Correlative Expression of Glutathion S-Transferase-π and Multidrug Resistance Associated Protein in Bladder Transitional Cell Carcinoma

In order to elucidate the mechanisms of multidrug resistance (MDR) in bladder cancer, the expression of glutathione S-transferase-π (GST-π) and multidrug resistance associated protein (MRP) in tissue samples resected from 44 patients and 6 normal bladder mucosa as control was de- tected by using immunohistochemical method, and the results were analyzed by computer-assisted im- age analyzing system (IAS) to achieve semi-quantitative data. In addition, correlation between the expression of both factors was studied. The results showed that the positive expression rate of GST- π and MRP in bladder cancer was 72. 7 % (32/44) and 68. 2 % (30/44) respectively, significantly higher than those in normal bladder mucosa, being 16. 7% and 33. 3% respectively. The rate of GST-πpositive staining was increased correspondingly with tumor grade and stage elevated, being higher in recurrent tumors treated by chemotherapy, but not significantly (P>0. 05). There was no significant differences between the expression of MRP and tumors' behaviors and clinical characters. However, the results demonstrated that the correlation between the expression of both resistant fac- tors was very evident (r=0. 695, P<0. 0025). It was suggested that the activation of GST-π and MRP might occur during malignant transformation of normal mucosa, but tumors' differentiation and progression could not be the unique factors that influenced both overexpression. Chemotherapy might be another important reason. The correlation of both indicated that there was a common mech- anism regulating their expression probably, which made them play a pivotal role in chemotherapy drug resistance of bladder cancers.

The effect of glutathione on glucosinolate biosynthesis through the sulfur assimilation pathway in pakchoi associated with the growth conditions

Glucosinolates(GSLs) are a group of nitrogen-and sulfur-containing secondary metabolites, synthesized primarily in members of the Brassicaceae family, that play an important role in food flavor, plant antimicrobial activity, resistance to insect attack, stress tolerance, and human anti-cancer effects. As a sulfur-containing compound, glutathione has a strong connection with GSLs biosynthesis as a sulfur donor or redox system, and exists in reduced(glutathione;GSH) and oxidized(glutathione disulfide;GSSG) forms. However, the mechanism of GSH regulating GSLs biosynthesis remainds unclear. Hence, the exogenous therapy to pakchoi under normal growth condition and sulfur deficiency condition were conducted in this work to explore the relevant mechanism. The results showed that exogenous application of buthionine sulfoximine, an inhibitor of GSH synthesis, decreased the transcript levels of GSLs synthesis-related genes and transcription factors, as well as sulfur assimilation-related genes under the normal growth condition. Application of exogenous GSH inhibited the expression of GSLs synthesis-and sulfur assimilation-related genes under the normal condition, while the GSLs biosynthesis and the sulfur assimilation pathway were activated by exogenous application of GSH when the content of GSH in vivo of plants decreased owing to sulfur deficiency. Moreover,exogenous application of GSSG increased the transcript levels of GSLs synthesis-and sulfur assimilation-related genes under the normal growth condition and under sulfur deficiency. The present work provides new insights into the molecular mechanisms of GSLs biosynthesis underlying glutathione regulation.

Silent information regulator sirtuin 1 ameliorates acute liver failure via the p53/glutathione peroxidase 4/gasdermin D axis

BACKGROUND Acute liver failure(ALF)has a high mortality with widespread hepatocyte death involving ferroptosis and pyroptosis.The silent information regulator sirtuin 1(SIRT1)-mediated deacetylation affects multiple biological processes,including cellular senescence,apoptosis,sugar and lipid metabolism,oxidative stress,and inflammation.AIM To investigate the association between ferroptosis and pyroptosis and the upstream regulatory mechanisms.METHODS This study included 30 patients with ALF and 30 healthy individuals who underwent serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)testing.C57BL/6 mice were also intraperitoneally pretreated with SIRT1,p53,or glutathione peroxidase 4(GPX4)inducers and inhibitors and injected with lipopolysaccharide(LPS)/D-galactosamine(D-GalN)to induce ALF.Gasdermin D(GSDMD)^(-/-)mice were used as an experimental group.Histological changes in liver tissue were monitored by hematoxylin and eosin staining.ALT,AST,glutathione,reactive oxygen species,and iron levels were measured using commercial kits.Ferroptosis-and pyroptosis-related protein and mRNA expression was detected by western blot and quantitative real-time polymerase chain reaction.SIRT1,p53,and GSDMD were assessed by immunofluorescence analysis.RESULTS Serum AST and ALT levels were elevated in patients with ALF.SIRT1,solute carrier family 7a member 11(SLC7A11),and GPX4 protein expression was decreased and acetylated p5,p53,GSDMD,and acyl-CoA synthetase long-chain family member 4(ACSL4)protein levels were elevated in human ALF liver tissue.In the p53 and ferroptosis inhibitor-treated and GSDMD^(-/-)groups,serum interleukin(IL)-1β,tumour necrosis factor alpha,IL-6,IL-2 and C-C motif ligand 2 levels were decreased and hepatic impairment was mitigated.In mice with GSDMD knockout,p53 was reduced,GPX4 was increased,and ferroptotic events(depletion of SLC7A11,elevation of ACSL4,and iron accumulation)were detected.In vitro,knockdown of p53 and overexpression of GPX4 reduced AST and ALT levels,the cytostatic rate,and GSDMD expression,restoring SLC7A11 depletion.Moreover,SIRT1 agonist and overexpression of SIRT1 alleviated acute liver injury and decreased iron deposition compared with results in the model group,accompanied by reduced p53,GSDMD,and ACSL4,and increased SLC7A11 and GPX4.Inactivation of SIRT1 exacerbated ferroptotic and pyroptotic cell death and aggravated liver injury in LPS/D-GalNinduced in vitro and in vivo models.CONCLUSION SIRT1 activation attenuates LPS/D-GalN-induced ferroptosis and pyroptosis by inhibiting the p53/GPX4/GSDMD signaling pathway in ALF.

Novel insights into conjugation of antitumor-active unsymmetrical bisacridine C-2028 with glutathione:Characteristics of non-enzymatic and glutathione S-transferase-mediated reactions

Unsymmetrical bisacridines(UAs) are a novel potent class of antitumor-active therapeutics.A significant route of phase II drug metabolism is conjugation with glutathione(GSH),which can be non-enzymatic and/or catalyzed by GSH-dependent enzymes.The aim of this work was to investigate the GSHmediated metabolic pathway of a representative UA,C-2028.GSH-supplemented incubations of C-2028 with rat,but not with human,liver cytosol led to the formation of a single GSH-related metabolite.Interestingly,it was also revealed with rat liver microsomes.Its formation was NADPH-independent and was not inhibited by co-incubation with the cytochrome P450(CYP450) inhibitor 1-aminobenzotriazole.Therefore,the direct conjugation pathway occurred without the prior CYP450-catalyzed bioactivation of the substrate.In turn,incubations of C-2028 and GSH with human recombinant glutathione S-transferase(GST) P1-1 or with heat-/ethacrynic acid-inactivated liver cytosolic enzymes resulted in the presence or lack of GSH conjugated form,respectively.These findings proved the necessary participation of GST in the initial activation of the GSH thiol group to enable a nucleophilic attack on the substrate molecule.Another C-2028-GSH S-conjugate was also formed during non-enzymatic reaction.Both GSH S-conjugates were characterized by combined liquid chromatography/tandem mass spectrometry.Mechanisms for their formation were proposed.The ability of C-2028 to GST-mediated and/or direct GSH conjugation is suspected to be clinically important.This may affect the patient’s drug clearance due to GST activity,loss of GSH,or the interactions with GSH-conjugated drugs.Moreover,GST-mediated depletion of cellular GSH may increase tumor cell exposure to reactive products of UA metabolic transformations.

Oxidative stress regulated heme-oxygenase-1 and glutathione S-transferase-m1 gene expression changes in cell lines exposed to melanins

To investigate the effects of oxidative stress on substantia nigra neuronal degeneration and death in patients with Parkinson＇s disease, we treated neuroblastoma cells （SK-N-SH） and glioma cells with Fenton＇s reagent, iron chelating agent, neuromelanin and dopamine melanin. We investigated the changes in expression of nine oxidative stress-related genes and proteins. The levels of mRNAs for heme-oxygenase-1 and glutathione S-transferase-ml were significantly reduced in SK-N-SH cells exposed to oxidative stress, and increased in glial cells treated with deferoxamine. These results revealed that SK-N-SH neurons react sensitively to oxidative stress, which implies different outcomes between these two types of cells in the substantia nigra. Moreover, the influences of neuromelanin and dopamine melanin on cell function are varied, and dopamine melanin is not a good model for neuromelanin.

Immunohistochemical localization of glutathione S-transferase-pi in human colorectal polyps

AIM:To investigate the distribution of the placental form of glutathione-S-transferase (GST) in colon polyps in order to evaluate the role of GST-pi in these tissues. METHODS: Sixteen polyp tissues removed at colonoscopy were examined. Tissues were investigated histologically and ultrastructurally. GST-pi expression was also analysed immunohistochemically, using peroxidase anti-peroxidase (PAP) method and immunogold labelling method, for light and electron microscope respectively. RESULTS: All polyp tissues examined were adenoma of low, mild and high-grade dysplasia as shown in the histopathological reports. Nevertheless, the examination of the above specimens with electron microscope revealed that 3 of 9 adenoma of mild dysplasia had ultrastuctural features similar to high-grade dysplasia adenoma. GST-pi was variably expressed in adenoma, with the lowest relative levels occurring in low-gradeadenoma and the highest levels found in high-grade adenoma. GST-pi was located mainly in undifferentiated epithelial cells. GST-pi positive particles were found in the cytoplasm and especially in the nucleus adjacent to the nuclear membrane of these cells. CONCLUSION:The overexpression of GST-pi in mildgrade adenomas with significant subcellular changes and in the majority of high-grade dysplasia adenoma suggests that this might be related to the carcinogenetic proceeding. Immunohistochemical localization of GST-pi in combination with ultrastructural changes indicate that GST-pi might be a sensitive agent for the detection of preneoplastic transformations in adenoma.

N-acetylcysteine and reduced glutathione reverse flupirtine-induced liver injury and pro⁃duce other beneficial effects in combination with flupirtine

OBJECTIVE To assess whether N-acetylcysteine(NAC)and reduced glutathione(GSH)are effective in reversing flupirtine-induced hepatotoxicity and whether they have other beneficial effects when combined with flupirtine.METHODS The analgesic effects of NAC and flupirtine were first evaluated in carrageenaninduced inflammatory pain and paclitaxel-induced neuropathic pain.The combination subthreshold⁃ing approach was then used to determine whether the combination of NAC and flupirtine produced synergistic analgesic effects.Hepatotoxicity markers and histopathological examination of the liver were used to assess the efficacy of NAC and GSH in reversing flupirtine-induced hepato⁃toxicity.Finally,the effect of GSH on the safe range of flupirtine was assessed in an acute tox⁃icity assay.RESULTS Flupirtine and NAC pro⁃duced dose-dependent antiallodynic effects evoked by carrageenan and paclitaxel in mice.In the above model,the combination of NAC and flupirtine produced an unexpected synergistic analgesic effect.There were no significant differ⁃ences observed in the hepatotoxicity markers and liver histopathology between the experimen⁃tal group and the control group under NAC and GSH treatment.Finally,GSH(200 mg·kg^(-1))expanded the therapeutic index of flupirtine by 1.77 times.CONCLUSION NAC and GSH are effective in preventing liver damage caused by long-term flupirtine use,which provides a solu⁃tion for the safe and effective treatment of chronic pain with flupirtine.In addition,the other benefi⁃cial effects of NAC and GSH when combined with flupirtine may provide the basis for the devel⁃opment of a new therapy with minimal sideeffects and good efficacy.

Molecular identification and biochemical characteristics of a delta class glutathione S-transferase gene(FcδGST)from Chinese shrimp Fenneropenaeus chinensis

Glutathione S-transferases(GSTs)are a superfamily of multifunction enzymes involved in the regulation of redox homeostasis and innate immune responses against various pathogenic infections in marine invertebrates.In the present study,a delta class GST gene(designated as FcδGST)was cloned from Fenneropenaeus chinensis using rapid amplification of c DNA ends(RACE)technology.The complete cDNA sequence of FcδGST was 780 bp in length,which includes a 27-bp 5′non-coding region(UTR),a 117-bp 3′UTR,a 636-bp open reading frame(ORF),and a polyadenylate signal site(AATAAA)presented at the upstream of poly A tail.The FcδGST gene encoded 211 amino acids peptide,including a GST_N domain and a GST_C domain,and exhibited high similarity with previously reported delta GSTs.The predicted molecular mass of FcδGST protein was 23.39 kDa,and its theoretical isoelectric point(pI)was 5.34.The FcδGST mRNA transcripts were ubiquitously expressed in all the tested tissues,with the highest expression level in hemocytes and hepatopancreas.During the stimulation of Vibrio anguillarum or white spot syndrome virus(WSSV),the m RNA expression of FcδGST in hemocytes and hepatopancreas revealed significant up-regulation.The purified recombinant FcδGST protein(designated as rFcδGST)exhibited specific catalytic activity against 1-chloro-2,4-dinitrobenzene(CDNB)substrate with relatively low stable enzymatic activities.These results indicated that FcδGST was a fragile but typical novel delta class GST member and potentially involved in the innate immune responses of F.chinensis.

Evaluation of Glutathione Peroxidase Enzymatic Activity in Seminal Plasma of Patients Treated at the Institute Pasteur in Cote d’Ivoire

Glutathione peroxidase (GPx) is an antioxidant that plays an important role in the maintenance of male fertility. The aim of this study was to compare the profile of enzymatic activity of glutathione peroxidase in the seminal plasma of normozoosperm and those of pathological sperm. Thus, the activity of glutathione peroxidase was determined in the seminal plasma of 20 normozoosperms, 9 azoosperms and 31 oligoasthenoteratozoosperms. It was 37.58 ± 3.14 U/L in normozoosperms, 39.39 ± 2.27 U/L in oligoasthenoteratozoosperms, and 29.77 ± 2.62 U/L in azoosperms. The mean GPx enzyme activity of normozoosperms did not differ significantly from that of oligoasthenoteratozoosperms and azoosperms. In contrast, comparison of enzyme activity between abnormal sperms gave a significant difference. This study showed that glutathione peroxidase enzymatic activity is not related to sperm quality.

Glutathione S-Transferase(GST)Identified from Giant Kelp Macrocystis pyrifera Increases the Copper Tolerance of Synechococcus elongatus PCC 7942

The glutathione S-transferases gene family plays an important regulatory role in growth and development,and responses to environmental change.In this study,six complete GST genes(Mp GST1,Mp GST2,Mp GST3,MpGST4,Mp GST5,and Mp GST6)were cloned from the gametophytes of brown alga Macrocystis pyrifera.Subsequent bioinformatics analysis showed that these six genes encoded proteins with 202,216,288,201,205,and 201 aa,respectively.Moreover,Mp GST3 differs from the other GST genes.Phylogenetic analysis suggested that MpGST3 belongs to the Ure2p type GST.Domain analysis suggested that the other GSTs from M.pyrifera belong to the soluble GST family and form an independent branch with the GSTs found in the other macroalgae,suggesting that a new GST type was formed during macroalgal evolution.GST genes were upregulated in M.pyrifera when 2.5 mg L^(-1)Cu ions were added to the medium.Six GST genes were integrated into the genome of Synechococcus elongatus PCC 7942,and their functions were verified by measuring light absorbance,photosynthetic pigment content,and photosynthetic parameters of the transformed strains under 0.3 mg L^(-1)Cu ion stress.The results showed much higher levels of various parameters in the transformed strains than in the wild strain.The transformed strains(with the MpGST genes)showed significantly enhanced resistance to Cu ion stress,while the wild strain almost died.The results of this study lay a theoretical foundation for further research on the Cu ion stress resistance function of GSTs in M.pyrifera.

DMMIC derivatization-assisted liquid chromatography-mass spectrometry method for metabolite profiling of the glutathione anabolic pathway in esophageal cancer tissues and cells

In this work,a new pyrylium derivatization-assisted liquid chromatography-mass spectrometry(LC-MS)method was developed for metabolite profiling of the glutathione anabolic pathway(GAP)in cancer tissues and cells.The pyrylium salt of 6,7-dimethoxy-3-methyl isochromenylium tetrafluoroborate(DMMIC)was used to label the amino group of metabolites,and a reductant of dithiothreitol(DTT)was employed to stabilize the thiol group.By combining DMMIC derivatization with LC-MS,it was feasible to quantify the 13 main metabolites on the GAP in complex biological samples,which had good linearity(R^(2)=0.99810.9999),precision(interday precision of 1.6%e19.0%and intraday precision of 1.4%e19.8%)and accuracy(83.4%-115.7%).Moreover,the recovery assessments in tissues(82.5%e107.3%)and in cells(98.1%e118.9%)with GSH-^(13)C2,^(15)N,and Cys-^(15)N demonstrated the reliability of the method in detecting tissues and cells.Following a methodological evaluation,the method was applied successfully to investigate difference in the GAP between the carcinoma and para-carcinoma tissues of esophageal squamous cell carcinoma(ESCC)and the effect of p-hydroxycinnamaldehyde(CMSP)on the GAP in KYSE150 esophageal cancer cells.The results demonstrate that the developed method provides a promising new tool to elucidate the roles of GAP in physiological and pathological processes,which can contribute to research on drugs and diseases.

Electronic Aspects of the Synergistic Antioxidant Interaction of Various Pairs “Phenolic Food Acid and Glutathione” in Their Reactions with the Stable Radical Cation ABTS+·

In the present work, for the first time, the main details of the electronic mechanism of the synergistic antioxidant interaction between different pairs: phenolic food acid and glutathione and the stable radical cation ABTS+· were revealed on the basis of a rigorous analysis of the DFT calculated data. It was shown that among all the studied food acids, only caffeic acid exhibits a clear-cut significant synergistic effect with glutathione. It established the electronic and structural factors underlying the mechanism of the synergistic interaction of the mixture caffeic acid and glutathione in its reaction with ABTS+·. The main causes of this considered synergistic effect are, firstly, the presence of the 3-OH and 4-OH hydroxyl groups in the structure of caffeic acid, secondly, the greater stability of its anion which contains the deprotonated 4-OH hydroxyl group. All other phenolic food acids under study do not possess the given structural particularity and therefore do not show such synergistic effects with glutathione.

Evaluation of combined detection of nuclear factor erythroid 2-related factor 2 and glutathione peroxidase 4 in primary hepatic carcinoma and preliminary exploration of pathogenesis

This study aims to analyze the clinical significance and mechanism of nuclear factor erythroid 2-related factor 2(NRF2)and glutathione peroxidase 4(GPX4)in primary hepatic carcinoma(PHC).Methods:The expression of NRF2 and GPX4 in peripheral blood of patients with PHC was determined to analyze the diagnostic value of the two combined for PHC.The prognostic significance of NRF2 and GPX4 was evaluated by 3-year followup.Human liver epithelial cells THLE-2 and human hepatocellular carcinoma cells HepG2 were purchased,and the expression of NRF2 and GPX4 in the cells was determined.NRF2 and GPX4 aberrant expression vectors were constructed and transfected into HepG2,and changes in cell proliferation and invasion capabilities were observed.Results:The expression of NRF2 and GPX4 in patients with PHC was higher than that in patients with LC or VH(p<0.05),and the two indicators combined was excellent in diagnosing PHC.Moreover,patients with high expression of NRF2 and GPX4 had a higher risk of death(p<0.05).In in vitro experiments,both NRF2 and GPX4 expression was elevated in HepG2(p<0.05).HepG2 activity was enhanced by increasing the expression of the two,vice versa(p<0.05).Conclusion:NRF2 and GPX4 combined is excellent in diagnosing PHC,and promotes the malignant development of PHC.

Glutathione peroxidase mimicry of diphenyl diselenide: Plausible contribution of proteins’ thiols

Organoseleniums are a class of compounds attracting attention across the globe owing to their Glutathione peroxidase(GPx)mimicry,which confers on them a strong antioxidant activity.Diphenyl diselenide(DPDS)is an Organoselenium whose GPx mimetic property has been suggested to rely on the oxidation of non-protein or protein thiols critical to the activities of some sulfhydryl enzymes.This study,therefore investigated the GPx mimic/antioxidant property of DPDS as well as the role of thiols of two key sulfhydryl enzymes,cerebral Na^(+)/K^(+)-ATPase(sodium pump)and hepatic delta-aminolevulinic acid dehydratase(δ-ALAD)in the GPx mimicry of DPDS.Albino Wistar rats were euthanized,and the liver and brain were removed and used to assay for the effect of DPDS on lipid peroxidation induced by two prooxidants[Fe2^(+)(10μM)and H2O2,(1 mM)]as well as the activities of the sulfhydryl enzymes.The results revealed that DPDS profoundly(P<0.05)counteracted Fe2^(+)and H2O2-induced lipid peroxidation in the rats’hepatic and cerebral tissues.Furthermore,the results of assay systems for lipid peroxidation and sodium pump revealed that DPDS inhibited Na^(+)/K^(+)-ATPase and lipid peroxidation in the brain tissue homogenates in the same reaction system.A similar result was obtained in the assay system for lipid peroxidation and hepaticδ-ALAD as DPDS simultaneously inhibited the enzyme’s activity and lipid peroxidation.This suggests that the GPx mimetic property of DPDS may be linked to the enzymes’loss of activity,which further validates the suggestions that the enzymes’inhibition,as well as the antioxidant action of DPDS,rely on the oxidation of critical thiols of the enzymes.However,the GPx mimicry of DPDS should be investigated in the presence of thiol-blocking or oxidizing agents in biological systems in order to further ascertain the role of protein thiols.

铁死亡诱导剂RAS合成致死分子3抑制病理性瘢痕成纤维细胞的纤维化

背景:病理性瘢痕主要表现为异常的细胞外基质积累和过度的成纤维细胞增殖,成纤维细胞过度增殖就会产生大量以胶原纤维为主的细胞外基质。因此深入探讨成纤维细胞纤维化在病理性瘢痕形成中的作用,将为揭示病理性瘢痕的机制和生物学治疗提供新思路。目的:探讨铁死亡诱导剂RAS合成致死分子3(RAS-selective lethal small molecule 3,RSL3)对人病理性瘢痕成纤维细胞纤维化的影响。方法:收集10例宁夏医科大学总医院烧伤整形美容科提供的病理性瘢痕组织和同一个体正常皮肤组织,提取人病理性瘢痕成纤维细胞和人正常皮肤成纤维细胞用于后续实验;苏木精-伊红染色观察病理性瘢痕组织和正常皮肤组织的形态;倒置显微镜观察病理性瘢痕成纤维细胞和正常皮肤成纤维细胞的外观形态;免疫荧光实验验证所提取的细胞是否为成纤维细胞;用不同浓度的RSL3(1,3,5,7,9,11,13μmol/L)干预细胞,CCK-8法检测RSL3作用于成纤维细胞的半数抑制浓度(IC_(50));设置对照组(不做处理)和RSL3干预组(用7μmol/L的RSL3干预细胞24 h),qRT-PCR和Western blot检测谷胱甘肽过氧化物酶4、Ⅰ型胶原蛋白、Ⅲ型胶原蛋白和α-平滑肌肌动蛋白的mRNA和蛋白的表达;检测细胞丙二醛浓度;划痕试验检测细胞划痕后24 h剩余划痕面积,并计算剩余划痕面积百分比。结果与结论:①与正常皮肤组相比,病理性瘢痕组的谷胱甘肽过氧化物酶4高表达(mRNA:t=3.252,P<0.01;蛋白:t=5.075,P<0.01);②与正常皮肤成纤维细胞组相比,病理性瘢痕成纤维细胞组的谷胱甘肽过氧化物酶4高表达(mRNA:t=10.32,P<0.01;蛋白:t=26.22,P<0.01);③与对照组相比,RSL3干预组谷胱甘肽过氧化物酶4表达减少(mRNA:t=2.798,P<0.05;蛋白:t=4.643,P<0.01),丙二醛浓度上升(t=2.917,P<0.05),Ⅰ型胶原蛋白(mRNA:t=15.84,P<0.01;蛋白:t=4.610,P<0.01)、Ⅲ型胶原蛋白(mRNA:t=28.86,P<0.01;蛋白:t=7.713,P<0.01)和α-平滑肌肌动蛋白(mRNA:t=2.671,P<0.05;蛋白:t=7.417,P<0.01)的表达减少,迁移能力减弱(t=14.06,P<0.01);④提示RSL3通过抑制谷胱甘肽过氧化物酶4的表达,进而抑制病理性瘢痕成纤维细胞的纤维化和迁移能力。

偏头痛脑代谢物改变及与认知情绪关系的MRS研究

目的探讨利用MRS技术分析偏头痛患者大脑中的神经递质变化及其与认知、情绪改变的关系。方法选取31例偏头痛(MA)患者为MA组,28例健康受试者作为对照组。使用MRS分析前扣带回皮层(ACC)、小脑齿状核(DN)及左侧岛叶(LINS)的γ氨基丁酸(GABA)、谷氨酸(Glu)和谷胱甘肽(GSH)水平的变化,并观察其与认知、情绪及偏头痛特征的相关性。结果与对照组比较,MA组表现出认知损害和焦虑症状;ACC中GSH水平增高(P=0.010),DN中GABA(P=0.043)和LINS中GABA(P=0.029)水平降低;DN中GABA与连线试验呈负相关(r=-0.542,P=0.002),LINS中GABA与MSQ(r=-0.441,P=0.017)、PHQ-9(r=-0.400,P=0.031)呈负相关。结论本文为偏头痛患者认知、情绪变化的分析提供了新的视角。

铁死亡在不同细菌所致小鼠血流感染模型中的变化规律及生物学意义

背景:发现血流感染新的疾病诊断标志物、治疗疾病及减轻脏器损伤的分子靶点具有重要意义。铁死亡是新发现的一种细胞死亡形式,脓毒症动物模型中铁死亡的过度激活与炎症反应激活以及肝脏、心脏、肾脏等重要脏器的损伤有关,但铁死亡与血流感染的关系尚不十分清楚。目的:探讨铁死亡在不同细菌所致小鼠血流感染模型中的变化规律及生物学意义。方法:建立革兰阴性菌大肠埃希菌、肺炎克雷伯菌及革兰阳性菌金黄色葡萄球菌、粪肠球菌血流感染的SPF级ICR雄性小鼠模型,每组各42只。建模后0.5,1,3,6,12,24,48 h时检测肝脏、心肌、肾脏中铁死亡标志基因转铁蛋白受体1、谷胱甘肽过氧化物酶4 mRNA表达水平。另选用18只SPF级ICR雄性小鼠,随机分为二甲基亚砜(DMSO)对照组、DMSO+肺炎克雷伯菌组、铁死亡抑制剂Ferrostatin-1+肺炎克雷伯菌组,每组6只;后两组采用尾静脉注射肺炎克雷伯菌悬液的方式建立肺炎克雷伯菌血流感染模型,分别在血流感染建模前1 h给予5 mg/kg的Ferrostatin-1及等剂量DMSO腹腔注射;建模后6 h小鼠检测血清中丙氨酸氨基转移酶、天冬氨酸氨基转移酶、血肌酐、血尿素氮、磷酸肌酸激酶同工酶、乳酸脱氢酶以及各组织中铁死亡标志基因的mRNA表达水平。结果与结论:①血流感染建模后,不同细菌血流感染小鼠肝脏、心肌、肾脏中转铁蛋白受体1 mRNA表达水平先升高后降低,谷胱甘肽过氧化物酶4 mRNA表达水平先降低后升高且均在建模后6 h达到峰值;②革兰阴性菌血流感染小鼠中转铁蛋白受体1和谷胱甘肽过氧化物酶4 mRNA表达的变化较革兰阳性菌血流感染小鼠更为显著,其中以肺炎克雷伯菌血流感染小鼠中转铁蛋白受体1和谷胱甘肽过氧化物酶4 mRNA表达的变化最显著;③肺炎克雷伯菌血流感染建模后6 h时,小鼠的丙氨酸氨基转移酶、天冬氨酸氨基转移酶、血肌酐、血尿素氮、磷酸肌酸激酶同工酶、乳酸脱氢酶水平均明显升高;④建模前给予铁死亡抑制剂Ferrostatin-1干预可显著降低丙氨酸氨基转移酶、天冬氨酸氨基转移酶、血肌酐、血尿素氮、磷酸肌酸激酶同工酶、乳酸脱氢酶表达水平;⑤以上结果提示不同细菌致血流感染小鼠中铁死亡明显激活且革兰阴性菌血流感染小鼠的铁死亡激活更为显著;抑制铁死亡可明显减轻肺炎克雷伯菌血流感染小鼠的肝脏、心肌、肾脏损伤。

谷胱甘肽联合西地那非治疗勃起功能障碍患者临床疗效及对血管内皮功能的影响

目的探究谷胱甘肽联合西地那非治疗勃起功能障碍(ED)患者临床疗效及对血管内皮功能、炎性因子、勃起功能的影响。方法选取2022年10月—2023年10月新疆维吾尔自治区人民医院泌尿中心诊治ED患者89例作为研究对象,随机数字表法分为单药组44例和联合组45例。单药组给予西地那非口服治疗,联合组在单药组基础上给予还原型谷胱甘肽片口服治疗,2组患者均连续治疗1个月。观察2组患者治疗前、治疗结束时及治疗后1个月的血管内皮功能(NO、ET、VEGF、ES)、炎性因子(hs-CRP、IL-6、IL-8、IL-10)、勃起功能(IIEF-5、QEQ、EHS、PSV)变化,比较2组临床疗效、不良事件发生率。结果治疗结束后1个月,联合组临床治疗总有效率为91.11%,高于单药组的75.00%(χ^(2)/P=4.121/0.042);治疗结束时及治疗结束后1个月,联合组患者血清NO、VEGF水平显著高于单药组,ET水平显著低于单药组(治疗结束时:t/P=5.323/<0.001,3.808/<0.001,3.683/<0.001;治疗结束后1个月:t/P=2.615/0.011,3.197/0.002,3.089/0.003);血清hs-CRP、IL-6水平显著低于单药组(治疗结束时:t/P=8.323/<0.001,2.364/0.020;治疗结束后1个月:t/P=6.787/<0.001,2.662/0.009);IIEF-5、QEQ、EHS及PSV均显著高于治疗前,其中EHS及PSV也显著高于单药组(治疗结束时:t/P=6.410/<0.001,4.066/<0.001;治疗结束后1个月:t/P=8.928/<0.001,4.532/<0.001);2组患者不良事件发生率比较差异无统计学意义(P>0.05)。结论谷胱甘肽联合西地那非能够有效改善ED患者血管内皮功能及勃起功能,同时显著降低患者炎性因子水平,且对于ED患者具有显著临床疗效及安全性。

以卡托普利为模板银纳米团簇的制备及生物硫醇的检测研究

生物硫醇在生物体内的代谢过程中起着重要作用,如体内半胱氨酸(Cys)和谷胱甘肽(GSH)含量异常会引发一系列疾病。为了建立一种快速、定量检测生物硫醇的荧光分析方法,本文以卡托普利(Captopril,Capt)为模板,采用湿化学还原法成功合成了发红色荧光的银纳米团簇(Capt-Ag NCs),对Capt-Ag NCs的形貌、结构、元素组成和光学性能等进行了表征。基于生物硫醇小分子对Capt-Ag NCs的荧光猝灭作用建立了定量检测Cys和GSH的新方法,在最佳实验条件下,Cys和GSH的线性范围分别为1~100μmol/L和2~180μmol/L,检出限分别为0.054μmol/L和0.23μmol/L。该方法可用于人血浆中Cys的测定,回收率为95.3%~103.3%。实验结果表明,CaptAg NCs作为一种新型信号关闭荧光探针,可用于Cys和GSH等生物硫醇的检测。

硫氢化钠增加高糖高脂条件下小鼠心房肌细胞系HL-1谷胱甘肽合成

目的研究硫氢化钠(NaHS)是否通过调节谷胱甘肽(GSH)的合成,降低活性氧自由基(ROS)产生,改善小鼠2型糖尿病心肌病(DCM)。方法将小鼠心房肌细胞系HL-1分为对照组、高葡萄糖(HG:40 mmol/L)和棕榈酸(Pal:500μmol/L)处理组;以及硫氢化钠(NaHS,100μmol/L)、DL-炔丙基甘氨酸[PPG,胱硫醚γ裂解酶(CSE)抑制剂,1 mmol/L]和N-乙酰-L-半胱氨酸(NAC,ROS抑制剂,5 mmol/L)处理72 h组。Western blot检测CSE和谷胱甘肽合成酶(GSS)的表达;二氢乙锭(DHE)和二氯氟甲烷(DCFH)检测ROS含量;免疫共沉淀检测核因子红细胞系2相关因子2(Nrf2)泛素化水平及Nrf2与肌肉特异性环指蛋白1(Murf1)的相互作用。结果与对照组比高糖高脂处理HL-1细胞后CSE、溶质载体家族7成员11(SLC7A11)、谷氨酸半胱氨酸连接酶催化亚基C(GCLC)、谷氨酸半胱氨酸连接酶修饰亚基M(GCLM)、谷胱甘肽合成酶(GSS)的表达水平下降,而NaHS能恢复其表达。高糖高脂组ROS含量高于NaHS组。与NaSH组比高糖高脂条件下Murf1与Nrf2的相互作用增加,Nrf2泛素化水平明显增加。结论硫氢化钠减轻Nrf2的泛素化,增加GSH合成关键酶的表达。