相比,中、高浓度氧化苦参碱组线粒体膜电位升高(P<0.05)、活性氧含量降低(P<0.05)。Western blotting检测显示,与H_(2)O_(2)组相比,高浓度氧化苦参碱组Nrf2核蛋白、Nrf2总蛋白、HO-1蛋白及超氧化物歧化酶1蛋白的表达增加(P<0.05)。③结果表明:氧化苦参碱能够通过上调Nrf2、HO-1蛋白减轻HaCat细胞的氧化应激损伤,加速创面愈合。

刺五加新品种‘紫加1号’多糖分离纯化及抗氧化活性分析

‘紫加1号’为该课题组选育的刺五加(Acanthopanax senticosus)新品种,嫩茎叶灰紫色且味甘。为分离纯化‘紫加1号’嫩茎叶多糖,并测定各组分的单糖组成和分子量大小、评价各组分的抗氧化活性。该研究以‘紫加1号’嫩茎叶为材料,采用水提醇沉工艺,经大孔树脂吸附法除杂脱色得到粗多糖(A.polysaccharides,ASPS);通过DEAE-Cellulose 52离子柱和Sephadex G-100凝胶柱分离纯化得多糖组分;采用离子色谱和凝胶色谱-示差-多角度激光光散射色谱,测定各均一组分的单糖组成和分子量;通过测定均一组分清除羟基自由基(·OH)、超氧阴离子自由基(O_(2)^(-·))、1,1-二苯基-2-三硝基苯肼自由基(DPPH)的能力,分析其体外抗氧化活性强弱。结果表明:ASPS经分离纯化得到4种多糖组分,即酸性多糖ASPA-1-1、ASPA-2-1、ASPA-3-1和中性多糖ASPN-1,其分子量依次为8.10、26.15、0.91、0.89 kDa,主要是由阿拉伯糖、鼠李糖、半乳糖、葡萄糖、木糖、甘露糖、核糖、半乳糖醛酸和葡萄糖醛酸等按不同比例组成的杂多糖。ASPA-1-1、ASPA-2-1、ASPA-3-1、ASPN-1均具有明显的抗氧化活性,其中ASPA-2-1清除·OH、DPPH的能力高于ASPA-1-1、ASPA-3-1、ASPN-1,而ASPA-3-1清除O_(2)^(-·)的能力最强。综上认为,从‘紫加1号’中分离纯化得到的多糖具有较好的抗氧化活性。该研究结果为‘紫加1号’作为天然抗氧化剂的深度研究和进一步开发与利用提供了一定的科学理论依据。

植物乳杆菌1-1-2发酵去除葡萄柚果汁苦味物质及其抗氧化活性

通过筛选功能性寡糖代谢较强的菌株发酵葡萄柚汁有潜力对其中的柚皮苷进行水解,从而去除苦味。结果显示,通过植物乳杆菌1-1-2发酵后果汁中柚皮苷含量降低,同时,柚皮素含量显著提高,总酚含量和1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除率也显著提高。DPPH自由基清除活性与总酚含量和柚皮素含量呈正相关。植物乳杆菌1-1-2可将柚皮苷进行水解,转化为没有苦味的柚皮素,而柚皮素的抗氧化活性显著高于柚皮苷。发酵后的葡萄柚汁感官评价得分高于未发酵葡萄柚汁。本研究可为改善葡萄柚口感、开发活性更强的功能性益生菌发酵产品提供理论依据。

S-adenosyl-L-methionine modifies antioxidant-enzymes,glutathione-biosynthesis and methionine adenosyltransferases-1/2 in hepatitis C virus-expressing cells

AIM: To elucidate the mechanism(s) by which S-adenosyl-L-methionine(SAM) decreases hepatitis C virus(HCV) expression.METHODS: We examined the effects of SAM on viral expression using an HCV subgenomic replicon cell culture system. Huh7 HCV-replicon cells were treated with 1 mmol/L SAM for different times(24-72 h), then total RNA and proteins were isolated. c DNA was synthesized and real time-PCR was achieved to quantify HCV-RNA, superoxide dismutase 1 and 2(SOD-1, SOD-2) catalase, thioredoxin 1, methionine adenosyltransferase 1A and 2A(MAT1A, MAT2A) expression, and GAPDH and RPS18 as endogenous genes. Expression of cellular and viral protein was evaluated by western-blot analysis using antibodies vs HCV-NS5 A, SOD-1, SOD-2, catalase, thioredoxin-1, MAT1 A, MAT2 A, GAPDH and actin. Total glutathione levels were measured at different times by Ellman's recycling method(0-24 h). Reactive oxidative species(ROS) levels were quantified by the dichlorofluorescein assay(0-48 h); Pyrrolidin dithiocarbamate(PDTC) was tested as an antioxidant control and H2O2 as a positive oxidant agent.RESULTS: SAM exposition decreased HCV-RNA levels 50%-70% compared to non-treated controls(24-72 h). SAM induced a synergic antiviral effect with standard IFN treatment but it was independent of IFN signaling. In addition, 1 mmol/L SAM exposition did not modify viral RNA stability, but it needs cellular translation machinery in order to decrease HCV expression. Total glutathione levels increased upon SAM treatment in HCV-replicon cells. Transcriptional antioxidant enzyme expression(SOD-1, SOD-2 and thioredoxin-1) was increased at different times but interestingly, there was no significant change in ROS levels upon SAM treatment, contrary to what was detected with PDTC treatment, where an average 40% reduction was observed in exposed cells. There was a turnover from MAT1A/MAT2 A, since MAT1 A expression was increased(2.5 fold-times at 48 h) and MAT2 A was diminished(from 24 h) upon SAM treatment at both the transcriptional and translational level. CONCLUSION: A likely mechanism(s) by which SAM diminish HCV expression could involve modulating antioxidant enzymes, restoring biosynthesis of glutathione and switching MAT1/MAT2 turnover in HCV expressing cells.

褪黑素通过SIRT1-BMAL1通路减轻神经病理性疼痛的机制研究

目的评价褪黑素对神经病理性疼痛夜间加重的影响,并通过特异性沉默信息调节因子1(SIRT1)-脑和肌肉ARNT样蛋白1(BMAL1)通路探讨其机制。方法96只SPF级雄性C57/B6小鼠,随机分为3组:假手术(S)组、神经病理性疼痛模型(NP)组和NP模型+褪黑素治疗(NP+M)组;术前试验小鼠置于指定光照模式环境中;采用12 h光照与12 h黑暗交替环境,持续至少两周,将自然时间转换为授时时间(ZT),光照起点定为ZT0;S组小鼠仅分离坐骨神经,NP组和NP+M组采用坐骨神经慢性缩窄性损伤制备小鼠NP模型,NP+M组术后注射10 mg/kg褪黑素;Western blot法检测术后14 d各时点脊髓的SIRT1、BMAL1和谷胱甘肽过氧化酶1(Gpx1)表达量的变化;术后通过免疫荧光技术对脊髓背角SIRT1和脊髓神经元标志物神经元特异性核蛋白(NeuN)、小胶质细胞激活标记物离子钙结合适配器分子1(iba-1)、星形胶质细胞标记物胶质纤维酸性蛋白(GFAP)进行共染色,并检测各时点iba-1以确定小胶质细胞激活状态。结果与NP组ZT10时点相比,NP组小鼠术后14 d ZT22时点SIRT1、BMAL1和Gpx1降低(P<0.05);与NP组ZT14时点相比,NP+M组ZT14时点SIRT1与BMAL1升高(P<0.05),而Gpx1于ZT18时点升高(P<0.05)。SIRT1在脊髓背角与小胶质细胞共表达;与ZT10时点相比,NP组小鼠术后14 d ZT22时点小胶质细胞表达降低(P<0.05);与ZT10时点相比,NP+M组小鼠术后14 d ZT22时点小胶质细胞表达差异无统计学意义。结论褪黑素可以减轻神经病理性疼痛夜间加重,其机制可能与激活小胶质细胞SIRT1-BMAL1通路蛋白表达有关。

New antioxidant SkQ1 is an effective protector of rat eye retinal pigment epithelium and choroid under conditions of long-term organotypic cultivation

Cells have intrinsic mechanisms for cleaning harmful oxidants represented mainly by reactive oxygen species (ROS). Despite the antioxidant defense, ROS can cause serious damage to the retina that with age leads to various eye diseases and even blindness. Among numerous cell sites of ROS generation, mitochondrial electron transport is of crucial importance. Recently, for the purpose of cleaning ROS in the mitochondrial matrix, powerful mitochondria- targeted antioxidant “SkQ1” has been invented. We studied SkQ1 effects upon tissues of rat posterior eye cup that consisted: retinal pigment epithelium (RPE) ? choroidal coat ? scleral coat. The eye cups were isolated from the eyes of adult albino rats and cultivated in rotary tissue culture system in the presence of 20 nM SkQ1 or without this compound. After 7 days - 1 month in vitro eye cup samples were studied by immunohistochemistry, routine histology, morphometry, and digital image analysis. We have found that under chosen, “in vitro like in vivo” conditions 20 nM SkQ1 effectively reduced cell death in RPE and choroid, protected RPE from disintegration caused by cell phenotypic transformation and withdrawal from the layer, suppressed transmigration of choroidal coat cells. In the ex vivo model we used degenerative processes were more pronounced in the eye cup center where SkQ1 effect was most vivid. All this give us hopes for effectiveness of SkQ1 treatment of retinal central part that is very susceptible to light-induced over-oxidation injury and mostly suffering in many age-related diseases, AMD, in particular.

The new antioxidant 1-benzoyl-6-hydroxy-2,2,4-trimethyl-1,2- dihydroquinoline has a protective effect against carbon tetrachloride-induced hepatic injury in rats

Liver diseases with the central pathogenetic mechanism of oxidative stress are one of the main causes of mortality worldwide.Therefore,dihydroquinoline derivatives,which are precursors of hepatoprotectors and have antioxidant activity,are of interest.We have previously found that some compounds in this class have the ability to normalize redox homeostasis under experimental conditions.Here,we initially analyzed the hepatoprotective potential of the dihydroquinoline derivative 1-benzoyl-6-hydroxy-2,2,4-trimethyl-1,2-dihydroquinoline(BHDQ)for carbon tetrachloride(CCl4)-induced liver injury in rats.Results suggested that BHDQ normalized the alanine aminotransferase,aspartate aminotransferase,and gamma-glutamyl transpeptidase in serum.We also observed an improvement in liver tissue morphology related to BHDQ.Animals with CCl4-induced liver injuries treated with BHDQ had less oxidative stress compared to animals with CCl4-induced liver injury.BHDQ promoted activation changes in superoxide dismutase,catalase,glutathione peroxidase,glutathione reductase,and glutathione transferase on control values in animals with CCl4-induced liver injury.BHDQ also activated gene transcription in Sod1 and Gpx1 via nuclear factor erythroid 2-related factor 2 and forkhead box protein O1 factors.Therefore,the compound of concern has a hepatoprotective effect by inhibiting the development of necrotic processes in the liver tissue,through antioxidation.

Antioxidant Activity Evaluation in a Series of Heterocyclic Compounds Derived from 1,8-Diaminonaphthalene

From (2,3-dihydro-1H-perimidin-2-yl)-phenyl, the substitution of OH group in ortho or para position on the phenyl ring, allows us to synthesize the studied compounds. These three compounds have been characterized by conventional spectroscopic methods (NMR and MS). The interest of this work is to review the antioxidant activity of our compounds. The antioxidant activity screening carried out according to FRAP and DPPH methods revealed significant anti-free radical properties for compounds 1 and 2 even at low concentrations. In contrast to the compound 2, compound 3 for which the OH group is substituted in para position has the lowest activity in both cases. Therefore the para position seems to be the least sensitive position to increase the antioxidant activity of this pharmacophore.

Synthetic Approach for Novel Fluorine Substituted <i>α</i>-Aminophosphonic Acids Containing 1,2,4-Triazin-5-One Moiety as Antioxidant Agents

Novel fluorine substituted α-amino phosphonic acids containing 1,2,4-triazin- 5-one (6a-f) have been obtained from fluoroacylation of 6-(2&prime;-amino-5&prime;-nitrophenyl)-3-thioxo-1,2,4-triazin-5(4H)-one (1) followed by ammonilysis to give the corresponding 3-amino-derivative 3. Condensation of compound 3 with nitro/halogenated aromatic aldehydes yielded the Schiff bases 4. The simple addition of diethyl phosphonate to compound 4 produced the α-amino phosphonates 5. Acidic hydrolysis of compound 5 produced the fluorine substituted α-amino acids derivatives 6. Structures of the new compounds have been established with the help of elemental analysis and spectral measurements. Also, the products evaluated as antioxidants, where the fluorinated α-amino phosphonic acids 6 are more active than the other synthesized systems.

New antioxidant SkQ1 is an effective protector of rat neural retina under conditions of long-term organotypic cultivation

During life human eye is constantly exposed to sunlight and artificial light, the sources of reactive oxygen species (ROS)—the main cause of age-related eye pathology. A novel mitochondria-targeted antioxidant SkQ1 has recently been invented to reduce mitochondrial ROS by cleaning the mitochondria matrix, “the dirtiest place in the cell” in respect of ROS production and accumulation. Earlier we studied SkQ1 effects upon retinal pigment epithelium and choroid in the rat eye posterior cups exposed to long-term 3D organotypic culturing. It was found that under in vitro conditions 20 nM SkQ1 effectively reduced cell death in retinal pigment epithelium and choroid and protected the tissues from disintegration and cell withdrawal. In the present study we used same ex vivo conditions to examine the effect of SkQ1 upon the rat neural retina kept in the content of the posterior eye cup. Eye cups were isolated and cultured in vitro during 7, 14, and 30 days under rotation in the presence and absence of 20 nM SkQ1 in the culture medium. Serial sections of cultivated eye cups were subjected to histology, computer morphometry and immunohistochemistry. Obtained results show that SkQ1 operates as a strong protective agent, preventing neuronal cell death and other degenerative processes in the neural retina. Cell rescue by SkQ1 was more vivid in the central part of the retina than at the periphery. That, in turn, suggests SkQ1 effectiveness in treatment of some age-related eye diseases when central part of the retina, including macula, is most susceptible to degeneration.

Correlation of serum Aβ1-42 content with inflammatory factors and receptors as well as antioxidant enzymes in patients with Parkinson's Disease

Objective: To investigate the correlation of serum β-amyloid 1-42 (Aβ1-42) content with inflammatory factors and receptors as well as antioxidant enzymes in patients with Parkinson's Disease. Methods: A total of 48 patients with Parkinson's disease who were treated in this hospital between December 2014 and October 2017 were selected as Parkinson's disease group, and 50 healthy volunteers who received physical examination in this hospital during the same period were selected as normal control group. The differences in serum contents of Aβ1-42, inflammatory factors and receptors as well as antioxidant enzymes were compared between the two groups, and Pearson test was used to assess the correlation between serum Aβ1-42 content and illness in patients with Parkinson's disease. Results: Serum Aβ1-42 content of Parkinson's disease group was lower than that of normal control group;serum inflammatory factors and receptors IL-2, sIL-2R, IL-6 and sIL-6R contents were higher than those of normal control group;serum antioxidant enzymes SOD, GSH-Px, CAT and TPX contents were lower than those of control group. Pearson test showed that serum Aβ1-42 content of patients with Parkinson's disease was directly correlated with the contents of inflammatory factors and receptors as well as antioxidant enzymes. Conclusions: Serum Aβ1-42 content decreases in patient with Parkinson's disease, and the specific content is directly correlated with the condition of Parkinson's disease, and can be used as an important auxiliary indicator for diagnosis and judgment of Parkinson's disease.

间苯三酚联合益气养阴汤治疗HDP对患者血清HMGB1、TAC、MDA水平及分娩结局影响

目的:分析益气养阴汤联合间苯三酚治疗妊娠期高血压(HDP)对患者血清高迁移率蛋白-1(HMGB1)、总抗氧化能力(TAC)、丙二醛(MDA)水平及分娩结局影响。方法:选取2021年1月-2022年12月本院收治的HDP患者96例,按随机数字表法分为对照组(48例,给予间苯三酚治疗)和联合组(48例,在对照组基础上给予益气养阴汤治疗)。比较两组疗效、血清HMGB1、TAC、MDA水平、脂代谢指标、血压、不良反应及分娩结局。结果:治疗后联合治疗组总有效率(95.8%)高于对照组(77.1%),两组舒张压、收缩压、平均动脉压均低于治疗前且联合组低于对照组,两组血清HMGB1、MDA水平均低于治疗前且联合组(1.85±0.47mg/L、23.56±3.56μmol/L)低于对照组(3.36±0.82 mg/L、32.47±5.16μmol/L),两组TAC水平均升高且联合组(11.26±2.65U/ml)高于对照组(9.24±2.03U/ml),两组甘油三酯、总胆固醇、低密度脂蛋白胆固醇水平均降低且联合组低于对照组,两组高密度脂蛋白胆固醇水平均上调且联合组高于对照组(均P<0.05)。联合组不良妊娠结局总发生率(8.3%)低于对照组(37.5%)(P=0.001),不良反应总发生率(6.3%)与对照组(12.5%)无差异(P=0.484)。结论:益气养阴汤联合间苯三酚治疗HDP,可更好控制患者血压水平,改善氧化应激和脂代谢水平,改善母婴结局,提高临床治疗效果,安全性好。

ABS树脂抗氧剂YHK-1复配条件的选择

抗氧剂YHK - 1主要由酚类和亚磷酸酯类抗氧剂组成 ,能有效抑制聚合物氧化 ,被广泛用于ABS树脂中。由于它是复合型抗氧剂 ,复配条件的选择很重要。通过一系列的实验 ,总结出最佳复配条件为温度 90～ 10 0℃ ,时间 1 5～ 2 5h ,搅拌速度

抗氧剂YHK-1在ABS树脂中的性能评价

抗氧剂YHK-1由酚类抗氧剂、亚磷酸酯类抗氧剂及特殊添加剂组成。将自制抗氧剂YHK-1与进口同类产品进行比较,发现YHK-1抗氧剂能够更有效地抑制聚合物氧化,完全可替代进口产品而应用于ABS树脂中。

1-甲基环丙烯对根用芥菜贮藏品质及抗氧化性的影响

为解决根用芥菜贮藏期失重和发芽的问题,本研究以光头芥为试验材料,对照组不做处理,处理组采用1.0、1.5和2.0μL·L^(-1)1-甲基环丙烯(1-MCP)进行熏蒸,探究不同浓度1-MCP处理对低温贮藏条件下根用芥菜贮藏品质及抗氧化性的影响。结果表明,1-MCP处理可提高根用芥菜的低温贮藏品质并促进其抗氧化作用。与对照相比,不同浓度1-MCP处理有效抑制了根用芥菜的呼吸强度,提高了贮藏品质及抗氧化作用,其中1.0μL·L^(-1)1-MCP对根用芥菜的失重率、发芽率和木质素含量的抑制作用显著高于其他处理组(P<0.05);1-MCP处理保持了较高的硬度和抗坏血酸含量、延缓了根用芥菜细胞破损和丙二醛(MDA)含量积累、抑制了脂氧合酶(LOX)和苯丙氨酸解氨酶(PAL)活性、减缓了过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性下降、提高了超氧化物歧化酶(SOD)和过氧化物酶(POD)活性,其中1.0μL·L^(-1)1-MCP效果最优。本研究结果为根用芥菜的贮藏保鲜提供了理论依据。

GLP-1受体激动剂和DPP-4抑制剂分别联合二甲双胍治疗T2DM的疗效及对血清抗氧化因子、炎症因子的影响

目的对比胰高血糖素样肽-1(GLP-1)受体激动剂和二肽基肽酶-4(DPP-4)抑制剂分别联合二甲双胍治疗2型糖尿病(T2DM)的疗效及对其血清抗氧因子、炎症因子的影响。方法将2020年5月至2022年5月中国中医科学院广安门医院南区收治的120例T2DM患者纳入本次前瞻性研究,采用随机数字表法分成GLP-1联合组(n=40)、DPP-4联合组(n=40)和对照组(n=40)。GLP-1联合组患者治疗方案为二甲双胍治疗+利拉鲁肽,DPP-4联合组患者治疗方案为二甲双胍+沙格列汀,对照组患者仅口服二甲双胍治疗。治疗为期6个月。比较3组患者治疗前后的血糖相关指标[空腹血糖、餐后2 h血糖(2 h PPG)、糖化血红蛋白(HbA1c)]及胰岛素抵抗指数(HOMA-IR)、氧化应激指标[超氧化物歧化酶(SOD)和6-酮-前列环素F1α(6-Keto-PGF1α)]和炎症因子[肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和超敏C反应蛋白(hs-CRP)]表达情况。结果GLP-1联合组治疗后的空腹血糖、2 hPPG、HbA1c和HOMA-IR分别为(5.05±0.69)mmol/L、(7.42±0.98)mmol/L、(6.11±0.70)%和(3.20±0.45),DPP-4联合组分别为(5.58±0.61)mmol/L、(8.09±1.04)mmol/L、(6.65±0.76)%和(3.78±0.50),对照组分别为(6.50±0.75)mmol/L、(10.14±1.22)mmol/L、(7.80±0.81)%和(4.61±0.59),GLP-1联合组与DPP-4联合组治疗后的上述血糖相关指标及HOMA-IR均较对照组显著下降,且GLP-1联合组显著低于DPP-4联合组,差异均有统计学意义(P<0.05)。GLP-1联合组治疗后的SOD和6-Keto-PGF1α分别为(95.09±9.97)U/mL、(76.19±6.80)ng/L,DPP-4联合组分别为(85.17±10.18)U/mL、(67.32±6.39)ng/L,对照组分别为(76.89±9.06)U/mL、(60.46±6.02)ng/L,GLP-1联合组与DPP-4联合组治疗后的血清SOD、6-Keto-PGF1α均较对照组显著上升,且GLP-1联合组血清SOD、6-Keto-PGF1α均较DPP-4联合组显著上升,差异均有统计学意义(P<0.05)。GLP-1联合组TNF-α、IL-6和hs-CRP分别为(7.05±1.16)ng/L、(5.01±1.35)pg/mL、(4.04±0.51)mg/L,DPP-4联合组为(7.93±1.29)ng/L、(5.97±1.40)pg/mL、(4.99±0.59)mg/L,对照组为(10.34±1.58)ng/L、(7.58±1.49)pg/mL、(5.94±0.64)mg/L,GLP-1联合组与DPP-4联合组治疗后的上述血清炎症因子均较对照组显著下降,且GLP-1联合组显著低于DPP-4联合组,差异均有统计学意义(P<0.05)。结论GLP-1受体激动剂联合二甲双胍治疗T2DM的降糖效果优于DPP-4受体抑制剂联合二甲双胍,在改善胰岛素抵抗和氧化应激,降低炎症反应方面同样优于DPP-4受体抑制剂联合二甲双胍治疗。

急性脑梗死患者外周血Keap1⁃Nrf2/ARE信号通路变化与预后的相关性分析

目的分析急性脑梗死(ACI)患者外周血Kelch样环氧氯丙烷相关蛋白⁃1(Keap1)⁃核因子E2相关因子2(Nrf2)/抗氧化反应元件(ARE)信号通路变化与预后的相关性。方法选取2020年6月—2022年3月安徽医科大学附属巢湖医院神经内科诊治ACI患者110例作为研究对象,根据溶栓后3个月的改良Rankin量表(mRs)评分分为近期预后良好组与近期预后不良组,随访1年ACI是否复发分为远期预后良好组与远期预后不良组,比较不同近远期预后组溶栓前、溶栓后7 d、溶栓后3个月外周血Keap1、醌氧化还原酶1(NQO1)、Nrf2、ARE蛋白相对表达量、神经功能缺损(NIHSS)评分,分析各蛋白与NIHSS评分的相关性,受试者工作特征曲线(ROC)评价各蛋白对预后的预测价值。结果110例ACI患者,近期预后良好患者78例,近期预后不良患者32例;随访1年,失访6例,远期预后良好患者83例,远期预后不良患者21例。近远期预后不良组溶栓后7 d、3个月Keap1蛋白相对表达量、NIHSS评分高于近远期预后良好组,NQO1、Nrf2、ARE蛋白相对表达量低于近远期预后良好组(P均<0.001);溶栓后7 d、3个月Keap1蛋白相对表达量与NIHSS评分呈正相关(r=0.684、0.702,P均<0.001),NQO1、Nrf2、ARE蛋白相对表达量与NIHSS评分呈负相关(7 d:r=-0.615、-0.628、-0.609,3个月:r=-0.631、-0.657、-0.629,P均<0.001);溶栓后7 d的Keap1、NQO1、Nrf2、ARE蛋白相对表达量及四者联合预测近期预后的AUC为0.803、0.717、0.800、0.765、0.907,四者联合优于各蛋白单独预测(Z/P=2.105/0.026、2.413/0.004、2.113/0.021、2.205/0.016);溶栓后3个月的Keap1、NQO1、Nrf2、ARE蛋白相对表达量及四者联合预测远期预后不良的AUC为0.766、0.765、0.812、0.767、0.927,四者联合优于各蛋白单独预测(Z/P=2.238/0.010、2.241/0.008、2.115/0.019、2.231/0.012);溶栓后3个月的外周血Keap1⁃Nrf2/ARE蛋白预测远期预后的AUC与溶栓后7 d的外周血Keap1⁃Nrf2/ARE蛋白预测近期预后的AUC比较,差异无统计学意义(Z/P=0.102/0.563)。结论ACI患者外周血Keap1⁃Nrf2/ARE信号通路变化与神经功能缺损程度、近远期预后显著相关,可从相关蛋白含量变化的角度有效预测近远期预后情况。

基于Keap1/Nrf2/HO-1信号通路和氧化应激效应研究白肉灵芝多糖对小鼠运动性疲劳的保护作用

目的:研究白肉灵芝多糖对高强度训练小鼠的抗疲劳作用机制。方法:小鼠随机分为空白对照组、模型对照组和白肉灵芝多糖低、中、高3个处理组,通过跑台训练建立小鼠运动性疲劳模型,处理组按照多糖浓度10、50、100 mg/mL分别连续给予多糖灌胃,其他组灌胃相同剂量的无菌生理盐水,连续训练4周后进行小鼠爬杆实验,记录小鼠爬杆时间,并测定小鼠体内睾酮(TTS)、皮质酮(CTC)、血尿素氮(BUN)、血乳酸(BLA)、肝糖原(HG)、肌糖原(MG)、丙二醛(MDA)含量和过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物歧化酶(GSH-Px)等活力,同时Western blot检测小鼠骨骼肌中Keap-1、Nrf2、HO-1的蛋白表达量。结果:低、中、高多糖处理组小鼠爬杆时间显著提高(P <0.05),同时小鼠血清中TTS含量逐渐升高,CTC含量则逐渐下降,HG和MG的含量逐渐升高;另外多糖低、中、高剂量组小鼠血清和肝组织中MDA含量均显著降低(P <0.05),CAT、SOD、GSH-Px酶活性均显著提高(P <0.05)。与空白对照组相比,模型组中Keap1的蛋白表达水平显著升高(P <0.05),并下调了Nrf2和HO-1的蛋白表达,与模型组相比白肉灵芝多糖低、中、高多糖处理组的Keap1蛋白表达水平依次下降,Nrf2和HO-1的蛋白表达水平则依次升高。结论:白肉灵芝多糖对小鼠运动性疲劳的保护作用可能是与提高小鼠骨骼肌细胞抗氧化作用及影响Keap1/Nrf2/HO-1信号通路有关。

补阳还五汤对一氧化碳中毒迟发性脑病大鼠氧化损伤及Nrf2/HO-1信号通路的影响

目的:探讨补阳还五汤对一氧化碳中毒迟发性脑病(delayed encephalophathy after acute carbon monoxide poisoning,DEACMP)大鼠氧化损伤的脑组织保护作用及其对核转录因子2(nuclear transcription factor 2,Nrf2)/血红素氧合酶1(heme oxygenose 1,HO-1)信号通路的影响。方法:采用改良式间隔腹腔注射法制作DEACMP大鼠模型;应用血气分析仪测定碳氧血红蛋白(carboxyhemaglobin,COHb)浓度;应用Morris水迷宫试验观察大鼠行为学表现;将造模成功的大鼠随机分为模型组(n=30)和补阳还五汤(BHD)高、中、低剂量组(n=30),以注射等量空气作为正常组(AC组);HE染色方法检测大鼠海马区病理变化;流式细胞术检测大鼠海马组织细胞凋亡情况;生化实验方法检测大鼠血液中丙二醛(malonaldehyde,MDA)、8-羟基脱氧鸟苷(8-hydroxy-2 deoxyguanosine,8-OHdG)水平;Western blot和qPCR方法检测大鼠海马组织中血红素氧合酶1(heme oxygenose 1,HO-1)、核因子红细胞系相关因子2(nuclear transcription factor 2,Nrf2)、Kelch样环氧氯丙烷相关蛋白1(Keap1)、磷酸化Nrf2(p-Nrf2)蛋白及其mRNA表达;蛋白质免疫共沉淀(Co-Immunoprecipitation,CoIP)方法检测Nrf2与Keap1的相互作用。结果:补阳还五汤可改善DEACMP模型大鼠海马组织损伤,减少炎症细胞浸润和组织水肿,降低海马组织细胞凋亡;提高海马组织HO-1、Nrf2、p-Nrf2蛋白及其mRNA表达,降低Keap1蛋白及其mRNA表达水平;提高海马组织细胞核中Nrf2蛋白与Keap1蛋白的相互作用量;降低大鼠血液中MDA和8-OHdG含量。结论:补阳还五汤可激活大鼠海马组织中Nrf2/HO-1信号通路,进而保护海马组织免遭急性一氧化碳中毒所致的氧化应激损伤,能抑制大鼠海马区神经元的持续凋亡,进而预防DEACMP。