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Transcriptome sequencing reveals neurotoxicity in embryonic neural stem/ progenitor cells under heat stress

TOXICOLOGY IN VITRO 2023年 86卷 105486页

作者： Wang, Zeze Luo, Xue Luo, Zhen Tan, Yulong He, Genlin Li, Ping Yang, Xuesen Army Med Univ Coll Mil Prevent Med Dept Trop Med 30th Gaotanyan Main St Chongqing 400038 Peoples R China Minist Educ China Key Lab Extreme Environm Med Chongqing Peoples R China

Neural tube malformation is a common kind of human birth defect. High temperature is one of the most common physical teratogenic factors. Several studies have suggested that heat stress may cause neurotoxicity during brain development, but more studies are warranted to reveal the mechanism and draw consistent conclusions. The current study used a cell model of primary mouse embryonic neural stem/progenitor cells (NSPCs) subjected to heat stress of 43 degrees C for 20 min. Our study investigated the changes in the NSPCs transcriptome under heat stress using high-throughput mRNA-seq. The NSPCs showed remarkably altered genes associated with cell growth, proliferation, cell cycle, and survival when exposed to heat stress. Heat stress reduced cell viability, proliferation, and neurosphere formation and caused cell cycle arrest and apoptosis in cultured NSPCs. pcr arrays confirmed that the TNF receptor family plays an important role in the apoptosis of NSPCs during heat stress. The results of real-time pcr confirmed that heat stress affects the expression of critical genes. We provide transcriptomic insight into heat stress-induced developmental neurotoxic effects and the underlying mechanisms.

关键词： Heat stress Neural stem progenitor cells Apoptosis Transcriptome pcr array

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Impact of chronic exposure of rats to bisphenol A from perinatal period to adulthood on intraprostatic levels of 5α-reductase isozymes, aromatase, and genes implicated in prostate cancer development

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ENVIRONMENTAL RESEARCH 2022年第PartA期212卷 113142页

作者： Sanchez, Pilar Castro, Beatriz Martinez-Rodriguez, Sergio Rios-Pelegrina, Rosa del Moral, Raimundo G. Torres, Jesus M. Ortega, Esperanza Univ Granada Sch Med Dept Biochem & Mol Biol Granada Spain Clin San Cecilio Univ Hosp Dept Anat Pathol Granada Spain Univ Granada Neurosci Inst Granada Spain

The synergetic effect of estrogens and androgens is known to play a crucial role in the physiopathology of the prostate gland. Bisphenol A (BPA) is an endocrine disrupting compound that can interfere with endocrine hormone functioning and thereby influence prostate development. The objective of this study was to examine the impact on prostate expression of aromatase, 5 alpha-R isozymes, and prostate cancer-related genes of exposure to low doses of BPA from perinatal period to adulthood. Vehicle or BPA (2.5 mu g/kg b.w./day) was administered to gestating Wistar rats from gestational day 12 (GD12) to parturition and then to their male pups from postnatal day 1 (PND1) until euthanization on PND90. Their prostate glands were examined by qRT-pcr, Western blot, pcr array, and morphological study. mRNA and protein levels of 5 alpha-R2 were significantly reduced and mRNA and protein levels of aromatase were significantly increased in BPA-treated animals, which also showed modifications of 8 out of the 84 key genes implicated in the development of prostate cancer. Because BPA interferes with genes involved in intraprostatic androgen and estrogen production and others implicated in prostate cancer, research is warranted into the prostate disease risk associated with chronic low-dose BPA exposure throughout life.

关键词： Bisphenol a 5 alpha-reductase isozymes Aromatase Prostate Prostate cancer pcr array

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NUP107在结直肠癌中的表达及生物学作用的研究

NUP107在结直肠癌中的表达及生物学作用的研究

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作者：黄佳茹吉林大学

学位级别：硕士

目的:本研究通过二维液相色谱-质谱联用技术来探究NUP107在结直肠癌(Colorectal cancer,CRC)及其相对应的癌旁组织中的表达量的差异,随后通过生物信息学,免疫印迹技术(Western blot)以及免疫组织化学技术(Immunohistochemistry)对质谱... 详细信息

目的: 本研究通过二维液相色谱-质谱联用技术来探究NUP107在结直肠癌(Colorectal cancer,CRC)及其相对应的癌旁组织中的表达量的差异,随后通过生物信息学,免疫印迹技术(Western blot)以及免疫组织化学技术(Immunohistochemistry)对质谱结果进行验证,最后利用体外的细胞功能实验来更深入地探究NUP107的表达量对结直肠癌细胞生物学功能的影响,为结直肠癌的诊断和治疗提供一个可能的新靶点和思路。方法: 1.利用液质联用技术来检测20对患者结直肠癌及其相对应的癌旁组织中的有关差异蛋白表达情况,查阅有关生信数据库,利用免疫印记技术验证NUP107在结直肠癌中的表达情况,并通过免疫组织化学技术对32例患者的病理切片标本进行分析来检测NUP107的蛋白表达情况。 2.实验选取两种结直肠癌目的细胞系,分别为HCT116和SW480,利用慢病毒感染构建NUP107敲低表达的细胞株,通过荧光显微镜下观察GFP的表达情况及RT-q pcr和Western Blot实验来验证NUP107稳转细胞株的感染效率,最后通过细胞克隆增殖,迁移,凋亡和细胞周期等实验来探究NUP107对结直肠癌细胞功能的影响。结果: 1.生信数据库分析,免疫印迹技术以及免疫组织化学技术均验证了NUP107在结直肠癌组织中上调表达。 2.成功构建了NUP107敲低细胞株,细胞增殖实验,迁移实验和凋亡实验等表明,NUP107敲低之后,细胞增殖能力和迁移能力明显减弱,凋亡明显增加。结论: ***107在结直肠癌细胞系中上调表达 2.敲低NUP107之后细胞增殖能力,迁移能力明显减弱,并促进细胞凋亡增加。 ***107可能作为结直肠癌诊断和治疗的新靶点发挥一定作用。

关键词：结直肠癌 NUP107 液质联用细胞周期 pcr array

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Prothioconazole and prothioconazole-desthio induced different hepatotoxicities via interfering with glycolipid metabolism in mice

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PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY 2022年 180卷 104983-104983页

作者： Tian, Sinuo Yan, Hang Meng, Zhiyuan Jia, Ming Sun, Wei Huang, Shiran Wang, Yu Zhou, Zhiqiang Diao, Jinling Zhu, Wentao China Agr Univ Coll Sci Innovat Ctr Pesticide Res Dept Appl Chem Yuanmingyuan West Rd 2 Beijing 100193 Peoples R China Hebei Univ Chinese Med Sch Basic Med Shijiazhuang 050200 Hebei Peoples R China Yangzhou Univ Sch Hort & Plant Protect Yangzhou 225009 Jiangsu Peoples R China

Prothioconazole (PTA), a new triazole fungicide, has been widely used worldwide. A recent study has confirmed that PTA and its main metabolite prothioconazole-desthio (dPTA) interfere with the liver metabolism in reptiles. However, little is known about liver toxicity of these two pollutants in mammals. Here, female mice were orally exposed to PTA (1.5 mg/kg body weight/day) and dPTA (1.5 mg/kg body weight/day) for 30 days. Additionally, growth phenotype and indexes related to serum and liver function were examined. Using metabolomics and gene expression analysis, PTA- and dPTA-induced hepatotoxicity was studied to clarify its potential underlying mechanism of action. Together, the results indicated that PTA and dPTA exposure caused changes in growth phenotypes, including elevated blood glucose levels, triglyceride accumulation, and damage of liver function. Additionally, exposure to PTA and dPTA caused changes in genes and metabolites related to glycolipid metabolism in female mice, thereby interfering with the pyruvate metabolism and glycolysis/gluconeogenesis pathways, ultimately leading to hepatic metabolism disorders. In particular, the effect of dPTA on hepatotoxicity has been proven to be more significant than that of PTA. Thus, these findings help us understand the underlying mechanism of action of PTA and dPTA exposure-induced hepatotoxicity in mammals and possibly humans.

关键词： Liver Metabolomic modulations pcr array Prothioconazole Prothioconazole-desthio

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Imazalil and its metabolite imazalil-M caused developmental toxicity in zebrafish (Danio rerio) embryos via cell apoptosis mediated by metabolic disorders

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PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY 2022年 184卷 105113页

作者： Huang, Shiran Huang, Ming Tian, Sinuo Meng, Zhiyuan Yan, Sen Teng, Miaomiao Zhou, Zhiqiang Diao, Jinling Zhu, Wentao China Agr Univ Coll Sci Innovat Ctr Pesticide Res Dept Appl Chem Beijing 100193 Peoples R China China Agr Univ Coll Sci Beijing 100193 Peoples R China Yangzhou Univ Sch Hort & Plant Protect Yangzhou 225009 Jiangsu Peoples R China Chinese Res Inst Environm Sci State Key Lab Environm Criteria & Risk Assessment Beijing 100012 Peoples R China Yuanmingyuan West Rd 2 Beijing 100193 Peoples R China

Imazalil (IMZ) is a highly effective fungicide employed in crop production. It has been consistently detected in aquatic environments. The main environmental metabolite of IMZ is imazalil-M (IMZ-M). Limited studies have focused on the toxicity of IMZ and IMZ-M in aquatic organisms. This study systematically evaluated the developmental toxicity of IMZ and IMZ-M on zebrafish (Danio rerio) embryos and explored the potential mechanisms involved. The results showed that IMZ and IMZ-M caused developmental toxicity, characterized by decreased heart rate, hatching inhibition, and pericardial cyst in zebrafish embryos. Subsequently, acridine orange (AO) staining revealed cell apoptosis in the area around the heart regions of zebrafish larvae. Besides, the expression levels of apoptosis-related genes also varied significantly. Furthermore, 1H NMR-based metabolomics analysis showed that IMZ and IMZ-M exposure could induce metabolic profiles disorder in zebrafish larvae. Importantly, zebrafish exposure to IMZ and IMZ-M significantly affected the metabolism of branched - chain amino acids, energy, and ketone bodies, which are related to cell apoptosis. Overall, the toxicity of IMZ and IMZM in zebrafish embryos and larvae was characterized, suggesting a theoretical basis for the potential environmental risks of IMZ and its metabolite IMZ-M on non-target organisms.

关键词： Zebrafish embryos Developmental toxicity Cell apoptosis Metabolomics pcr array

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MicroRNAs expression profiling in Egyptian colorectal cancer patients

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IUBMB LIFE 2020年第2期72卷 275-284页

作者： El Din, Noha G. Bader Ibrahim, Marwa K. El-Shenawy, Reem Satum, Ghada M. Farouk, Sally Zayed, Naglaa Khairy, Ahmed El Awady, Mostafa Natl Res Ctr Microbial Biotechnol Dept 33 EL Bohouth StPO 12622 Giza Egypt Cairo Univ Endem Med Dept Kasr Alainy Fac Med Giza Egypt

Egypt has increased incidence and high rate of early onset colorectal cancer (CRC). This study aimed to profile the expression levels of 84 circulating microRNAs (miRNAs) in Egyptian CRC patients and to evaluate the diagnostic accuracy of some selected miRNAs as diagnostic biomarkers for CRC patients. A total of 129 subjects (84 CRC patients and 45 healthy controls) were enrolled in two independent sample sets: the screening set (39 subjects) and the validation set (90 subjects). The expression profiles of 84 miRNAs were studied by miRNA pcr array in the screening set. Then four miRNAs (let-7c, 21, 26a, 146a) were selected to be studied by quantitative real-time pcr in the validation set. The pcr array results revealed significant up regulation of 20 miRNAs and downregulation of two miRNAs in CRC patients compared to the healthy subjects. Moreover, the expression levels of the four selected miRNAs were significantly higher in CRC serum samples than controls. The ROC analysis revealed that miRNAs (let-7c, 21, 26a and 146a) can effectively discriminate between CRC patients and the controls. The combination of the four miRNAs showed AUC of 0.950 (95% CI [0.898-1.002], p = .001). However, the combination of miR-21 and miR-26a showed the best diagnostic accuracy with AUC of 0.953 (95% CI [0.908-0.999], p = .001). The current data suggest that miRNAs (let-7c, 21, 26a, 146a) could play an important role in CRC development and they can be used as diagnostic biomarkers for CRC.

关键词： biomarkers colorectal cancer differential gene expression miRNAs pcr array

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叶酸对SOD1-G93A小鼠的促炎作用研究

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脑与神经疾病杂志 2024年第11期32卷 667-674页

作者：祝岩波严陶陶张双周巍河北省食品检验研究院石家庄050227

目的探讨叶酸对肌萎缩侧索硬化症(ALS)的可能作用机制,为叶酸在临床上的防治提供可靠的实验数据。方法将转基因SOD1-G93A突变型小鼠12对随机分为对照组和叶酸组,同窝分开,雌雄各半,每组12只。从60 d开始,对照组正常进水进食,叶酸组正常... 详细信息

目的探讨叶酸对肌萎缩侧索硬化症(ALS)的可能作用机制,为叶酸在临床上的防治提供可靠的实验数据。方法将转基因SOD1-G93A突变型小鼠12对随机分为对照组和叶酸组,同窝分开,雌雄各半,每组12只。从60 d开始,对照组正常进水进食,叶酸组正常进食,水添加4mg·kg^(-1)叶酸,每周称体质量2次。小鼠终末期脱颈处死后留取大脑、小脑、脊髓、心、肝、脾、肺、肾、坐骨神经、肌肉、小肠和结肠,提取RNA,pcr array试剂盒检测SOD1-G93A小鼠脊髓RNA表达水平,RT-qpcr检测SOD1-G93A小鼠大脑、小脑、脊髓、心、肝、脾、肺、肾、坐骨神经、肌肉、小肠和结肠中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、分化簇68(CD68)、分化簇86(CD86)和单核细胞趋化蛋白-1(MCP1)的表达水平。结果SOD1-G93A小鼠的叶酸组生存期为(128±8.1)d,对照组生存期为(136.5±9.7)d,叶酸组比对照组生存寿命平均减少8 d,差异有统计学意义(P<0.05);上调最为显著的基因有2个,分别为Epo和Bcl2a1a,涉及到Hypoxia和NF-κB信号通路。叶酸组在小脑和脊髓中TNF-α、IL-1β、CD68和CD 86的表达是对照组的两倍以上,叶酸组在小脑中MCP1的表达是对照组的两倍以上,差异有统计学意义(P<0.05)。叶酸组在心脏中TNF-α、IL-1β和MCP1的表达与对照组差异有统计学意义,在脾脏中IL-1β、CD68、CD86和MCP1的表达与对照组有显著性差异,其余无统计学意义。叶酸组在肌肉中TNF-α、CD68、CD86和MCP1的表达与对照组有显著性差异,叶酸组在结肠中TNF-α、IL-1β、CD68、CD86和MCP1的表达与对照组有显著性差异,其余差异无统计学意义。荧光染色结果表明SOD1-G93A小鼠脊髓中,Iba1和GFAP的表达叶酸组明显高于对照组。结论叶酸在SOD1-G93A小鼠模型中的干预可能激活了NF-κB信号通路,诱发了炎症反应,从而延长了病程。

关键词： pcr array 叶酸 SOD1-G93A小鼠炎症

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基于FXR-ASBT联级反应研究白头翁汤治疗UC的作用机制

基于FXR-ASBT联级反应研究白头翁汤治疗UC的作用机制

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作者：肖莹甘肃农业大学

学位级别：硕士

白头翁汤(Pulsatilla decoction,PD)是治疗溃疡性结肠炎的经典方剂,前期研究发现PD的治疗作用与法尼醇类受体(Farnesoid X receptor,FXR)有关,FXR活性受到顶端钠依赖性胆汁酸转运体(Apical sodium-dependent bile acid transporter,ASBT)的影响,而以胆汁酸受体FXR为核心的法尼醇X受体(FXR)-顶端钠依赖性胆汁酸转运体(ASBT)联级反应在维持胆汁酸(Bile acids,BAs)的代谢平衡对防止溃疡性结肠炎(ulcerative colitis,UC)的发生发展至关重要。因此,本论文采用pcr array筛选白头翁汤治疗溃疡性结肠炎胆汁酸通路差异基因,以从FXR-ASBT联级反应角度阐明PD治疗UC的作用机制。方法:将90只健康雄性BALB/C小鼠分空白对照组(CON)、模型组(DSS)、白头翁汤高剂量治疗组(PDHT,10.8g/kg)、白头翁汤中剂量治疗组(PDMT,5.4g/kg)、白头翁汤低剂量治疗组(PDLT,2.7g/kg)和柳氮磺胺嘧啶阳性对照组(SASP,0.45g/kg)。试验期间对各组小鼠临床症状进行疾病活动指数(Disease activity index,DAI)评分。糖原PAS、阿利新蓝染色检测结肠组织杯状细胞数量的情况。酶联免疫吸附法(Enzyme-linked immunosorbent assay,ELISA)检测血清和结肠组织中炎症因子的表达。pcr array芯片筛选白头翁汤治疗UC的88个与FXR-ASBT通路相关差异基因。蛋白质免疫印迹(Western blot)检测小鼠结肠组织筛选出差异基因的蛋白表达量。结果如下: 1、白头翁汤治疗后,小鼠疾病活动指数、结肠组织病理学损伤及结肠长度得到明显改善(P<0.01),结肠组织杯状细胞数量显著增加(P<0.01),血清和结肠组织中细胞因子IL-10显著升高(P<0.01);IL-23、IL-17、IL-1β显著降低(P<0.01),血清胆汁酸含量显著降低(P<0.01),胆固醇含量显著升高(P<0.01)。PD能缓解UC小鼠的症状,白头翁汤中剂量治疗组(PDMT)效果最佳。 2、pcr array结果显示,与CON组相比,DSS模型组小鼠结肠组织88个基因中共有42个基因表达显著变化,其中上调13个基因,下调29个基因。与DSS组相比,PDMT治疗恢复了:Nr1h4、Rxrα、IL17a、Cyp7a1、Abcb11、Cy p2a12、Fgf15、Slc27a5,Slc10a2、IL6、IL1b、Slco1a5、Slc51a、KIb、Slco1a1、Cyp27a1、Cyp8b1、Abcg8、Abcb1b、Cyp3a44、Abca基因。 3、WB结果显示,白头翁汤治疗后显著恢复了FXR、TGR5、CYP7A1、F GF15各蛋白的表达量(P<0.01),ABCB11和RXRα表达量有升高趋势,但无统计学意义。结论:白头翁汤对DSS诱导的UC小鼠具有良好的治疗作用。其作用机制为通过激活FXR-ASBT上下游基因,从而恢复紊乱的胆汁酸稳态、肠道屏障功能以及减轻肠道炎症有关。

关键词：白头翁汤 UC模型细胞因子 pcr array FXR ASBT

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New insights into bisphenols induced obesity in zebrafish (Danio rerio): Activation of cannabinoid receptor CB1

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JOURNAL OF HAZARDOUS MATERIALS 2021年 418卷 126100-126100页

作者： Tian, Sinuo Yan, Sen Meng, Zhiyuan Huang, Shiran Sun, Wei Jia, Ming Teng, Miaomiao Zhou, Zhiqiang Zhu, Wentao China Agr Univ Coll Sci Innovat Ctr Pesticide Res Dept Appl Chem Yuanmingyuan West Rd 2 Beijing 100193 Peoples R China Yangzhou Univ Sch Hort & Plant Protect Joint Int Res Lab Agr & Agriprod Safety Yangzhou 225009 Jiangsu Peoples R China Chinese Res Inst Environm Sci State Key Lab Environm Criteria & Risk Assessment Beijing 100012 Peoples R China

Bisphenols (BPs), as widely used plastic additives, penetrate into our daily lives. BPs are considered endocrine disruptors and could potentially induce obesity. In this study, the effects of bisphenol A (BPA) and tetrabromobisphenol A (TBBPA) on food intake and lipid metabolism in zebrafish were determined. Moreover, the impact of BPA and TBBPA on the endocannabinoid system (ECS) of zebrafish was further explored by metabolomics, transcriptomics, and molecular docking analysis. Here we show that exposure to BPA and TBBPA at concentrations commonly found in the environment (20, 100, and 500 mu g/L) led to hyperphagia and obesity in adult male zebrafish. Metabolomics and histopathological analysis revealed significant lipid accumulation in the liver of zebrafish exposed to BPA and TBBPA. The expression of ECS-related genes, in conjunction with RNA-Seq results, further indicated that BPA and TBBPA increased appetite and induced obesity by activating cannabinoid receptor type 1(CB1). Furthermore, molecular docking revealed that six representative BPs including BPA and TBBPA could bind to the CB1 receptor. Collectively, these findings indicate that CB1 may be a potential target for BPs to induce obesity.

关键词： Bisphenols Metabolomic modulations Molecular structure pcr array Zebrafish

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The Chondrogenic Potential of First-Trimester and Term Placental Mesenchymal Stem/Stromal Cells

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CARTILAGE 2021年第2_SUPPL期13卷 544S-558S页

作者： James, Joanna L. Umapathy, Anandita Srinivasan, Sonia Barker, Claire N. Brooks, Anna Hearn, James Chhana, Ashika Williams, Eloise Sheppard, Hilary McGlashan, Sue R. Univ Auckland Obstet & Gynaecol Auckland New Zealand Univ Auckland Anat & Med Imaging Auckland New Zealand Univ Auckland Sch Biol Sci Auckland New Zealand Univ Auckland Mol Med & Pathol Auckland New Zealand

Objectives Mesenchymal stem/stromal cells (MSCs) are a well-established cell source for cartilage engineering, but challenges remain as differentiation often results in chondrocyte hypertrophy. Chondrogenic potential also varies with MSC source and donor age. We assessed the chondrogenic potential of first-trimester and term placental MSCs and compared their response to commonly used bone marrow MSCs (BM-MSCs). Design MSCs were isolated from first-trimester and term placentae. BM-MSCs were commercially obtained. Chondrogenesis was induced by micromass culture in commercial chondrogenic media for 7, 14, or 21 days. Pellets were assessed for glycosaminoglycan (GAG) content, and types I, II, and X collagen. Gene expression was profiled using Qiagen RT2 human MSC arrays. Results At day 0, first-trimester and term MSCs expression levels of many chondrogenic genes to BM-MSC after 21 days of culture. Only first trimester MSCs showed significant changes in chondrogenic gene expression during induction compared to day 0 undifferentiated MSCs (greater BMP4, KAT2B, and reduced GDF6 expression). Additionally, first-trimester MSCs showed significantly greater expression of ABCB1 (at days 14 and 21) and BMP4 (at days 7, 14, 21) compared with term MSCs. Both first-trimester and term pellets showed increased GAG content over time and term MSCs had significantly GAG greater compared with BM-MSCs at days 7 and 14. Type II collagen was present in all pellets but unlike BM-MSCs, type I collagen was not observed in first-trimester or term MSC pellets. Conclusions These data highlight differences in BM-MSC and placental MSC chondrogenesis and demonstrate that placental MSCs may be an alternative cell source.

关键词： term placenta microarray chondrogenesis pcr array glycosaminoglycans

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