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周氏啮小蜂CcGSTS1基因的克隆·蛋白表达纯化及酶学特征分析
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作者 覃东玉 任睿 +2 位作者 孙美娣 李敏 潘丽娜 《安徽农业科学》 CAS 2024年第2期93-96,109,共5页
鉴定一个编码周氏啮小蜂谷胱甘肽S-转移酶序列的全长基因CcGSTS1,系统发育分析发现,该基因与丽蝇蛹集金小蜂NvGSTS6基因具有高同源性。体外表达、纯化的重组CcGSTS1可催化CDNB与GSH结合,最适反应pH为7.0。该试验为进一步研究周氏啮小蜂C... 鉴定一个编码周氏啮小蜂谷胱甘肽S-转移酶序列的全长基因CcGSTS1,系统发育分析发现,该基因与丽蝇蛹集金小蜂NvGSTS6基因具有高同源性。体外表达、纯化的重组CcGSTS1可催化CDNB与GSH结合,最适反应pH为7.0。该试验为进一步研究周氏啮小蜂CcGSTS1基因功能奠定基础。 展开更多
关键词 周氏啮小蜂 谷胱甘肽s-转移酶 系统发育分析 酶活
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GSTP1通过调控STAT3信号通路促进乳腺癌细胞增殖与阿霉素耐药
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作者 戴素华 嵇晓艳 戴夕超 《临床肿瘤学杂志》 CAS 2024年第2期114-119,共6页
目的 探讨谷胱甘肽S-转移酶P1(GSTP1)对人乳腺癌MCF-7细胞增殖与阿霉素耐药性的影响及其作用机制。方法 Western blotting检测野生型乳腺癌细胞MCF-7和阿霉素耐药乳腺癌细胞MCF-7/ADR中的GSTP1表达量,通过在MCF-7细胞中转染Flag-GSTP1... 目的 探讨谷胱甘肽S-转移酶P1(GSTP1)对人乳腺癌MCF-7细胞增殖与阿霉素耐药性的影响及其作用机制。方法 Western blotting检测野生型乳腺癌细胞MCF-7和阿霉素耐药乳腺癌细胞MCF-7/ADR中的GSTP1表达量,通过在MCF-7细胞中转染Flag-GSTP1质粒过表达GSTP1,在MCF-7/ADR细胞中转染GSTP1敲低慢病毒(shGSTP1)干扰GSTP1表达;平板克隆形成实验、CCK-8法和流式细胞术检测转染后乳腺癌细胞的增殖能力、阿霉素耐药性及凋亡水平的变化。Western blotting检测GSTP1表达水平的变化对STAT3通路激活的影响。结果 GSTP1在MCF-7细胞中表达量极低,且显著低于MCF-7/ADR细胞系。GSTP1过表达的MCF-7/ADR细胞克隆形成数量显著多于野生型MCF-7细胞(P<0.05)。过表达GSTP1显著提升了MCF-7细胞的增殖能力,而在MCF-7/ADR细胞系干扰GSTP1表达水平后显著抑制了细胞增殖能力(P<0.05)。CCK-8结果显示,在0.1、1、10、50μmol/ml不同浓度的阿霉素处理下,GSTP1表达水平与MCF-7细胞对阿霉素的耐药性呈正相关(P<0.05)。流式细胞术检测结果显示,GSTP1过表达组(Flag-GSTP1)和对照组(Flag)的凋亡率分别为(11.41±1.16)%和(21.1±1.72)%,GSTP1过表达显著抑制了阿霉素诱导的细胞凋亡(P<0.05)。Western blotting检测结果显示,GSTP1的过表达激活了STAT3信号通路,同时在MCF-7/ADR细胞系中抑制STAT3显著降低了GSTP1的表达水平(P<0.05)。结论 GSTP1通过上调STAT3表达调节乳腺癌细胞MCF-7的增殖能力和对阿霉素的耐药性。 展开更多
关键词 乳腺癌 谷胱甘肽s转移酶P1 增殖 阿霉素耐药
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山新杨谷胱甘肽S-转移酶编码基因PdbGSTU功能分析
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作者 黄颖 王晓东 遇文婧 《北京林业大学学报》 CAS CSCD 北大核心 2024年第8期68-78,共11页
【目的】本研究旨在通过分析山新杨谷胱甘肽S-转移酶编码基因PdbGSTU的抗病功能,为林木抗性育种提供基因资源与抗性种质。【方法】克隆PdbGSTU基因序列并对其进行生物信息学分析,利用荧光定量PCR技术分析该基因的组织特异性表达及植物... 【目的】本研究旨在通过分析山新杨谷胱甘肽S-转移酶编码基因PdbGSTU的抗病功能,为林木抗性育种提供基因资源与抗性种质。【方法】克隆PdbGSTU基因序列并对其进行生物信息学分析,利用荧光定量PCR技术分析该基因的组织特异性表达及植物激素诱导下的表达模式。利用转基因技术获得山新杨的过/抑制表达PdbGSTU基因植株,通过观察比较接种细链格孢菌后各植株叶片的表型和病斑面积,验证该基因的抗病功能;同时测定接种病原菌前后,野生型和转基因山新杨植株内过氧化氢含量和抗氧化相关酶活性。【结果】(1)山新杨PdbGSTU基因开放阅读框全长753 bp,编码氨基酸250个,对应的蛋白质相对分子质量为29.01 kDa,为稳定的酸性亲水蛋白,定位于细胞质中;系统进化分析显示,PdbGSTU蛋白与银中杨的蛋白KAJ6918316亲缘关系最近;启动子序列分析显示,PdbGSTU基因启动子序列含多种响应植物激素和逆境胁迫的顺式作用元件。(2)RT-qPCR结果显示,PdbGSTU基因在山新杨顶芽表达量最高,在其根部表达量最低,且该基因受茉莉酸甲酯、水杨酸和1-氨基环丙基-1-羧酸3种植物激素诱导,均上调表达。(3)接种细链格孢菌后,野生型和抑制表达PdbGSTU基因植株的叶片上,病斑面积分别为6.42和16.46 mm2,而过表达PdbGSTU基因的植株叶片上,少部分接种点出现明显病斑,其余接种部分仅出现褪色。【结论】PdbGSTU正向参与山新杨对细链格孢菌侵染的抵御过程,可通过清除活性氧提高杨树对病原菌的抗性。 展开更多
关键词 基因表达 山新杨 谷胱甘肽s-转移酶 细链格孢菌 抗病功能
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重金属汞胁迫下菲律宾蛤仔GPx和GST基因表达分析
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作者 郑志龙 闫路路 +1 位作者 闫喜武 秦艳杰 《安徽农业科学》 CAS 2024年第10期113-119,共7页
为研究重金属汞胁迫下菲律宾蛤仔谷胱甘肽过氧化物酶(glutathione peroxidase,GPx)和谷胱甘肽-S-转移酶(glutathione-S-transferase,GST)基因的表达情况,用汞对指示生物菲律宾蛤仔(Ruditapes philippinarum)进行单一慢性暴露试验,分别在... 为研究重金属汞胁迫下菲律宾蛤仔谷胱甘肽过氧化物酶(glutathione peroxidase,GPx)和谷胱甘肽-S-转移酶(glutathione-S-transferase,GST)基因的表达情况,用汞对指示生物菲律宾蛤仔(Ruditapes philippinarum)进行单一慢性暴露试验,分别在0、12、24 h以及2、3、4、5、6、7和8 d时检测菲律宾蛤仔内脏团和鳃中GPx和GST基因的表达情况。结果表明,菲律宾蛤仔内脏团中GPx和GST基因的表达量都呈现波动变化趋势,分别在24和12 h时表达量最高(P<0.05),6 d时GPx基因表达量最低(P>0.05),3 d时GST基因表达量最低(P<0.05);GPx基因在鳃中的表达量在8 d时最高(P<0.05);鳃中GST基因的表达量在5 d时最高(P<0.05)。以上结果表明汞暴露在短期内能够诱导GPx和GST基因进行不同程度的表达,但其随着时间的延长呈明显的抑制作用。该研究结果为揭示重金属汞对菲律宾蛤仔的毒性机理提供了理论依据。 展开更多
关键词 重金属汞 菲律宾蛤仔 谷胱甘肽过氧化物酶 谷胱甘肽-s-转移酶
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山新杨谷胱甘肽S-转移酶PdbGST基因的克隆与胁迫表达分析
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作者 平晓帆 遇文婧 黄颖 《林业科技》 2024年第3期33-40,共8页
在植物中,谷胱甘肽S-转移酶是一类多功能酶,能够在其响应非生物胁迫中发挥作用。为研究该基因在杨树中的功能,本研究克隆了山新杨谷胱甘肽S-转移酶基因Pdb GST及其启动子序列,对其进行生物信息学分析和表达特征分析。结果显示,Pdb GST... 在植物中,谷胱甘肽S-转移酶是一类多功能酶,能够在其响应非生物胁迫中发挥作用。为研究该基因在杨树中的功能,本研究克隆了山新杨谷胱甘肽S-转移酶基因Pdb GST及其启动子序列,对其进行生物信息学分析和表达特征分析。结果显示,Pdb GST基因的开放阅读框(ORF)全长666 bp,编码221个氨基酸,其形成的蛋白质相对分子质量为25.57 k Da,为稳定亲水性酸性蛋白,定位于细胞质中。系统进化分析表明,山新杨Pdb GST蛋白与山新杨中的XP_034926648蛋白的同源性最高。对克隆获取的2 000 bp启动子序列进行分析,结构显示该启动子序列中,含有多种响应非生物胁迫的顺式作用元件。荧光定量PCR分析显示Pdb GST基因在山新杨根部表达量最高,其次在山新杨叶部也有较高表达量,相反在山新杨顶芽中表达量最低。此外,对Pdb GST基因响应非生物胁迫分析显示,山新杨根部Pdb GST基因受诱导后即出现持续且大量的表达。 展开更多
关键词 山新杨 谷胱甘肽s-转移酶 非生物胁迫 表达模式
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Glutathione S-transferase(GST) gene expression profiles in two marine bivalves exposed to BDE-47 and their potential molecular mechanisms 被引量:2
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作者 李斐 吴惠丰 +2 位作者 王清 李雪花 赵建民 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2015年第3期705-713,共9页
Glutathione S-transferases(GSTs) are phase II enzymes that facilitate the detoxification of xenobiotics and play important roles in antioxidant defense. We investigated the expression patterns of seven Venerupis phili... Glutathione S-transferases(GSTs) are phase II enzymes that facilitate the detoxification of xenobiotics and play important roles in antioxidant defense. We investigated the expression patterns of seven Venerupis philippinarum GSTs( Vp GSTs) and four Mytilus galloprovincialis GSTs( Mg GSTs) following exposure to BDE-47. Differential expressions of the seven V p GSTs and four Mg GSTs transcripts were observed,with differences between the hepatopancreas and gills. Among these GSTs,the sigma classes( Vp GSTS1,Vp GSTS2,Vp GSTS3,Mg GST1,and Mg GST3) were highly expressed in response to BDE-47 exposure,demonstrating their potential as molecular biomarkers for environmental biomonitoring studies. We obtained the three-dimensional crystal structures of V p GSTs and Mg GSTs by homologous modeling. A model to elucidate the binding interactions between the ligands and receptors was defined by molecular docking. Hydrophobic and π were the most often observed interactions between BDE-47 and the GSTs. 展开更多
关键词 谷胱甘肽s-转移酶 gsts 基因表达谱 分子机制 海洋贝类 环境生物监测 三维晶体结构 分子标志物
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Glutathione-S-transferase (GSTM1,GSTT1) and the risk of gastrointestinal cancer in a Korean population
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作者 Jin-Mei Piao Min-Ho Shin +8 位作者 Sun-Seog Kweon Hee Nam Kim Jin-Su Choi Woo-Kyun Bae Hyun-Jeong Shim Hyeong-Rok Kim Young-Kyu Park Yoo-Duk Choi Soo-Hyun Kim 《World Journal of Gastroenterology》 SCIE CAS CSCD 2009年第45期5716-5721,共6页
AIM: To evaluate the association of glutathione S-transferase mu (GSTM1) and glutathione S-transferase theta (GSTT1) null genotypes with the risk of gastric cancer (GC) and colorectal cancer (CRC) in a South Korean po... AIM: To evaluate the association of glutathione S-transferase mu (GSTM1) and glutathione S-transferase theta (GSTT1) null genotypes with the risk of gastric cancer (GC) and colorectal cancer (CRC) in a South Korean population.METHODS: We conducted a population-based, largescale case-control study including 2213 GCs, 1829 CRCs, and 1699 controls. Null and non-null genotypesof GSTM1 and GSTT1 were determined using realtime PCR.RESULTS: The null genotypes of GSTM1 and GSTT1 were not signifi cantly associated with elevated risk of gastric (OR = 1.070, 95% CI = 0.935-1.224; OR = 1.101, 95% CI = 0.963-1.259, respectively) or colorec-tal cancer (OR = 1.065, 95% CI = 0.923-1.228; OR = 1.041, 95% CI = 0.903-1.200, respectively). The frequency of the combined null GST genotype was not different between the two cancer groups and controls. Moreover, smoking, drinking, and age did not modify the association between these genotypes and the risk of gastric or colorectal cancer.CONCLUSION: GSTM1 and GSTT1 null genotypes were not associated with increased risk of GC or CRC in Koreans. 展开更多
关键词 gstM1基因 谷胱甘肽s 胃肠道肿瘤 韩国人 转移酶 风险 人口 聚合酶链反应
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Modification of the Genetic Polymorphism of Glutathione-S-Transferase (GSTM1 and GSTT1) in Motorcycle Drivers Exposed to BTEX in Cotonou
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作者 Cachon Fresnel Boris Sagbo H. Firmin +4 位作者 Nanoukon Chimène Djedatin Gustave Atindehou Ménonvè Senou Maximin Ayi-Fanou Lucie 《Journal of Biosciences and Medicines》 CAS 2022年第11期104-112,共9页
The glutathione-S-transferase genes mainly the GSTM1 and GSTT1 alleles are responsible for the synthesis of detoxication enzymes that can remove toxic substances. The objective of this study is to seek changes in the ... The glutathione-S-transferase genes mainly the GSTM1 and GSTT1 alleles are responsible for the synthesis of detoxication enzymes that can remove toxic substances. The objective of this study is to seek changes in the genetic polymorphism of glutathione-S-transferase GSTM1 and GSTT1 in motorcycle drivers exposed to BTEX. Our study group consists of 60 motorcycle drivers including 30 professional and 30 non-professional. Blood samples were preleveled from the study population in the EDTA tubes and DNA was extracted by the phenol/chloroform method. The PCR technique was used to determine the presence or absence of genes. Our results showed that the percentage of GSTM1 null genotype has a statistically significant difference (P = 0.02), while the percentage of GSTT1 null genotype was non-significant (P = 0.76) between the two groups. The percentage of deletion of both genes is higher in professional than non-professional motorcycle drivers. Air pollution in Cotonou by BTEX seems to influence the deletion of the GSTM1 and GSTT1 genes at a higher percentage among professional than non-professional motorcycle drivers. 展开更多
关键词 BTEX glutathione-s-transferase Deletion PCR Motorcycle Drivers
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Carboxylesterase and Glutathione-S-Transferase (GST's) Induced Resistance to Bacillus thuringiensis Toxin CrylAb in Rice Leaf Folder, Cnaphalocrocis medinalis (Guenee) Populations
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作者 Veegala Ramesh Babu Vemuri Shashi Bhushan +3 位作者 Chintalapati Padmavathy: Muthugonder Mohan SenaMahendran. Balachandran Bellamkonda Ramesh 《Journal of Agricultural Science and Technology(A)》 2013年第1期53-59,共7页
关键词 谷胱甘肽-s-转移酶 CRY1AB杀虫蛋白 稻纵卷叶螟 羧酸酯酶 杀虫毒素 苏云金芽孢杆菌 gst 诱导抗性
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EphA2蛋白与GST-pi蛋白在结直肠癌组织中表达及其预后意义
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作者 陆军 张艳皊 +1 位作者 陈洁 张岩 《国际检验医学杂志》 CAS 2023年第5期541-547,共7页
目的探讨促红细胞生成素肝细胞A2(EphA2)蛋白与谷胱苷肽S-转移酶Pi(GST-pi)蛋白在结直肠癌标本中的表达及其与临床病理特征、预后的关系。方法收集2014年6月至2020年2月于该院接受手术治疗的114例结直肠癌患者,应用免疫组化染色技术检测... 目的探讨促红细胞生成素肝细胞A2(EphA2)蛋白与谷胱苷肽S-转移酶Pi(GST-pi)蛋白在结直肠癌标本中的表达及其与临床病理特征、预后的关系。方法收集2014年6月至2020年2月于该院接受手术治疗的114例结直肠癌患者,应用免疫组化染色技术检测EphA2蛋白与GST-pi蛋白在癌组织及其对应癌旁组织中的表达,分析二者与结直肠癌临床病理特征及总生存率的关系。结果EphA2蛋白与GST-pi蛋白的阳性表达率分别为54.4%和62.3%,显著高于二者在癌旁组织中的表达,差异有统计学意义(P<0.05)。与EphA2蛋白阴性表达患者相比,EphA2蛋白阳性表达患者的肿瘤最大径>5 cm占比、有淋巴结转移占比、有淋巴血管侵犯占比更高;GST-pi蛋白表达与结直肠癌患者的肿瘤最大径及T分期有关(P<0.05)。Kaplan-Meier生存分析表明EphA2蛋白阳性表达与阴性表达患者5年总生存率分别为20.1%和68.8%,差异有统计学意义(χ^(2)=16.973,P<0.001);GST-pi蛋白阳性表达与阴性表达者的5年总生存率分别为28.2%和61.8%,差异有统计学意义(χ^(2)=3.974,P=0.046)。单、多变量Cox回归分析表明,EphA2蛋白阳性表达(风险比=3.442,95%CI:1.722~6.881,P<0.001)、GST-pi蛋白阳性表达(风险比=2.203,95%CI:1.088~4.460,P=0.028)、淋巴结转移(风险比=3.627,95%CI:1.699~7.742,P=0.001)及淋巴血管侵犯(风险比=2.113,95%CI:1.033~4.322,P=0.040)是结直肠癌患者预后不佳的独立预测因素。结论EphA2蛋白与GST-pi蛋白或许可作为结直肠癌患者预后评估的重要参考指标。 展开更多
关键词 结直肠癌 促红细胞生成素肝细胞A2 谷胱苷肽s-转移酶Pi 临床病理特征 预后
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甘蔗谷胱甘肽硫转移酶ScGSTF1与P3N-PIPO互作应答甘蔗花叶病毒侵染的研究
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作者 杨宗桃 焦文迪 +6 位作者 张海 张克闽 程光远 罗廷绪 曾康 周营栓 徐景升 《作物学报》 CAS CSCD 北大核心 2023年第10期2665-2676,共12页
谷胱甘肽S-转移酶(glutathione S-transferase, GST, EC2.5.1.18)广泛分布于具有细胞结构的生物中。在植物中,GST参与调控生长发育、异生物质解毒及逆境应答。本课题组以甘蔗花叶病毒(Sugarcanemosaicvirus,SCMV)编码蛋白P3N-PIPO为诱饵... 谷胱甘肽S-转移酶(glutathione S-transferase, GST, EC2.5.1.18)广泛分布于具有细胞结构的生物中。在植物中,GST参与调控生长发育、异生物质解毒及逆境应答。本课题组以甘蔗花叶病毒(Sugarcanemosaicvirus,SCMV)编码蛋白P3N-PIPO为诱饵,从甘蔗(Saccharumspp.hybrid)叶片cDNA酵母文库中筛选到1个GST基因,并从甘蔗原始栽培种Badila中克隆了该基因,其开放读码框(open reading frame, ORF)长度为645 bp,编码214个氨基酸。序列比对及进化树分析表明,该基因编码蛋白属于GST家族的Phi (F)类型(GSTF),因此命名为ScGSTF1。进一步的生物信息学分析表明,ScGSTF1不存在跨膜区域,为稳定的亲水性脂溶蛋白;GST蛋白在单子叶和双子叶植物中及C3和C4植物中存在明显的分化。酵母双杂交(yeast two-hybrid, Y2H)和双分子荧光互补(bimolecular fluorescence complementation, BiFC)试验表明, ScGSTF1与SCMV-P3N-PIPO蛋白互作。亚细胞定位试验表明ScGSTF1定位于内质网。实时荧光定量PCR分析表明, ScGSTF1在甘蔗各个组织中显著差异表达,在第3节间中的表达量最高,在心叶及根中的表达量最低;SCMV侵染显著性影响ScGSTF1基因的表达水平,侵染早期上调表达,随后下调,但仍保持显著高于对照的水平。 展开更多
关键词 甘蔗 甘蔗花叶病毒 谷胱甘肽硫转移酶 P3N-PIPO 蛋白互作
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割手密GST基因家族的分析及其对阿特拉津的响应
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作者 林焕泰 薛耀威 +1 位作者 高三基 王锦达 《福建农林大学学报(自然科学版)》 CSCD 北大核心 2023年第5期582-590,共9页
从全基因组的角度分析了割手密谷胱苷肽⁃S⁃转移酶(GST)基因家族成员的特征,共鉴定出129个GST。通过对割手密GST蛋白的系统发育分析,发现这些GST可以划分为4个亚家族,且每个亚家族内的蛋白排列紧凑,说明同亚族内部基因进化关系均比较亲... 从全基因组的角度分析了割手密谷胱苷肽⁃S⁃转移酶(GST)基因家族成员的特征,共鉴定出129个GST。通过对割手密GST蛋白的系统发育分析,发现这些GST可以划分为4个亚家族,且每个亚家族内的蛋白排列紧凑,说明同亚族内部基因进化关系均比较亲近。此外,对在阿特拉津处理下GST基因家族中10个F亚族基因的表达水平进行检测,结果表明,根中的SpGSTF1、SpGSTF9、SpGSTF14、SpGSTF16、SpGSTF18、SpGSTF31在阿特拉津胁迫下表达显著上调,推测这6个基因可能在响应阿特拉津胁迫的过程中起重要作用。研究结果有助于揭示割手密GST家族的进化和功能演替,并对进一步了解割手密GST对阿特拉津的解毒响应机制提供理论参考,同时也为培养抗除草剂甘蔗品种提供基因资源。 展开更多
关键词 割手密 谷胱甘肽⁃s⁃转移酶 基因家族分析 阿特拉津
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GSTO1抑制TGFβ诱导的大鼠心脏成纤维细胞增殖和活化
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作者 张彤 谢赛阳 +1 位作者 邓伟 唐其柱 《中国病理生理杂志》 CAS CSCD 北大核心 2023年第4期656-662,共7页
目的:研究谷胱甘肽S-转移酶ω1(GSTO1)在转化生长因子β(TGFβ)诱导心脏成纤维细胞增殖与活化中的作用。方法:分离乳大鼠心脏成纤维细胞并进行体外培养。采用含绿色荧光蛋白(GFP)的重组腺病毒质粒构建对照(Ad-GFP)或GSTO1过表达(Ad-GST... 目的:研究谷胱甘肽S-转移酶ω1(GSTO1)在转化生长因子β(TGFβ)诱导心脏成纤维细胞增殖与活化中的作用。方法:分离乳大鼠心脏成纤维细胞并进行体外培养。采用含绿色荧光蛋白(GFP)的重组腺病毒质粒构建对照(Ad-GFP)或GSTO1过表达(Ad-GSTO1)质粒并转染心脏成纤维细胞,再用大鼠TGFβ(rTGFβ)刺激细胞24 h,实验分为4组:Ad-GFP+PBS、Ad-GFP+rTGFβ、Ad-GSTO1+PBS和Ad-GSTO1+rTGFβ。运用qPCR和免疫荧光染色确定转染的有效性;Western blot检测GSTO1蛋白表达;划痕实验和EdU掺入实验评估细胞迁移和增殖;CCK-8实验评估细胞活力;Western blot和细胞免疫荧光染色检测心脏成纤维细胞活化标志物α-平滑肌肌动蛋白(α-SMA)、I型胶原(Col I)和纤连蛋白(FN)的表达水平,并检测心脏成纤维细胞内P38 MAPK和Smad3信号通路相关蛋白水平。结果:与Ad-GFP+rTGFβ组相比,GSTO1过表达抑制TGFβ诱导的心脏成纤维细胞增殖(P<0.05),降低TGFβ刺激后α-SMA、Col I和FN的表达水平(P<0.05),抑制P38 MAPK/Smad3信号通路激活(P<0.05)。结论:GSTO1过表达通过抑制P38 MAPK/Smad3信号通路而阻遏TGFβ诱导的大鼠心脏成纤维细胞增殖和活化。 展开更多
关键词 心肌纤维化 谷胱甘肽s-转移酶ω1 心脏成纤维细胞 转化生长因子β 细胞增殖
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星豹蛛两个GST基因克隆、鉴定及其对溴氰菊酯胁迫的响应表达
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作者 焦丽亚琳 赵萌萌 +3 位作者 王美 牛越 王西 李锐 《昆虫学报》 CAS CSCD 北大核心 2023年第9期1161-1170,共10页
【目的】为明确星豹蛛Pardosa astrigera体内2个谷胱甘肽S-转移酶(glutathione S-transferases,GSTs)基因的时空表达模式,并探究其是否能对溴氰菊酯的胁迫做出响应。【方法】基于星豹蛛转录组数据库,PCR克隆星豹蛛GST基因PaGSTd1和PaGS... 【目的】为明确星豹蛛Pardosa astrigera体内2个谷胱甘肽S-转移酶(glutathione S-transferases,GSTs)基因的时空表达模式,并探究其是否能对溴氰菊酯的胁迫做出响应。【方法】基于星豹蛛转录组数据库,PCR克隆星豹蛛GST基因PaGSTd1和PaGSTd2全长cDNA序列并进行生物信息学分析;利用RT-qPCR检测PaGSTd1和PaGSTd2在星豹蛛不同发育阶段(2-6龄若蛛和成蛛)、雌雄成蛛不同组织(头胸部、腹和足)以及通过药膜法利用不同浓度[LC 10(5.151 mg/L),LC 30(8.619 mg/L)和LC 50(12.311 mg/L)]溴氰菊酯胁迫不同时间(6,12,18,24和48 h)雄成蛛中的表达量。【结果】星豹蛛PaGSTd1(GenBank登录号:OR096398)全长cDNA序列长708 bp,开放阅读框长645 bp,编码214个氨基酸;星豹蛛PaGSTd2(GenBank登录号:OR096399)全长cDNA序列长793 bp,开放阅读框长645 bp,编码214个氨基酸。RT-qPCR检测结果表明,PaGSTd1和PaGSTd2在星豹蛛不同发育阶段和成蛛不同组织都有表达,均在5龄若蛛中的表达量最低;PaGSTd1在4龄若蛛中的表达量最高,PaGSTd2在成蛛中的表达量最高;PaGSTd1和PaGSTd2在足中的表达量最高,在除腹部外的其他组织中表达量均为雄成蛛的显著高于雌成蛛的。LC 10溴氰菊酯胁迫24 h时星豹蛛雄成蛛中PaGSTd1被诱导表达,6和18 h时PaGSTd2被诱导表达;LC 30溴氰菊酯胁迫18,24和48 h时雄成蛛中PaGSTd1被诱导表达,18 h时PaGSTd2被诱导表达;LC 50溴氰菊酯胁迫24 h时雄成蛛中PaGSTd1和PaGSTd2被诱导表达。【结论】本研究克隆得到星豹蛛两个GST基因PaGSTd1和PaGSTd2,均在星豹蛛足中表达量最高,说明足中这2个GST基因在对外源物质的解毒起到重要作用;这两个GST基因可以被溴氰菊酯诱导表达,说明可能参与星豹蛛对溴氰菊酯的胁迫响应,为后续GSTs的功能研究提供线索。 展开更多
关键词 星豹蛛 谷胱甘肽s-转移酶(gst) 溴氰菊酯 基因表达
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GSTM5在骨髓增生异常综合征细胞中的表达及意义 被引量:1
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作者 常青 董腾腾 +3 位作者 虞亚菲 张兆怡 王驰 李绵洋 《国际检验医学杂志》 CAS 2023年第9期1035-1038,共4页
目的探讨谷胱甘肽S-转移酶Mu 5(GSTM5)基因在骨髓增生异常综合征(MDS)细胞SKM-1中的表达及意义。方法采用去甲基化药物地西他滨(DAC)、GSTM5过表达质粒(SKM-1ovGSTM5)和谷胱甘肽抑制剂丁硫氨酸亚砜亚胺(BSO)分别处理SKM-1细胞。采用实... 目的探讨谷胱甘肽S-转移酶Mu 5(GSTM5)基因在骨髓增生异常综合征(MDS)细胞SKM-1中的表达及意义。方法采用去甲基化药物地西他滨(DAC)、GSTM5过表达质粒(SKM-1ovGSTM5)和谷胱甘肽抑制剂丁硫氨酸亚砜亚胺(BSO)分别处理SKM-1细胞。采用实时荧光定量PCR(RT-qPCR)检测GSTM5 mRNA表达水平,Western blotting检测GSTM5蛋白表达水平,CCK-8试验检测SKM-1细胞的增殖能力,流式细胞术检测SKM-1细胞凋亡率和细胞周期分布,另采用谷胱甘肽(GSH)检测试剂盒测定GSH水平。结果与对照相比,过表达GSTM5质粒后SKM-1细胞增殖能力降低(P<0.05),而细胞凋亡率和细胞周期分布情况差异无统计学意义(P>0.05)。DAC处理及过表达GSTM5的SKM-1细胞GSH水平均显著降低(P<0.05)。与对照相比,BSO处理SKM-1细胞后,细胞增殖能力显著降低(P<0.05)。结论GSTM5可通过调控GSH水平抑制SKM-1细胞的增殖,并具有作为MDS诊断标志物的潜力。 展开更多
关键词 骨髓增生异常综合征 谷胱甘肽s-转移酶Mu 5 甲基化 谷胱甘肽
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Polymorphism of glutathione S-transferase mu 1 and theta 1 genes and hepatocellular carcinoma in southern Guangxi, China 被引量:8
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作者 Zhuo-LinDeng Yi-PingWei YunMa 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第2期272-274,共3页
AIM: Glutathione S-transferase mu 1 (GSTM1) and theta 1(GSTT1) genes are involved in the metabolism of a wide range of carcinogens, but deletions of the genes are commonly found in the population. The present study wa... AIM: Glutathione S-transferase mu 1 (GSTM1) and theta 1(GSTT1) genes are involved in the metabolism of a wide range of carcinogens, but deletions of the genes are commonly found in the population. The present study was undertaken to evaluate the association between GSTM1 and GSTT1 gene polymorphisms and hepatocellular carcinoma (HCC) risk.METHODS: The genetic polymorphisms were studied at an aflatoxin highly contaminated region in Guangxi, China.Polymerase chain reaction (PCR) technique was used to detect the presence or absence of the GSTM1 and GSTT1 genes in blood samples. The case group was composed of 181 patients of HCC identified by the pathologists and the control group was composed of 360 adults without any tumor.RFSULTS: The frequencies of GSTM1 and GSTT1 null genotypes in the control were 47.8% and 42.7%, while those in the HCC group were 64.6% and 59.7%, respectively. The differences between HCC group and control group were very significant (P<0.01). GSTM1 and GSTT1 combined null genotypes in HCC group and control group were 38.2% and 18.5%respectively, and the difference was significant (P<0.05).CONCLUSION: The GSTM1 and GSTT1 null genotypes are associated with an increased risk of HCC in a special geographic environment. Combination of the two null genotypes in an individual is substantially increased twice the risk of HCC. 展开更多
关键词 多态现象 谷胱甘肽 s-转移酶 μ1基因 θ1基因 肝细胞癌 肿瘤 消化系统 中国 广西
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Alterations of glutathione S-transferase and matrix metalloproteinase-9 expressions are early events in esophageal carcinogenesis 被引量:37
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作者 Laszlo Herszenyi Istvan Hritz +4 位作者 Istvan Pregun Ferenc Sipos Mark Juhasz Bela Molnar Zsolt Tulassay 《World Journal of Gastroenterology》 SCIE CAS CSCD 2007年第5期676-682,共7页
AIM: To investigate the role of glutathione S-transferase (GST) and matrix metalloproteinase-9 (MMP-9) expres-sions in the development and progression of reflux es-ophagitis-Barrett’s metaplasia-dysplasia-adenocarcin... AIM: To investigate the role of glutathione S-transferase (GST) and matrix metalloproteinase-9 (MMP-9) expres-sions in the development and progression of reflux es-ophagitis-Barrett’s metaplasia-dysplasia-adenocarcinoma sequence in the esophagus.METHODS: GST and MMP-9 expressions were analyzed in 51 paraffin-embedded tissue samples by immunohisto-chemistry including patients with reflux esophagitis (n = 7), Barrett’s metaplasia (n = 14), Barrett and esophagi-tis (n = 8), Barrett and dysplasia (n = 7), esophageal adenocarcinoma (n = 8) and a control group without any histological changes (n = 7). Immunostaining was determined semiquantitatively. Statistical analysis with one-way ANOVA, LSD test and correlation analysis were performed. P value of < 0.05 was considered significant.RESULTS: GST expression was significantly higher while MMP-9 expression was significantly lower in control group compared to Barrett’s metaplasia and the other groups. No major changes were observed between Bar-rett, esophagitis, and Barrett and concomitant esophagi-tis. Barrett and concomitant dysplasia, and adenocarci-noma revealed a significant lower expression of GST and higher levels of MMP-9 compared to all other groups. Adenocarcinoma showed almost no expression of GST and significantly higher levels of MMP-9 than Barrett and concomitant dysplasia. Alterations of GST and MMP-9 were inversely correlated (r = - 0.82).CONCLUSION: Decreased GST and increased ex-pression of MMP-9 in Barrett’s metaplasia-dysplasia-adenocarcinoma sequence as compared to normal tissue suggest their association with esophageal tumorigenesis. Loss of GST and gain of MMP-9 in Barrett with dyspla-sia compared to non-dysplastic metaplasia indicate that these alterations may be early events in carcinogenesis. Quantification of these parameters in Barrett’s esopha-gus might be useful to identify patients at higher risk for progression to cancer. 展开更多
关键词 谷胱甘肽s-转移酶 基质金属蛋白酶-9 表达变化 食管癌 早期 肿瘤发生
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Response of Glutathione and Glutathione S-transferase in Rice Seedlings Exposed to Cadmium Stress 被引量:13
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作者 ZHANG Chun-hua GE Ying 《Rice science》 SCIE 2008年第1期73-76,共4页
A hydroponic culture experiment was done to investigate the effect of Cd stress on glutathione content (GSH) and glutathione S-transferase (GST, EC 2.5.1,18) activity in rice seedlings. The rice growth was severel... A hydroponic culture experiment was done to investigate the effect of Cd stress on glutathione content (GSH) and glutathione S-transferase (GST, EC 2.5.1,18) activity in rice seedlings. The rice growth was severely inhibited when Cd level in the solution was higher than 10 mg/L. In rice shoots, GSH content and GST activity increased with the increasing Cd level, while in roots, GST was obviously inhibited by Cd treatments, Compared with shoots, the rice roots had higher GSH content and GST activity, indicating the ability of Cd detoxification was much higher in roots than in shoots. There was a significant correlation between Cd level and GSH content or GST activity, suggesting that both parameters may be used as biomarkers of Cd stress in rice. 展开更多
关键词 RICE cadmium stress glutathione glutathione s-transferase enzyme activity
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BaP通过AhR和Nrf2信号通路对HepG2细胞中GSTP1的影响 被引量:1
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作者 王蕾 张梦迪 +6 位作者 苏敏 王朝杰 胡玉霞 白图雅 吕晓丽 李君 常福厚 《内蒙古医科大学学报》 2023年第3期235-239,256,共6页
目的探究苯并芘(benzo[a]pyrene,BaP)通过芳香烃受体(aryl hydrocarbon receptor,AhR)和核因子E2相关因子(subcellular localization of nuclear factor E2-related factor 2,Nrf2)信号通路对HepG2细胞中谷胱甘肽-S-转移酶P1(glutathion... 目的探究苯并芘(benzo[a]pyrene,BaP)通过芳香烃受体(aryl hydrocarbon receptor,AhR)和核因子E2相关因子(subcellular localization of nuclear factor E2-related factor 2,Nrf2)信号通路对HepG2细胞中谷胱甘肽-S-转移酶P1(glutathione s-transferase P1,GSTP1)的影响。方法将体外培养HepG2细胞分为Control组、BaP(10μmol/L)组、BaP(10μmol/L)+AhR抑制剂(1μmol/L)组和BaP(10μmol/L)+Nrf2抑制剂(1μmol/L)组。BaP组给予10μmol/L BaP培养24 h,抑制剂组给予1μmol/L抑制剂0.5 h后,加入10μmol/L BaP培养24 h,采用Western Blot和qPCR实验方法检测各组AhR、Nrf2、细胞色素P450s(cytochrome P450s,CYPs)、血红素加氧酶-1(heme oxygenase-1,HO-1)及GSTP1基因和蛋白的表达。结果与Control组相比,BaP组中AhR、Nrf2、GSTP1、CYP1A1及HO-1的mRNA和蛋白表达均升高(P<0.05);与BaP组相比,BaP+AhR抑制剂组中AhR、GSTP1和CYP1A1的mRNA和蛋白表达均降低(P<0.05),BaP+Nrf2抑制剂组中Nrf2、GSTP1和HO-1的表达均降低(P<0.05);与BaP+AhR抑制剂组相比,BaP+Nrf2抑制剂组GSTP1 mRNA和蛋白的表达降低,差异有统计学意义(P<0.05)。结论BaP通过AhR和Nrf2两条信号通路增加GSTP1的表达,以Nrf2信号通路为主导影响GSTP1的表达。 展开更多
关键词 苯并芘 芳香烃受体 核因子E2相关因子2 谷胱甘肽-s-转移酶P1 药物代谢酶
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Polymorphisms of the oxidant enzymes glutathione S-transferase and glutathione reductase and their association with resistance of Plasmodium falciparum isolates to antimalarial drugs 被引量:3
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作者 Raewadee Wisedpanichkij Wanna Chaicharoenkul +2 位作者 Poonuch Mahamad Prapichaya Prompradit Kesara Na-Bangchang 《Asian Pacific Journal of Tropical Medicine》 SCIE CAS 2010年第9期673-677,共5页
Objective:To investigate the association between amplification of the two regulatory genes controlling glutathione(GSH) levels,glutathione reductase(PfGR) and glutathione S-transferase (PfGST) genes and sensitivity of... Objective:To investigate the association between amplification of the two regulatory genes controlling glutathione(GSH) levels,glutathione reductase(PfGR) and glutathione S-transferase (PfGST) genes and sensitivity of Plasmodium falciparum(P.falciparum) isolates collected from different malaria endemic areas of Thailand to standard antimalarial drugs.Methods:A total of 70 P.falciparum isolates were collected from endemic areas of multi-drug resistance (Tak,Chantaburi and Ranong Provinces) during the year 2008-2009.The in vitro assessment of antimalarial activity of P.falciparum clones(K1- and Dd2 chloroquine resistant and 3D7- chloroquine sensitive) and isolates to chloroquine,quinine,mefloquine and arteusnate was performed based on SYBR Green modified assay.Results:68(97.14%),11(15.71%) and 28(40%) isolates respectively were classified as chloroquine-,quinine- and mefloquine-resistant isolates. With this limited number of P.falciparum isolates included in the analysis,no significant association between amplification of PfGST gene and sensitivity of the parasite to chloroquine, quinine,mefloquine and quinine was found.Based on PCR analysis,Dd2,Kl and 3D7 clones all contained only one copy of the PfGST gene.All isolates(70) also carried only one copy number of PfGST gene.There appears to be an association between amplification of PfGR gene and chloroquine resistance.The 3D7 and Dd2 clones were found to carry only one PfGR gene copy, whereas the K1 clone carried two gene copies.Conclusions:Chloroquine resistance is likely to be a consequence of multi-factors and enzymes in the GSH system may be partly involved. Larger number of parasite isolates are required to increase power of the hypothesis testing in order to confirm the involvement of both genes as well as other genes implicated in glutathione metabolism in conferring chloroquine resistance. 展开更多
关键词 PLAsMODIUM FALCIPARUM Drug REsIsTANCE glutathione reductase(PfGR) glutathione s-transferase(P/gst)
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