芥菜BjGSTF12基因克隆及表达分析

为了探究谷胱甘肽转移酶编码基因(GST)在芥菜花青素积累中的作用,该文以紫薹-绿薹芥菜近等位基因系为材料,克隆到1个花青素积累相关的GST基因,命名为BjGSTF12。该文对BjGSTF12编码蛋白及其启动子进行生物信息学分析,并分析其在绿薹、紫薹芥菜中的表达水平及其与花青素含量的关系。结果表明:(1)BjGSTF12的基因组和cDNA全长分别为808、651 bp,编码216个氨基酸,具有GST_N端和GST_C端保守结构域。然而,绿薹、紫薹芥菜BjGSTF12序列无区别。(2)BjGSTF12与拟南芥AtGSTF12亲缘关系最近,同属于φ亚家族。(3)2个芥菜品系BjGSTF12启动子序列存在4处碱基突变/插入,但二者顺式作用元件种类与数目相同,均含9个MYB结合位点、1个赤霉素响应元件、3个非生物胁迫响应元件。(4)紫薹芥菜花青素含量显著高于绿薹芥菜,BjGSTF12表达水平与花青素含量表现出类似变化规律。(5)互作蛋白网络分析表明,BjGSTF12与花青素合成关键酶、糖基化修饰、转运蛋白等蛋白存在互作。综上认为,BjGSTF12在芥菜薹茎花青素积累中可能发挥重要作用,BjGSTF12可能通过互作蛋白调控芥菜花青素合成、修饰、转运从而影响花青素积累。该文对深入研究GST在芥菜薹茎花青素积累的功能及作用机制奠定了一定理论基础。

藜麦GST基因家族鉴定及冷胁迫下表达模式分析

冷胁迫会直接影响植物的生长和发育,并在植物体内不断累积有害物质。谷胱甘肽S-转移酶(glutathione S-transferase, GST)是一种对细胞保护至关重要的抗氧化酶,通过减少活性氧引起的生理性损伤,在生物和非生物应激反应中发挥重要作用。藜麦(Chenopodium quinoa Willd.)中富含蛋白质、氨基酸等对人体有益的物质,但藜麦苗期易受冷空气侵袭而导致减产。因此,鉴定藜麦耐寒相关基因是必要的,为此鉴定藜麦GST基因家族成员,并分析藜麦GST基因在冷胁迫下不同组织中的表达模式。结果表明,在藜麦基因组中共鉴定出59个藜麦GST基因成员,其氨基酸长度范围为199~416 aa,分子量在22.72~47.45 ku范围内,随机分布在14条染色体上,通过系统发育分析将其分为8个亚家族,在启动子区域中分析发现多个低温顺式作用元件(LTR)和多种生长发育及代谢相关的元件。在藜麦GST基因家族中共发现11对串联重复基因和11对片段重复基因,表明基因复制事件是该物种GST基因家族进化的主要驱动力。基因的组织特异性表达分析结果显示,大多数藜麦GST基因在不同持续时间的冷胁迫下可以作出响应,特别是在叶片上高表达。结果可为进一步研究藜麦GST基因功能提供基础,为藜麦耐寒品种选育提供候选基因。

Rapid discovery of a novel “green” and natural GST inhibitor for sensitizing hepatocellular carcinoma to Cisplatin by visual screening strategy

Over-expression of glutathione S-transferase(GST)can promote Cisplatin resistance in hepatocellular carcinoma(HCC)treatment.Hence,inhibiting GST is an attractive strategy to improve Cisplatin sensitivity in HCC therapy.Although several synthesized GST inhibitors have been developed,the side effects and narrow spectrum for anticancer seriously limit their clinical application.Considering the abundance of natural compounds with anticancer activity,this study developed a rapid fluorescence technique to screen“green”natural GST inhibitors with high specificity.The fluorescence assay demonstrated that schisanlactone B(hereafter abbreviated as C1)isolated from Xue tong significantly down-regulated GST levels in Cisplatin-resistant HCC cells in vitro and in vivo.Importantly,C1 can selectively kill HCC cells from normal liver cells,effectively improving the therapeutic effect of Cisplatin on HCC mice by downregulating GST expression.Considering the high GST levels in HCC patients,this compound demonstrated the high potential for sensitizing HCC therapy in clinical practice by down-regulating GST levels.

GST family genes in jujube actively respond to phytoplasma infection

Jujube witches’broom(JWB)caused by phytoplasma has a severely negative effect on multiple metabolisms in jujube.The GST gene family in plants participates in the regulation of a variety of biotic and abiotic stresses.This study aims to identify and reveal the changes in the jujube GST gene family in response to phytoplasma infection.Here,70 ZjGSTs were identified in the jujube genome and divided into 8 classes.Among them,the Tau-class,including 44 genes,was the largest.Phylogenetic analysis indicated that Tau-class genes were highly conserved among species,such as Arabidopsis,cotton,chickpea,and rice.Through chromosome location analysis,37.1%of genes were clustered,and 8 of 9 gene clusters were composed of Tau class members.Through RT-PCR,qRT-PCR and enzyme activity detection,the results showed that the expression of half(20/40)of the tested ZjGSTs was inhibited by phytoplasma infection in field and tissue culture conditions,and GST activity was also significantly reduced.In the resistant and susceptible varieties under phytoplasma infection,ZjGSTU49-ZjGSTU54 in the cluster IV showed opposite expression patterns,which may be due to functional divergence during evolution.Some upregulated genes(ZjGSTU45,ZjGSTU49,ZjGSTU59,and ZjGSTU70)might be involved in the process of jujube against JWB.The yeast two-hybrid results showed that all 6 Tauclass proteins tested could form homodimers or heterodimers.Overall,the comprehensive analysis of the jujube GST gene family revealed that ZjGSTs responded actively to phytoplasma infection.Furthermore,some screened genes(ZjGSTU24,ZjGSTU49-52,ZjGSTU70,and ZjDHAR10)will contribute to further functional studies of jujube-phytoplasma interactions.

Carboxylesterase and Glutathione-S-Transferase （GST＇s） Induced Resistance to Bacillus thuringiensis Toxin CrylAb in Rice Leaf Folder, Cnaphalocrocis medinalis （Guenee） Populations

The rice leaf folder （RLF）, Cnaphalocrocis medinalis （Guenee） （lnsecta： Lepidoptera： Pyralidae）, is an important pest, widely distributed in many rice growing areas of Asia. The over-use of broad-spectrum chemical insecticides has been cited as a major cause of outbreaks of C. medinalis as excessive spraying of insecticide disrupts natural biological control insecticides still remain the major control tactics against leaf folder. Carbofuran and fenthion, bendiocarb, acephate, carbosulfan, quinolphos, monocrotophos, phosphamidon and fenvalerate are the common ones used against rice leaf folder. Genetically, modified rice lines expressing B. thuringiensis insecticidal crystal proteins produced are highly tolerant to leidopteran pests. Though economic and environmental benefits of GM crops is well established, the matter of concern is the possibility of target insect pest developing resistance to this B. thuringiensis insecticidal toxins, evident from many laboratory and field experiments against many insect pests. The involvement of GSH S-transferase, carboxylesterase, and microsomal monooxygenase in insecticide resistance has been reported in insecticide-resistant strains of many insect species. Hence, the present study was taken up to monitor for cross resistance between B. thuringiensis cry toxins and synthetic insecticides in larvae of leaf folder as it is mediated by carboxylesterase titre and other enzymes by bioassay for two selected rice leaf folder field populations at the Entomology division of Directorate of Rice Research which showed 2-fold resistance ratio. Qualitative and quantitative changes of carboxylesterase （CarE） and glutathione-s-transferase （GST＇s） were worked out with midguts extracts of the two C. medinalis populations in the presence of a-napthyl acetate and chlorodi-nitro benzene substrates.

Modification of the Genetic Polymorphism of Glutathione-S-Transferase (GSTM1 and GSTT1) in Motorcycle Drivers Exposed to BTEX in Cotonou

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.

冷胁迫下甘蓝型冬油菜表达蛋白及BnGSTs基因家族的鉴定与分析

谷胱甘肽S-转移酶(glutathione S-transferases, GST)参与调节植物生长、发育和逆境胁迫反应的许多方面。本研究利用双向电泳和质谱技术分析‘16VHNTS309’在冷胁迫下差异表达的蛋白质,基于GO和KEGG分析鉴定出BE、APX、SOD、GST等参与冷胁迫反应的蛋白质。利用q RT-PCR和生理指标鉴定到响应冷胁迫的关键蛋白质GST,采用同源克隆法克隆到‘16VHNTS309’的GST基因。该基因CDS长度为642bp,编码213个氨基酸,是一个不稳定蛋白,属于GST_N_3谷胱甘肽S-转移酶家族,与甘蓝型油菜‘ZS11’序列相似性为99.22%,与‘ZS11’和‘Vision’两者的氨基酸序列相比较发现第127位亮氨酸(L)突变为脯氨酸(P)。基因家族分析表明,在甘蓝型油菜中共鉴定到153个Bn GSTs成员,按其功能主要分为Zeta、Phi、Theta、CHQ、DHAR、Lambda和Tau这七大类型,大部分的Bn GSTs属于Phi和Tau这2种类型。系统进化将Bn GSTs分为12个亚家族,亚家族Ⅰ和Ⅷ包含了较多的成员,Bn GSTs不均匀的分布在18条染色体上, C06染色体上分布Bn GSTs基因数量最多,含有10个保守的蛋白质基序。Bn GSTs基因家族中有99对基因存在共线性关系, 131个基因来自基因复制事件,片段重复事件在Bn GSTs基因的进化中起着重要作用。冷胁迫下Bna A02g35760D、Bna C06g20450D、Bna C06g35490D、Bna A02g03230D和Bna A02g35980D在强抗寒品种中的显著高表达,是弱抗寒品种的7~12倍,并且强抗寒品种具有较高的生理酶活性。另外,在冷冻胁迫下鉴定到一些瞬时和持续表达的关键候选基因。这些结果为进一步研究Bn GSTs基因在强抗寒甘蓝型冬油菜抗寒分子调控中的作用奠定基础。

食管胃交界部腺癌肿瘤浸润淋巴细胞中FoxM1、GST-π表达情况与分化程度、转移及预后的关系

目的 分析食管胃交界部腺癌(AEGJ)肿瘤浸润淋巴细胞(TILs)中叉头框蛋白M1(FoxM1)、谷胱甘肽-S-转移酶-Pi(GST-π)表达情况,探讨其与肿瘤分化程度、转移及患者预后之间的关系。方法 将60例行手术治疗的AEGJ患者纳为研究对象,术中留取癌灶组织与癌旁组织,机械法分离出癌灶组织及癌旁组织内TILs,流式技术检测TILs中FoxM1、GST-π表达量(以含有FoxM1、GST-π抗原细胞的百分率表示)。分析AEGJ癌灶组织及癌旁组织TILs中FoxM1、GST-π表达量与肿瘤临床特征之间的关系。随访3年,比较癌灶组织内TILs中FoxM1、GST-π不同表达量患者生存情况。结果 癌灶组织TILs内FoxM1、GST-π表达水平均高于癌旁组织TILs,差异具有统计学意义(P<0.05)。癌灶组织TILs内FoxM1表达水平与肿瘤TNM分期、浸润深度及淋巴转移相关(P<0.05),TILs内GST-π表达水平与肿瘤分化程度及浸润深度有关(P<0.05)。随访发现,FoxM1高表达组患者3年生存率及中位生存时间均低于低表达组(Rank=4.039,P=0.044),GST-π高表达组患者3年生存率及中位生存时间均低于低表达组(Rank=10.041,P=0.002)。结论 AEGJ组织TILs内FoxM1及GST-π表达水平明显高于癌旁TILs,且与AEGJ病理特点存在一定的关系。FoxM1及GST-π表达水平与AEGJ患者术后生存情况相关,是预测患者预后的潜在指标。

Pharmacogenetics of azathioprine in inflammatory bowel disease: A role for glutathione-S-transferase?

Azathioprine is a purine antimetabolite drug commonly used to treat inflammatory bowel disease (IBD). In vivo it is active after reaction with reduced glutathione (GSH) and conversion to mercaptopurine. Although this reaction may occur spontaneously, the presence of isoforms M and A of the enzyme glutathione-S-transferase (GST) may increase its speed. Indeed, in pediatric patients with IBD, deletion of GST-M1, which determines reduced enzymatic activity, was recently associated with reduced sensitivity to azathioprine and reduced production of azathioprine active metabolites. In addition to increase the activation of azathioprine to mercaptopurine, GSTs may contribute to azathioprine effects even by modulating GSH consumption, oxidative stress and apoptosis. Therefore, genetic polymorphisms in genes for GSTs may be useful to predict response to azathioprine even if more in vitro and clinical validation studies are needed.

Glutathione-S-transferase M1 polymorphisms on the susceptibility to esophageal cancer among three Chinese minorities:Kazakh,Tajik and Uygur

AIM： To investigate the glutathione-S-transferase M1 （GSTM1） polymorphisms in three Chinese minorities, Kazakh, Uygur, and Tajik; and the pathological significance of GSTM1 polymorphisms in esophageal carcinogenesis in Kazakh.METHODS： A total of 1121 blood samples （442 males and 679 females） were obtained from healthy Kazakh （654）, Uygur （412） and Tajik （55）. Primary esophageal squamous cell cancer （ESCC） tissues from Kazakh were obtained from 116 patients who underwent surgery. GSTM1 polymorphisms were analyzed by a combined approach of PCR and electrophoresis techniques.RESULTS： GSTM1 null genotype was found in 62.63% Uygur, 50.91% Tajik and 47.40% Kazakh. A significantly higher frequency of GSTM1 null genotype in Uygur was observed compared with Kazakh （OR： 1.859, 95% CI： 1.445 -2.391, χ^2 = 23.71, P = 0.000）. In addition, GSTM1 null genotype was found in 23.53% of welldifferentiated ESCC in Kazakh, in 49.23% of poorly differentiated ESCC, with a significant difference （OR： 3.152, 95% CI： 1.403-7.080, χ^2 = 8.018, P = 0.007）.CONCLUSION： There is a marked difference in the frequency of common GSTM1 null genotype between Uygur and Kazakh. GSTM1 null genotype is associated with differentiation of ESCC in Kazakh.

Glutathione-S-transferases genes-promising predictors of hepatic dysfunction

diseases pathogenesis are genes that encodes the synthesis of glutathione-Stransferase(GST),known as the second phase enzyme detoxification system that protects against endogenous oxidative stress and exogenous toxins,through catalisation of glutathione sulfuric groups conjugation and decontamination of lipid and deoxyribonucleic acid oxidation products.The group of GST enzymes consists of cytosolic,mitochondrial and microsomal fractions.Recently,eight classes of soluble cytoplasmic isoforms of GST enzymes are widely known:α-,ζ-,θ-,κ-,μ-,π-,σ-,andω-.The GSTs gene family in the Human Gene Nomenclature Committee,online database recorded over 20 functional genes.The level of GSTs expression is considered to be a crucial factor in determining the sensitivity of cells to a broad spectrum of toxins.Nevertheless,human GSTs genes have multiple and frequent polymorphisms that include the complete absence of the GSTM1 or the GSTT1 gene.Current review supports the position that genetic polymorphism of GST genes is involved in the pathogenesis of various liver diseases,particularly non-alcoholic fatty liver disease,hepatitis and liver cirrhosis of different etiology and hepatocellular carcinoma.Certain GST allelic variants were proven to be associated with susceptibility to hepatological pathology,and correlations with the natural course of the diseases were subsequently postulated.

β-catenin accumulation in nuclei of hepatocellular carcinoma cells up-regulates glutathione-s-transferase M3 mRNA

AIM: To identify the differentially over-expressed genes associated with β-catenin accumulation in nuclei of hepatocellular carcinoma (HCC) cells. METHODS: Differentially expressed genes were identified in radiation-induced B6C3 F1 mouse HCC cells by mRNA differential display, Northern blot and RT-PCR, respectively. Total glutathione-s-transferase (GST) activity was measured by GST activity assay and β-catenin localization was detected with immunostaining in radiation-induced mouse HCC cells and in HepG2 cell lines.RESULTS: Two up-regulated genes, glutamine synthetase and glutathione-s-transferase M3 (GSTM3), were identified in radiation-induced mouse HCC cells. Influence of β-catenin accumulation in nuclei of HCC cells on upregulation of GSTM3 mRNA was investigated. The nearby upstream domain of GSTM3 contained the β-catenin/TcfLef consensus binding site sequences [5'-(A/T)(A/T) CAAAG-3'], and the total GST activity ratio was considerably higher in B6C3F1 mouse HCC cells with β-catenin accumulation in nuclei of HCC cells than in those without β-catenin accumulation (0.353 ± 0.117vs 0.071 ± 0.064, P < 0.001). The TWS119 (a distinct GSK-3β inhibitor)-induced total GST activity was significantly higher in HepG2 cells with β-catenin accumulation than in those without β-catenin accumulation in nuclei of HCC cells. Additionally, the GSTM3 mRNA level was significantly higher at 24 h than at 12 h in TWS119-treated HepG2 cells. CONCLUSION: β-catenin accumulation increases GST activity in nuclei of HCC cells, and GSTM3 may be a novel target gene of the β-catenin/Tcf-Lef complex.

Prognostic Significance of Comparison of Clinical Indicators with Manifestations of Genetic Polymorphism of Glutathione-S-Transferases in Non-Small Cell Lung Cancer

The article presented the results of comparison of polymorphic variants of the genes GSTM1, GSTT1, GSTP1 and clinical manifestations of non-small cell lung carcinoma. The association of the genotype GSTT1 (del) with the risk of developing squamous cell lung cancer has been revealed (OR = 2.54 CI: 1.13 - 5.72, p = 0.035). Analysis of patient survival rate (n = 173) in groups of various histological types of lung cancer showed that in the group of squamous cell lung cancer (n = 91) in patients with genotype GSTT1 (del), the survival rate median was significantly higher—84 months (95% CI 12.4 - 155.7) than in patients with the genotype GSTT1 (+)—36 months (95% CI 25.2 - 46.8, p = 0.045). In contrast, in the adenocarcinoma group (n = 82), the survival rate median in patients with the genotype GSTT1 (del) was 19 months. (95% CI 6.2 - 33.5), and in patients with genotype GSTT1 (+)—67 months (95% CI 50.1 - 84.0), which is the basis for continuing this comparison in an additional group of testees, as the sampling did not achieve the reliability of p = 0.12. Hypothetically, these differences may be due to differences in the gender composition of squamous cell lung cancer and adenocarcinoma and the involvement of GST enzymes in the metabolism of estrogens in adenocarcinoma in women and other hormonal background and reactivity of the male body with squamous cell carcinoma. Further research and subsequent analysis of the results will be aimed at confirming this hypothesis.

Evaluation of Glutathione-S-transferase and ceruloplasmin levels in gingival crevicular fluid and gingival tissue as diagnostic markers for chronic periodontitis

Periodontitis, is an infectious ailment of multifactorial origin, that brings about destruction of bone and surrounding tissues. There are various oral pathogens that may be responsible for the destruction. The host encounters these microbial invasions and their products by the production and release of inflammatory mediators from the cells within the body. Glutathione-S-transferase (GST) are a group of enzymes that utilize glutathione in conditions resulting in oxidative stress. These enzymes play a key role in the detoxifycation of such substance. It aids in preventing damage to important cellular components caused by release of free reactive oxygen species. Ceruloplasmin is a ferroxidase enzyme. It plays a role as an anti-inflammatory agent, by its ability to scavenge free radicals within the body. The present study was targeted at evaluating the levels of Glutathione-S-Transferase (GST) and Ceruloplasmin as diagnostic markers for patients with chronic periodontitis in gingival crevicular fluid (GCF) and the gingival tissues. Thirty patients were divided into two groups. Experimental group comprising of 15 subjects with chronic perio- dontitis and the control group was composed of 15 healthy individuals. Highly significant changes in GST between the diseased and normal patients (P = 0.001) were detected. There was a decrease in GST level in both gingival tissue & GCF in diseased patients when compared to the control patients. The ceruloplasmin levels in GCF and gingival tissues showed no difference between the control and diseased group. Hence,these results indicate a relationship suggesting that GST produced during chronic inflammation could be used as biomarker that indicate periodontal disease .

响应槲皮素诱导的美国白蛾GST基因鉴定及表达分析

【目的】谷胱甘肽S-转移酶(GSTs)是昆虫体内一种重要的解毒酶,在植食性昆虫对植物次生物质的解毒适应中发挥重要作用。本研究筛选和鉴定了美国白蛾中肠响应槲皮素诱导的GST基因,分析了槲皮素对GST基因的诱导表达模式,为阐明美国白蛾GST基因在解毒代谢槲皮素中的功能奠定基础。【方法】基于槲皮素诱导的美国白蛾中肠转录组,筛选和鉴定响应槲皮素诱导的GST基因;通过构建系统发育树分析GST基因的家族类别;采用实时荧光定量PCR技术研究槲皮素对GST基因诱导的剂量效应和时间效应。【结果】鉴定了美国白蛾响应槲皮素诱导的6个GST基因;系统发育分析显示HcGST-E1、HcGST-E2、HcGST-E3属于GSTEpsilon家族,HcGST-S1、HcGST-S2属于GSTSigma家族,HcGST-O1属于GST Omega家族。不同质量分数槲皮素对GST基因的诱导作用不同,本实验质量分数(0.5%、1.0%、2.0%、4.0%)的槲皮素均能诱导HcGST-E1表达水平显著上调,0.5%、1.0%和2.0%的槲皮素诱导HcGST-E3表达水平显著升高;0.5%的槲皮素诱导HcGST-O1的表达水平显著增加。本实验质量分数的槲皮素均诱导HcGST-S1显著下调表达;0.5%和1.0%的槲皮素诱导HcGST-S2的表达水平显著下调。3个上调GST基因均在24 h或36 h内显著上调表达响应槲皮素的诱导。【结论】槲皮素能显著诱导美国白蛾中肠6个GST基因的表达水平,但对不同基因诱导的表达模式不同。HcGST-E1、HcGST-E3和HcGST-O1显著上调表达响应不同质量分数槲皮素的诱导,推测这3个基因是参与美国白蛾解毒代谢槲皮素的关键基因。

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

【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。

Immunoprophylactic potential of filarial glutathione-s-transferase in lymphatic filariaisis

Objective:To elucidates the immunoprophylactic potential of glutathion-s-transferase(GST) from cattle filarial parasite Setaria digitata(S.digitata) against lymphatic filariasis.Methods: GST was purified through affinity chromatography(SdGST) and chacterized by SDS-PAGE and Nano-LC MS/MS analysis.Antibody isotypes to SdGST were measured by ELISA.Antibody dependant cellular cytotoxicity(ADCC) was performed in vitro using sera from immunized animals and immune individuals.T-cell proliferation and cytokine response to SdGST in different groups of filariasis were measured.Immunoprophylactic potential of SdGST was evaluate in animal model.Results:SdGST exhibited 30-fold enhancement of enzyme activity over crude parasitic extract.It was found to be 26 kDa by SDS-PAGE.Nano LC-MS/MS analysis followed by blast search showed 100%homology with Dirqfilaria immitis(D.immitis) and only 43%with Homo sapiens(H.sapiens).Immunoblotting analysis showed putatively immune individuals carry significant level of antibodies to SdGST as compared with microfilaraemics.Immunized sera and sera endemic normal could neutralize the enzymatic activity of SdGST and inducing in vitro cytotoxicity of microfilariae.Peripheral blood mononuclear cells(PBMC) from endemic normals upon stimulation with SdGST showed a mixed type of Th1/Th2 response.SdGST immunization clear microfilariae from circulation in S.digitata implanted mastomys.Conclusions:The heterologous GST could be potentially developed as a vaccine candidate against lymphatic filarial parasite.

拟南芥AtGST8的功能及其过表达植株对甲基紫精胁迫的响应

谷胱甘肽S-转移酶(GST)可催化还原型谷胱甘肽(GSH)与底物连接,形成水溶性产物。植物受到胁迫时,GST会诱导表达,保护植物免受伤害,对植物抗逆起到重要作用。文章研究分析了拟南芥AtGST8的定位、表达及其过表达植株对甲基紫精(MV)胁迫的响应。结果表明:AtGST8定位在细胞膜。除种子和胚外,AtGST8在其余器官中均有表达,在莲座叶中表达较弱,而在花和果荚中的表达与生育进程有关。在果荚中表达量最高,叶中表达量最低。MV显著提高了所有供试材料幼苗中丙二醛(MDA)和抗坏血酸(AsA)含量,诱导超氧阴离子(O_(2)·^(-))的产生,过表达AtGST8植株的MDA和O_(2)·^(-)的增幅小于野生型,AsA则相反。MV处理明显降低了野生型中的还原型谷胱甘肽/氧化型谷胱甘肽(GSH/GSSG)水平,在过表达AtGST8植株中的变化则不明显;MV处理显著增加了所有供试材料中氧化胁迫保护酶(超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX))的活性;过表达AtGST8植株中GSH过氧化物酶(GSH-Px)和GST活性明显增大,而野生型变化不明显;MV处理的保护酶相关基因(SAPX、PRX34、CAT1、GST8)的表达水平与酶活表现一致。过表达AtGST8植株遭受MV胁迫后能够维持较高的AsA含量、GSH/GSSG水平、抗氧化酶活性及转录水平,从而及时清除体内的活性氧以减轻MV胁迫对植物的伤害。

益生菌结合蓝光照射治疗新生儿黄疸对体质量增加量及α-GST水平的效果分析

目的:探究益生菌结合蓝光照射治疗新生儿黄疸对体质量增加量及α-GST水平的影响。方法:研究对象选取我院2019年5月—2022年5月收治的黄疸新生儿80例,使用随机数字表法分为蓝光组及联合组,每组40例。蓝光组给予蓝光间歇性照射治疗,联合组在其基础上加用益生菌治疗。比价2组治疗疗效及治疗前后胆红素、营养状态、体质量增加量、α-GST、GLDH、ALT及神经损伤相关指标水平表达。结果:联合组治疗有效率显著高于蓝光组(P<0.05);治疗前2组血清胆红素无明显差异(P>0.05),治疗后,联合组血清胆红素水平显著低于蓝光组(P<0.05);治疗前,2组血清白蛋白、摄奶量差异无统计学意义(P>0.05),治疗后联合组2组血清白蛋白、母乳喂养吸吮次数、体质量增加量高于蓝光组(P<0.05);2组治疗前α-GST、GLDH、ALT水平差异无统计学意义(P>0.05),治疗后,联合组α-GST、GLDH、ALT低于蓝光组(P<0.05);治疗前,2组NBNA评分对比差异无统计学意义(P>0.05),治疗后联合组NBNA评分高于蓝光组,差异具有统计学意义(P<0.05)。结论:益生菌结合蓝光照射治疗新生儿黄疸有助于降低患儿机体胆红素水平,提高患儿机体营养状态及体质量,缓解患儿肝损伤及神经损伤症状,提高治疗效果。

Genetic polymorphism of glutathione-S-transferase M1 and T1 in associated with carcinogenesis of hepatocellular carcinoma and nasopharyngeal carcinoma

Objective:The aim of our study was to investigate the distribution of glutathione-S-transferase M1 (GSTM1) and T1 (GSTT1) gene polymorphism in hepatocellular carcinoma (HCC) and nasopharyngeal carcinoma (NPC) patients in a high risk area in Guangxi Zhuang Autonomous Region,China.Methods:It was a case-control study.The genotypes of GSTM1 and GSTT1 in 181 HCC and 126 NPC patients were compared with 641 matched control.The GSTM1 and GSTT1 genotypes were detected using conventional multiplex PCR method.Results:The frequency of GSTM1 null genotype in HCC,NPC and control groups were 65.2%,61.9% and 47.6% respectively,significant difference between these two cancer groups and control was observed (P < 0.01).The frequency of GSTT1 null genotype in HCC,NPC and control groups were 57.5%,62.7% and 43.1% respectively,significant difference between these two cancer groups and control was observed (P < 0.01).Conclusion:The distributions of GSTM1 and T1 genes are polymorphic in HCC and NPC patients in a high risk area in Guangxi,individuals with GSTM1-null or GSTT1-null would have an increasing risk of developing HCC and NPC,especially when combination with virus infection (HBV or EBV) and absorbed chemical toxin (AFB1 or cigarette).