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.

Emerging trends and hotspots of Nuclear factor erythroid 2-related factor 2 in nervous system diseases

BACKGROUND The Nuclear factor erythroid 2-related factor 2(NRF2)transcription factor has attracted much attention in the context of neurological diseases.However,none of the studies have systematically clarified this field's research hotspots and evolution rules.AIM To investigate the research hotspots,evolution patterns,and future research trends in this field in recent years.METHODS We conducted a comprehensive literature search in the Web of Science Core Collection database using the following methods:(((((TS=(NFE2 L2))OR TS=(Nfe2 L2 protein,mouse))OR TS=(NF-E2-Related Factor 2))OR TS=(NRF2))OR TS=(NFE2L2))OR TS=(Nuclear factor erythroid2-related factor 2)AND(((((((TS=(neurological diseases))OR TS=(neurological disorder))OR TS=(brain disorder))OR TS=(brain injury))OR TS=(central nervous system disease))OR TS=(CNS disease))OR TS=(central nervous system disorder))OR TS=(CNS disorder)AND Language=English from 2010 to 2022.There are just two forms of literature available:Articles and reviews.Data were processed with the software Cite-Space(version 6.1.R6).RESULTS We analyzed 1884 articles from 200 schools in 72 countries/regions.Since 2015,the number of publications in this field has increased rapidly.China has the largest number of publications,but the articles published in the United States have better centrality and H-index.Among the top ten authors with the most published papers,five of them are from China,and the author with the most published papers is Wang Handong.The institution with the most articles was Nanjing University.To their credit,three of the top 10 most cited articles were written by Chinese scholars.The keyword co-occurrence map showed that"oxidative stress","NRF2","activation","expression"and"brain"were the five most frequently used keywords.CONCLUSION Research on the role of NRF2 in neurological diseases continues unabated.Researchers in developed countries published more influential papers,while Chinese scholars provided the largest number of articles.There have been numerous studies on the mechanism of NRF2 transcription factor in neurological diseases.NRF2 is also emerging as a potentially effective target for the treatment of neurological diseases.However,despite decades of research,our knowledge of NRF2 transcription factor in nervous system diseases is still limited.Further studies are needed in the future.

Focal lymphoblastic transformation of chronic myelogenous leukemia develops into erythroid leukemia:A case report

BACKGROUND We present a case of focal lymphoblastic transformation to erythroid leukemia following acute myeloblastic transformation in a patient with chronic myelogenous leukemia(CML)and discuss its mechanism of occurrence and development.CASE SUMMARY The presence of the Philadelphia(Ph)chromosome was identified through karyotype analysis,while the BCR-ABL fusion gene was detected using quantitative real-time polymerase chain reaction of the peripheral blood sample.Fluorescence in situ hybridization was used to detect the expression of the BCRABL gene in the lymphoma.Antigen expression and gene mutations in the primitive cells were detected by flow cytometry.The analysis confirmed the presence of CML along with focal lymphoblastic transformation to erythroid leukemia.Additionally,the patient was found to have secondary erythroid leukemia,along with multiple new gene mutations and abnormalities in complex karyotypes of chromosomes.CONCLUSION Our findings suggest a possible molecular basis for the focal lymphoblastic transformation secondary to myeloblastic transformation in patients with CML.

Neuroprotective effects of salidroside on focal cerebral ischemia/reperfusion injury involve the nuclear erythroid 2-related factor 2 pathway

Salidroside,the main active ingredient extracted from Rhodiola crenulata,has been shown to be neuroprotective in ischemic cerebral injury,but the underlying mechanism for this neuroprotection is poorly understood.In the current study,the neuroprotective effect of salidroside on cerebral ischemia-induced oxidative stress and the role of the nuclear factor erythroid 2-related factor 2（Nrf2）pathway was investigated in a rat model of middle cerebral artery occlusion.Salidroside（30 mg/kg）reduced infarct size,improved neurological function and histological changes,increased activity of superoxide dismutase and glutathione-S-transferase,and reduced malon-dialdehyde levels after cerebral ischemia and reperfusion.Furthermore,salidroside apparently increased Nrf2 and heme oxygenase-1 expression.These results suggest that salidroside exerts its neuroprotective effect against cerebral ischemia through anti-oxidant mechanisms and that activation of the Nrf2 pathway is involved.The Nrf2/antioxidant response element pathway may become a new therapeutic target for the treatment of ischemic stroke.

Is erythroferrone finally the long sought-after systemic erythroid regulator of iron?

Iron metabolism is regulated on the cellular and the systemic level. Over the last decade, the liver peptide "hepcidin" has emerged as the body's key irons store regulator. The long postulated "erythroid regulator of iron", however, remained elusive. Last year, evidence was provided, that a previously described myokine "myonectin" may also function as the long sought erythroid regulator of iron. Myonectin was therefore renamed "erythroferrone". This editorial provides a brief discussion on the two functions of erythroferrone and also briefly considers the emerging potential role of transferrin receptor 2 in erythropoiesis.

Short hairpin RNA-mediated knockdown of nuclear factor erythroid 2-like 3 exhibits tumor-suppressing effects in hepatocellular carcinoma cells

BACKGROUND Hepatocellular carcinoma(HCC) is one of the most common malignant tumors with high mortality-to-incidence ratios. Nuclear factor erythroid 2-like 3(NFE2 L3), also known as NRF3, is a member of the cap ‘n' collar basic-region leucine zipper family of transcription factors. NFE2 L3 is involved in the regulation of various biological processes, whereas its role in HCC has not been elucidated.AIM To explore the expression and biological function of NFE2 L3 in HCC.METHODS We analyzed the expression of NFE2 L3 in HCC tissues and its correlation with clinicopathological parameters based on The Cancer Genome Atlas(TCGA) data portal. Short hairpin RNA(shRNA) interference technology was utilized to knock down NFE2 L3 in vitro. Cell apoptosis, clone formation, proliferation, migration,and invasion assays were used to identify the biological effects of NFE2 L3 in BEL-7404 and SMMC-7721 cells. The expression of epithelial-mesenchymal transition(EMT) markers was examined by Western blot analysis.RESULTS TCGA analysis showed that NFE2 L3 expression was significantly positively correlated with tumor grade, T stage, and pathologic stage. The qPCR and Western blot results showed that both the mRNA and protein levels of NFE2 L3 were significantly decreased after shRNA-mediated knockdown in BEL-7404 and SMMC-7721 cells. The shRNA-mediated knockdown of NFE2 L3 could induce apoptosis and inhibit the clone formation and cell proliferation of SMMC-7721 and BEL-7404 cells. NFE2 L3 knockdown also significantly suppressed the migration, invasion, and EMT of the two cell lines.CONCLUSION Our study showed that shRNA-mediated knockdown of NFE2 L3 exhibited tumor-suppressing effects in HCC cells.

Effect on Proliferation and Erythroid Differentiation of K562 Cells by IER3IP1-Knockdown

Objective： To investigate the effect on erythroid differentiation and proliferation of K562 cells by IER3IP1-knockdown with RNA interference targeting at IER3IP1 gene. Methods： The shRNA eukaryotic expression vectors targeting at IER3IP1 gene were designed and constructed. Inhibitory effect was detected by semiquantitative RT-PCR. The impacts on K562 cells by RNAi were studied by MTT assay, benzidine staining, light microscope and electron microscopy observation, cell cycles analysis, colony formation assay and RT-PCR. The expressions of erythroid differentiation correlated genes Gfi-lB, GPA and 7-globin were studied after being exposed to 0.2μmol/L imatinib for two days. Results： The shRNA eukaryotic expression vectors were successfully constructed. The expression of IER3IP1 gene was significantly inhibited with an inhibition efficiency of 76% （P〈0.01）. Compared with the control groups, bcr/abl mRNA level was increased in K562/shRNA-IER3IP1 group （P〈0.01）. The proliferation ability was enhanced （P〈0.01） and the proportion of cells at G0/G1 phase decreased but S phase increased （P〈0.05） in K562/shRNA-IER3IP1 group. Under electron microscopy, the amount of euchromatin increased but heterochromatin decreased. There were structural abnomalities in endocytoplasmic reticulum and clusters of vesicular. The percentage of benzidine staining positive cells and mRNA expression levels of Gfi-1B, GPA and γ-globin were all decreased after being exposed to 0.2 μmol/L STI571 for two days in K562/shRNA-IER3IP1 group （P〈0.01）. Conclusion： IER3IP1-knockdown can hinder the erythroid differentiation and elevate the proliferation level of K562 cells. IER3IP1 may play a role in erythroid differentiation and proliferation of K562 cells.

Interplay between nuclear factor erythroid 2-related factor 2 and inflammatory mediators in COVID-19-related liver injury

Coronavirus disease 2019(COVID-19)caused by severe acute respiratory syndrome coronavirus 2 is a global pandemic and poses a major threat to human health worldwide.In addition to respiratory symptoms,COVID-19 is usually accompanied by systemic inflammation and liver damage in moderate and severe cases.Nuclear factor erythroid 2-related factor 2(NRF2)is a transcription factor that regulates the expression of antioxidant proteins,participating in COVID-19-mediated inflammation and liver injury.Here,we show the novel reciprocal regulation between NRF2 and inflammatory mediators associated with COVID-19-related liver injury.Additionally,we describe some mechanisms and treatment strategies.

原子因素的角色(导出 erythroid 2 ) 在新陈代谢的动态平衡和胰岛素行动喜欢 2 : 为糖尿病处理的一个新奇机会？

Redox balance is fundamentally important for physiological homeostasis. Pathological factors that disturb this dedicated balance may result in oxidative stress, leading to the development or aggravation of a variety of diseases, including diabetes mellitus, cardiovascular diseases, metabolic syndrome as well as in? ammation, aging and cancer. Thus, the capacity of endogenous free radical clearance can be of patho-physiological importance; in this regard, the major reactive oxy- gen species defense machinery, the nuclear factor (erythroid-derived 2)-like 2 (Nrf2) system needs to beprecisely modulated in response to pathological alterations. While oxidative stress is among the early events that lead to the development of insulin resistance, the activation of Nrf2 scavenging capacity leads to insulin sensitization. Furthermore, Nrf2 is evidently involved in regulating lipid metabolism. Here we summarize recent findings that link the Nrf2 system to metabolic homeostasis and insulin action and present our view that Nrf2 may serve as a novel drug target for diabetes and its complications.

Human adipose tissue contains erythroid progenitors expressing fetal hemoglobin

AIM: To investigate the origin of hematopoietic progenitors contained in the stromal vascular fraction(SVF)of human adipose tissue.METHODS: Tissue samples obtained from lipectomies were subjected to enzymatic digestion with collagenase to obtain a single-cell suspension. The centrifuged cell pellet, termed SVF, was separated immunomagnetically into CD45+and CD45-cells and cultured in serum-free medium containing hematopoietic cytokines. The freshly isolated and cultured cells were evaluated to determine their ability to form hematopoietic colony-forming units in clonogenic assays and for the expression of certain hematopoietic transcription factors by reversetranscription-polymerase chain reaction; the gene expression level was compared to that in CD34+hematopoietic progenitor cells from cord blood(CB) and adult peripheral blood(PB). To characterize erythroid progenitors, burst-forming units-erythroid(BFU-E) were developed in a semisolid medium under different culture conditions, and the hemoglobin composition and globin gene expression in the erythroid colonies were determined.RESULTS: The transcription factors SCL/TAL1, RUNX1,RUNX2 and GATA2 were expressed in both the CD45+and CD45-SVF populations; however, in contrast to our observations in the CD34+cells from CB and adult PB, GATA1 was not detected. Nevertheless, GATA1could be detected in the SVF cells after seven days in culture, whereas its expression was upregulated in the CB CD34+cells. The analysis of BFU-E-derived colonies revealed that virtually all erythroid cells produced by SVF cells expressed fetal hemoglobin, and the γ-globin mRNA levels ranged between those obtained in the adult- and neonatal-derived erythroid cells. Moreover,the SVF-derived erythroid cells synthesized similar levels of α- and β-globin mRNA, whereas the α-globin transcript levels were consistently higher those ofβ-globin in the cells derived from CB or PB CD34+cells. Furthermore, although the cellular distribution of hemoglobin in the erythroid cells derived from the CD34+cells obtained from hematopoietic tissues was dependent on the presence or absence of serum in the culture medium, this did not affect the SVF-derived erythroid cells.CONCLUSION: Our results demonstrate that hematopoietic progenitors in SVF have molecular and functional features that differ from those exhibited by circulating progenitors, suggesting the possibility of a different origin.

Nuclear factor erythroid 2-related factor 2-mediated signaling and metabolic associated fatty liver disease

Oxidative stress is a key driver in the development and progression of several diseases,including metabolic associated fatty liver disease(MAFLD).This condition includes a wide spectrum of pathological injuries,extending from simple steatosis to inflammation,fibrosis,cirrhosis,and hepatocellular carcinoma.Excessive buildup of lipids in the liver is strictly related to oxidative stress in MAFLD,progressing to liver fibrosis and cirrhosis.The nuclear factor erythroid 2-related factor 2(NRF2)is a master regulator of redox homeostasis.NRF2 plays an important role for cellular protection by inducing the expression of genes related to antioxidant,anti-inflammatory,and cytoprotective response.Consistent evidence demonstrates that NRF2 is involved in every step of MAFLD development,from simple steatosis to inflammation,advanced fibrosis,and initiation/progression of hepatocellular carcinoma.NRF2 activators regulate lipid metabolism and oxidative stress alleviating the fatty liver disease by inducing the expression of cytoprotective genes.Thus,modulating NRF2 activation is crucial not only in understanding specific mechanisms underlying MAFLD progression but also to characterize effective therapeutic strategies.This review outlined the current knowledge on the effects of NRF2 pathway,modulators,and mechanisms involved in the therapeutic implications of liver steatosis,inflammation,and fibrosis in MAFLD.

Downregulation of the Spi-1/PU.1 oncogene induces the expression of TRIM10/HERF1, a key factor required for terminal erythroid cell differentiation and survival

Spi-1/PU.1 和 Fli-1 oncoproteins 的持续表示在老鼠 erythroleukemia 房间堵住 globin 基因激活；然而，仅仅 Spi-1/PU.1 表示禁止 exon 的包括 16 在成熟 4.1R mRNA。这个拼接的事件为红血房间膜正直为功能的 4.1R 蛋白质并且，因此是关键的。这份报告证明 Spi-1/PU.1 downregulation 导致 TRIM10/hematopoietic 戒指手指 1 的激活(HERF1 ) ，分成三部分的主题(修剪) 的一个成员为 globin 基因抄写需要的 /RBCC 蛋白质家庭。另外，我们证明 TRIM10/HERF1 为 exon 拼接调整被要求 16 在迟了的 erythroid 区别期间。用可诱导的 overexpression 和 silencing 途径，我们发现了那：(1 ) TRIM10/HERF1 击倒在导致的 dimethylsulfoxide (DMSO ) 禁止拼接的血红素生产和 exon 和扳机房间 apoptosis 房间；(2 ) TRIM10/HERF1 upregulation 被要求，但是在它的自己上是不够的激活 exon 保留；(3 ) Fli-1 没在 TRIM10/HERF1 表示上有效果，而也导致 DMSO 的 downregulation 或 Spi-1/PU.1 shRNA 击倒表示是足够的激活 TRIM10/HERF1 表示；并且(4 ) Spi-1/PU.1 击倒的扳机抄写和拼接的事件独立于化学正式就职。总的来说，这些数据显示主要 Spi-1/PU.1 downregulation 通过至少二条小径，其一条要求 TRIM10/HERF1 upregulation 和平行对迟了的 erythroid 区别起作用 Spi-1/PU.1-induced Fli-1 用以遮闭之物规章的串联。

A critical role for the co-repressor N-CoR in erythroid differentiation and heme synthesis

合作抑压者 N 英国管(原子受体合作抑压者) 有重要角色冷淡的生物学过程包括增长，区别和开发。Mutantmice 缺乏 N 英国管是胚胎的联盟者 lethal 并且显得由于缺点从贫血症死非决定性红血球生成。然而， N-CoR-mediated erythroiddifferentiation 的内在的分子的机制大部分是未知的。化学上使用人的 erythroleukemic K562 房间线，它能是导致了系看情况区分进 erythroid 或 megakaryocytic 在 inducers 使用了，我们在 erythroid 区别调查了 N 英国管的角色。我们显示出那 N 英国管任何一个击倒短暂地(siRNA ) 或(shRNA ) 永久地损害 cytosinearabinoside (Ara-C )- 然而并非 K562 房间的导致 hemin 的 erythroid 区别。RT-PCR 分析表明 N 英国管被 5-aminolevulinate synthase (ALA-S2 ) 的 Ara-C 为正式就职要求，涉及的关键酶他我生合成。而且， N 英国管蛋白质的数量在 Ara-C-induced K562 区别期间显著地增加，显然通过 post-transcriptionalmechanism。与从 N-CoR-null 鼠标的数据一致， N 英国管没为 K562 房间的区别被要求进 megakaryocytic 系，由 phorbol 12-myristate13-acetate 导致了。因此，我们的试管内学习在 erythroid 区别为 N 英国管证实一个角色并且第一次表明 N 英国管为涉及 hemesynthesis 的关键酶的正式就职被要求。

DIFFERENTIATION AND MALIGNANT SUPPRESSION INDUCED BY MOUSE ERYTHROID DIFFERENTIATION AND DENUCLEATION FACTOR ON MOUSE ERYTHROLEUKEMIA CELLS

Objective.To investigate the roles of mouse erythroid differentiation and denucleation factor(MEDDF),a novel factor cloned in our laboratory recently ,in erythroid terminal differentiation.Methods.Mouse erythroleukemia(MEL) cells were transfected with eukaryotic expression plasmid pcDNA-MEDDF.Then we investigated the changes on characteristics of cell growth by analyzing cells growth rate,mitotic index and colony-forming rate in semi-soid medium.The expressions of c-myc and β-globin genes were analysed by semi-quantitative RT-PCR。Results.MEL cells transfected with pcDNA-MEDDF showed significant lower growth rate,mitotic index,and colony-forming rate in semi-solid medium(P<0.01).The percentage of benzidine-positive cells was 32.8% after transfection.The expression of β-globin in cells transfected with pcDNA-MEDDF was 3.43 times higher than that of control(MEL transfected with blank vector,pcDNA3.1) ,and the expression of c-myc decreased by 66.3%.Conclusions.MEDDF can induce differentiation of MEL cell and suppress its malignancy.

Identification of key genes responsible for cytokine-induced erythroid and myeloid differentiation and switching of hematopoietic stem cells by RAGE

我们利用了唯一的耕灼度分析基因，蛋白质，和房间表面的表示模式相关基因 inerythroid 和 myeloid 区别的抗原，和生物学过程并且响应 cytokine 改变造血的干细胞(HSC ) 切换。基因特定的碎片(266 ) 从五张人口鉴别刺激 ofcytokine 的 HSC 被分成三个组：(1 ) 在一张单个房间人口明确地表示了；(2 ) 在二张房间人口表示了，并且(3 ) 在三表示了或更多的人口。145 定义 cDNAs，(2%) 三是新奇基因。显示出的蛋白质二维的胶化电气泳动和流动 cytometry 分析重叠了并且在人口学习了的房间区分了蛋白质表示侧面。生物学过程印射在 erythroidand myeloid 系表示的 mRNAs 显示 mRNAs 由出席的两个系分享了“核心过程，“而明确地独自在任何一个系表示的基因与特定的过程或细胞的成熟有关。从这研究的数据支持承诺了 HSC (E14 或 G14 ) 的假设当 erythropoietin (EPO ) 被分别地在 myeloid 有教养的环境与 EPO 在 erythroid 有教养的条件 orG-CSF 下面刺激因素(G-CSF ) 的 granulocyte 殖民地代替时，房间能仍然被重定向发展成 myeloid 或 erythroid 房间。我们的结果建议基因或蛋白质在 erythroid 共同表示， myeloid 系可能为系维护是必要的并且在造血作用切换。

lncRNA NEAT1 is required for splenic erythroid differentiation

The erythropoiesis process consists of differentiation stages,starting with erythroid progenitors,including erythroid burstforming unit cells(BFU-Es),erythroid colony-forming unit cells(CFU-Es),followed by terminal erythroid differentiation,which,in turn,gives rise to basophilic erythroblasts,polychromatic erythroblasts,orthochromatic erythroblasts,reticulocytes(RETs),and mature erythrocytes(Hu et al.,2013).

Ultrastructural characteristics of erythroid cells in congenital dyserythropoietic anemia typeⅡ,with a focus on peripheral cisternae and double membranes

Peripheral cisternae and double membranes(PCDMs)in erythroid cells are a landmark of typeⅡcongenital dyserythropoietic anemia(CDA).To gain further insights into the mechanism of dyserythropoiesis,erythroblasts and erythrocytes in bone marrow were studied in 22 Chinese patients with CDAⅡby transmission electron microscopy.The study demonstrated an increase in all patients in erythroblasts with PCDMs with development from pro-erythroblast to red blood cells.PCDMs often connected with cisternae of endoplasmic reticulum(ER)and the perinuclear space,and were accompanied by karyopyknosis,karyolysis and disruption in polychromatic and orthochromatic erythroblasts.The results suggest that PCDMs are transformed from ER during erythropoiesis and participate in the dissolution and deletion of late erythroid cells in patients with CDAⅡ.

岩藻黄质活化核因子E2相关因子2改善糖皮质激素诱导的成骨细胞凋亡

背景:骨质疏松症发病率高,易导致骨折等并发症的发生,而现有药物干预不良反应大,难以满足临床需求。目的:探索岩藻黄质对糖皮质激素诱导成骨细胞骨质疏松症模型的作用与潜在机制。方法:将原代大鼠成骨细胞接种于6孔板内,待细胞融合度达到80%后分4组干预:对照组单纯培养24 h,糖皮质激素组使用地塞米松干预24 h,岩藻黄质组使用岩藻黄质干预24 h,糖皮质激素+岩藻黄质组使用地塞米松与岩藻黄质同时干预24 h。干预结束后,检测细胞增殖、凋亡、细胞内活性氧含量以及凋亡相关蛋白、骨形成相关蛋白、细胞核核因子E2相关因子2的蛋白表达。结果与结论:①CCK-8检测显示,与对照组比较,糖皮质激素组细胞活性降低(P<0.05);与糖皮质激素组比较,糖皮质激素+岩藻黄质组细胞活性升高(P<0.05);②JC-1线粒体膜电位染色与流式细胞学检测显示,与对照组比较,糖皮质激素组细胞凋亡比例增加(P<0.05);与糖皮质激素组比较,糖皮质激素+岩藻黄质组细胞凋亡比例减少(P<0.05);③Western Blot检测显示,与对照组比较,糖皮质激素组BAX、裂解聚ADP核糖聚合酶的蛋白表达升高(P<0.05),BCL2、Ⅰ型胶原蛋白α1肽链、碱性磷酸酶、骨钙蛋白、RUNX2的蛋白表达降低(P<0.05);与糖皮质激素组比较,糖皮质激素+岩藻黄质组BAX、裂解聚ADP核糖聚合酶的蛋白表达降低(P<0.05),BCL2、Ⅰ型胶原蛋白α1肽链、碱性磷酸酶、骨钙蛋白、RUNX2的蛋白表达升高(P<0.05);④荧光探针检测显示,与对照组比较,糖皮质激素组活性氧含量增加(P<0.05);与糖皮质激素组比较,糖皮质激素+岩藻黄质组活性氧含量减少(P<0.05);⑤免疫荧光染色与Western Blot检测显示,与对照组比较,糖皮质激素组细胞核核因子E2相关因子2的蛋白表达降低(P<0.05);与糖皮质激素组比较,糖皮质激素+岩藻黄质组细胞核核因子E2相关因子2的蛋白表达升高(P<0.05);⑥结果表明,岩藻黄质通过活化核因子E2相关因子2改善糖皮质激素诱导的成骨细胞凋亡与骨形成相关分子表达。

金合欢素调节Sirt1/AMPK/Nrf2信号通路对糖尿病白内障大鼠氧化应激损伤的影响

目的探讨金合欢素对糖尿病白内障(DC)大鼠氧化应激损伤的影响及其对沉默调节蛋白1(Sirt1)/腺苷酸活化蛋白激酶(AMPK)/核因子E2相关因子2(Nrf2)信号通路的调控作用。方法60只SD大鼠随机分为对照组、模型组、金合欢素低剂量组、金合欢素高剂量组、金合欢素+Sirt1抑制剂(EX527)组,除对照组以外均构建DC大鼠模型,其中,金合欢素低剂量组、金合欢素高剂量组大鼠分别经颈部皮下注射10 mg·kg^(-1)、20 mg·kg^(-1)的金合欢素,金合欢素+EX527组大鼠经颈部皮下注射20 mg·kg^(-1)金合欢素,均为每天2次,同时金合欢素+EX527组大鼠经皮下埋入渗透微型泵每天泵入3.5 mg·kg^(-1)EX527,其余组别均泵入等量生理盐水,给药持续4周。给药结束后,测量血压和空腹血糖(FBG),裂隙灯照射法观察大鼠晶状体混浊状况,HE染色观察晶状体组织病理学变化,ELISA测定血清丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、白细胞介素(IL)-6、IL-1β的含量,Western blot检测Sirt1、p-AMPK、AMPK、Nrf2蛋白表达水平。结果与对照组相比,模型组大鼠晶状体上皮细胞呈片状、条索状,发生迁移性聚集,收缩压、FBG、晶状体混浊评分、MDA、IL-6、IL-1β水平均升高,SOD、GSH-Px含量及Sirt1、p-AMPK/AMPK、Nrf2蛋白表达水平均降低(均为P<0.05);与模型组比较,金合欢素低、高剂量组大鼠晶状体上皮细胞迁移性聚集现象改善,收缩压、FBG、晶状体混浊评分、MDA、IL-6、IL-1β水平均降低,SOD、GSH-Px含量及Sirt1、p-AMPK/AMPK、Nrf2蛋白表达水平均升高(均为P<0.05);与金合欢素高剂量组比较,金合欢素+EX527组晶状体上皮细胞形态改变和聚集现象加重,收缩压、FBG、晶状体混浊评分、MDA、IL-6、IL-1β水平均升高,SOD、GSH-Px含量及Sirt1、p-AMPK/AMPK、Nrf2蛋白表达水平均降低(均为P<0.05)。结论金合欢素可能通过激活Sirt1/AMPK/Nrf2通路保护DC大鼠免受氧化应激损伤。

姜酮通过激活Nrf2/HO-1信号通路减轻OGD/R后氧化应激损伤对HT22细胞凋亡的抑制作用

目的:探讨姜酮对氧糖剥夺/复糖复氧(OGD/R)后小鼠海马神经元HT22细胞的保护作用,阐明其相关作用机制。方法:培养HT22细胞,设置不同OGD/R时间梯度,建立OGD/R细胞损伤模型。HT22细胞分为对照组、OGD/R组、OGD/R+1μmol·L^(-1)姜酮组、OGD/R+10μmol·L^(-1)姜酮、OGD/R+100μmol·L^(-1)姜酮组和OGD/R+0.2%二甲亚枫(DMSO)组,CCK-8法检测各组细胞活性并计算各组细胞存活率,确定姜酮最适药物浓度。细胞分为对照组、OGD/R组、OGD/R+姜酮组和OGD/R+姜酮+核因子E2相关因子2(Nrf2)抑制剂(ML385)组,OGD/R+姜酮组细胞经姜酮给药处理4 h后予以OGD 8 h和复糖复氧8 h处理,OGD/R+姜酮+ML385组细胞在姜酮给药前予以10μmol·L^(-1)ML385预处理6 h,CCK-8法检测各组细胞活性,Western blotting法检测各组细胞中Nrf2、血红素加氧酶1(HO-1)、B细胞淋巴瘤2(Bcl-2)和Bcl-2相关X蛋白(Bax)蛋白表达水平,酶联免疫吸附试验(ELISA)法检测各组细胞培养上清中超氧化物歧化酶(SOD)活性和丙二醛(MDA)水平。结果:与对照组比较,HT22细胞经OGD 8 h和复糖复糖8 h处理后细胞存活率低于50%,以OGD 8 h和复糖复糖8 h建立HT22细胞OGD/R模型。与OGD/R组比较,OGD/R+不同剂量姜酮组细胞存活率均不同程度升高,其中OGD/R+100μmol·L^(-1)姜酮组细胞存活率升高最明显(P<0.01),故选用100μmol·L^(-1)姜酮用于后续实验。与对照组比较,OGD/R组细胞活性明显降低(P<0.01),细胞中Nrf2、HO-1和Bax蛋白表达水平明显升高(P<0.01),Bcl-2蛋白表达水平明显降低(P<0.05),细胞培养上清中SOD活性明显降低(P<0.01),MDA水平明显升高(P<0.01);与OGD/R组比较,OGD/R+姜酮组细胞活性明显升高(P<0.01),细胞中Nrf2、HO-1和Bcl-2蛋白表达水平明显升高(P<0.05或P<0.01),Bax蛋白表达水平明显降低(P<0.05),细胞培养上清中SOD活性明显升高(P<0.01),MDA水平明显降低(P<0.01);与OGD/R+姜酮组比较,OGD/R+姜酮+ML385组细胞活性明显降低(P<0.01),细胞中Nrf2、HO-1和Bcl-2蛋白表达水平明显降低(P<0.01),Bax蛋白表达水平明显升高(P<0.01),细胞培养上清中SOD活性明显降低(P<0.01),MDA水平明显升高(P<0.05)。结论:姜酮可通过激活Nrf2/HO-1信号通路减轻OGD/R后氧化应激损伤对HT22细胞凋亡的抑制作用。