Neferine inhibits the progression of diabetic nephropathy by modulating the miR-17-5p/nuclear factor E2-related factor 2 axis

OBJECTIVE:To investigate the effect of Neferine(Nef)on diabetic nephropathy(DN)and to explore the mechanism of Nef in DN based on miRNA regulation theory.METHODS:A DN mouse model was constructed and treated with Nef.Serum creatinine(Crea),blood urea(UREA)and urinary albumin were measured in mice by kits,and renal histopathological changes and fibrosis were observed by hematoxylin-eosin staining and Masson staining.Renal tissue superoxide dismutase(SOD),malondialdehyde(MDA)and glutathione peroxidase(GSH-Px)activities were measured by enzyme-linked immunosorbent assay(ELISA).Western blotting was used to detect the expression of nuclear factor E2-related factor 2(Nrf2)/heme oxygenase 1(HO-1)signaling pathway-related proteins in kidney tissues.Quantitative reverse transcription-polymerase chain reaction(q RT-PCR)was used to detect the expression of miR-17-5p in kidney tissues.Subsequently,a DN in vitro model was constructed by high glucose culture of human mesangial cells(HMCs),cells were transfected with miR-17-5p mimic and/or treated with Nef,and we used q RTPCR to detect cellular miR-17 expression,flow cytometry to detect apoptosis,ELISAs to detect cellular SOD,MDA,and GSH-Px activities,Western blots to detect Nrf2/HO-1 signaling pathway-related protein expression,and dual luciferase reporter gene assays to verify the targeting relationship between Nrf2 and miR-17-5p.RESULTS:Administration of Nef significantly reduced the levels of blood glucose,Crea,and UREA and the expression of miR-17-5p,improved renal histopathology and fibrosis,significantly reduced MDA levels,elevated SOD and GSH-Px activities,and activated Nrf2 expression in kidney tissues from mice with DN.Nrf2 is a post-transcriptional target of miR-17-5p.In HMCs transfected with miR-17-5p mimics,the m RNA and protein levels of Nrf2 were significantly suppressed.Furthermore,miR-17-5p overexpression and Nef intervention resulted in a significant increase in high glucose-induced apoptosis and MDA levels in HMCs and a significant decrease in the protein expression of HO-1 and Nrf2.CONCLUSION:Collectively,these results indicate that Nef has an ameliorative effect on DN,and the mechanism may be through the miR-17-5p/Nrf2 pathway.

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.

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.

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.

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.

Keap1-nuclear factor rythroid 2-related factor 2 inhibitor NXPZ ameliorates Aβ1-42-induced cognitive dysfunction in mice

OBJECTIVE Nuclear factor erythroid 2-related factor 2(Nrf2) is found to be ubiquitiously expressed in many tissues,and works as the key regulator against oxidative stress damage in cells and organs,which makes Nrf2 a widely concerned drug target.Recent research has identified that Nrf2 is involved in the pathology of Alzheimer disease(AD),whereas the mechanism is unknown.The purpose of this study is to figure out the role of Nrf2 in the pathologic process of AD through Nrf2-Keap1-ARE pathway and the effects of Keap1-Nrf2 inhibitor in AD mice models.METHODS Amyloid β^(1-42)(Aβ^(1-42))was injected into the bilateral hippocampus to induce the cognitive dysfunction in eight-week old male mice.The mice were treated with Keap1-Nrf2 inhibitor NXPZ of three doses as well as donepezil as a positive control by intragastric administration one time a day for one week.Several behavior tests were used to analyze the mice learning and memory ability.Additionally,we detected Nrf2 and Aβ in the plasma in mice with ELISA kits,as well as some factors related to oxidative stress in the hippocampus and cortex.The expression levels of Nrf2,Keap1,Tau and p-Tau were measured in the murine brain tissue with Western blotting.SH-SY5 Y cells were studied as an in vitro model to further clarify the mechanism.RESULTS The treatment of NXPZ ameliorated learning and memory dysfunction in AD mice in a dose-dependent manner,and the high dose group recovered better than the positive drug group.The plasma Nrf2 level was increased in a dose-dependent manner in the treatment groups;however,the plasma Aβ was decreased.What′ s more,superoxide dismutase(SOD) and glutathione reductase(GSSH) in the hippocampus and cortex were increased in the treatment group,while the malondialdehyde(MDA) was decreased,meaning that NXPZ treatment promoted expression of the anti-oxidative factors and inhibited the expression of the oxidative factors in the down-stream.Western blotting analysis of hippocampus and cortex showed up-regulated Nrf2,decreased Keap1 and decreased p-Tau in NXPZ treatment mice.In ex vivo experiments,when SH-SY5 Y cells were treated with Aβ,Nrf2 in the cytoplasm was increased,as well as the expression Nrf2 in the nuclear was decreased.The treatment of NXPZ increased nuclear Nrf2,decreased cytoplasm Nrf2,and decreased the expression of p-Tau.CONCLUSION Nrf2 has an important role in neuron function.Nrf2 activation by selective Keap1-Nrf2 inhibitor NXPZ may contribute to improve cognitive function in AD mice.The mechanism may be related to increased generation and release of Nrf2 induced by more disaggregation with Keap1,leading to more expression of anti-oxidative molecules to protect the damage caused by Aβ.These results indicates that Nrf2 may be a novel therapeutic target of AD and Keap1-Nrf2 inhibitor may be a novel medication for protecting the loss of learning and memory ability.

Long non-coding RNA CDKN2B-AS1 promotes hepatocellular carcinoma progression via E2F transcription factor 1/G protein subunit alpha Z axis

BACKGROUND A series of long non-coding RNAs(lncRNAs)have been reported to play a crucial role in cancer biology.Some previous studies report that lncRNA CDKN2B-AS1 is involved in some human malignancies.However,its role in hepatocellular carcinoma(HCC)has not been fully deciphered.AIM To decipher the role of CDKN2B-AS1 in the progression of HCC.METHODS CDKN2B-AS1 expression in HCC was detected by quantitative real-time polymerase chain reaction.The malignant phenotypes of Li-7 and SNU-182 cells were detected by the CCK-8 method,EdU method,and flow cytometry,respectively.RNA immunoprecipitation was executed to confirm the interaction between CDKN2B-AS1 and E2F transcription factor 1(E2F1).Luciferase reporter assay and chromatin immunoprecipitation were performed to verify the binding of E2F1 to the promoter of G protein subunit alpha Z(GNAZ).E2F1 and GNAZ were detected by western blot in HCC cells.RESULTS In HCC tissues,CDKN2B-AS1 was upregulated.Depletion of CDKN2B-AS1 inhibited the proliferation of HCC cells,and the depletion of CDKN2B-AS1 also induced cell cycle arrest and apoptosis.CDKN2B-AS1 could interact with E2F1.Depletion of CDKN2B-AS1 inhibited the binding of E2F1 to the GNAZ promoter region.Overexpression of E2F1 reversed the biological effects of depletion of CDKN2B-AS1 on the malignant behaviors of HCC cells.CONCLUSION CDKN2B-AS1 recruits E2F1 to facilitate GNAZ transcription to promote HCC progression.

Keap1/Nrf2信号通路在非小细胞肺癌氧化应激机制中的作用

目的检测非小细胞肺癌(NSCLC)组织中Kelch样环氧氯丙烷相关蛋白-1(Keap1)、核因子E2相关因子2(Nrf2)蛋白表达水平,分析其与临床病理参数、氧化应激指标的相关性,为临床治疗提供潜在靶点。方法选取2017年4月至2020年4月郑州市第三人民医院收治的100例NSCLC患者为研究对象,免疫组化法检测并比较癌组织、癌旁组织中Keap1、Nrf2蛋白表达水平;比较不同临床病理参数患者Keap1、Nrf2蛋白表达水平;比较不同Keap1、Nrf2蛋白表达患者血清超氧化物歧化酶(SOD)、诱导型一氧化氮合酶(iNOS)、丙二醛(MDA)水平,并采用Spearman法分析SOD、i NOS、MDA与临床病理参数的相关性,采用Pearson法分析SOD、iNOS、MDA与Keap1、Nrf2蛋白水平的的相关性;比较不同Keap1、Nrf2蛋白表达患者的生存率。结果癌组织、癌旁组织Keap1蛋白阳性率分别为77.00%、53.00%,Nrf2蛋白阳性率分别为74.00%、45.00%,Keap1蛋白OD值分别为0.41±0.07、0.33±0.05,Nrf2蛋白OD值分别为0.39±0.06、0.31±0.06,癌组织Keap1、Nrf2蛋白阳性率及OD值明显高于癌旁组织,差异均有统计学意义(P<0.05);Keap1蛋白阳性表达与病理分级、T分期呈正相关(r=0.569、0.574,P<0.01),Nrf2蛋白阳性表达与病理分级、T分期呈正相关(r=0.527、0.539,P<0.01);Keap1蛋白阳性者、阴性者的血清SOD水平分别为(86.78±9.14)U/m L、(115.07±12.13)U/m L,MDA水平分别为(4.42±0.82)mmol/L、(3.24±0.56)mmol/L,i NOS水平分别为(22.74±4.31)U/m L、(15.59±3.02)U/mL,Nrf2蛋白阳性者、阴性者血清SOD水平分别为(84.94±9.12)U/mL、(117.06±12.37)U/mL,MDA水平分别为(4.48±0.85)mmol/L、(3.21±0.52)mmol/L,iNOS水平分别为(23.02±4.28)U/mL、(15.64±3.10)U/mL,Keap1、Nrf2蛋白阳性者血清SOD水平明显低于阴性者,MDA、iNOS水平明显高于阴性者,差异均有统计学意义(P<0.05);Keap1、Nrf2蛋白表达与SOD呈负相关(r=-0.612、-0.614,P<0.01),与MDA、iNOS呈正相关(r_(Keap1)=0.609、0.614,P<0.01;r_(Nrf2)=0.610、0.608,P<0.01);Keap1、Nrf2蛋白阳性表达者3年生存率为85.71%、83.78%,明显低于阴性表达者的95.65%、100.00%,差异均有统计学意义(P<0.05)。结论NSCLC组织中Keap1、Nrf2蛋白表达水平升高,且与病理分级、T分期密切相关,该信号通路活化可参与氧化应激反应过程,且对预判患者预后具有一定临床意义。

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。

Lactobacillus plantarum J26 alleviates alcohol-induced oxidative liver injury by regulating the Nrf2 signaling pathway

Oxidative stress is one of the main ways to cause alcohol-induced liver injury,and alcoholic liver disease(ALD)has been a common health problem worldwide.Lactic acid bacteria(LAB)is also considered as a potential treatment to alleviate alcohol-induced liver injury.Lactobacillus plantarum J26 is a LAB isolated from Chinese traditional fermented dairy products with excellent probiotic effects.This study aimed to establish a mice model of alcoholic liver injury through acute-on-chronic alcohol feeding and to study the alleviating effect of pre-intake of L.plantarum J26 on alcohol-induced oxidative liver injury and focus on its potential mechanism of alleviating effect.The results showed that pre-intake of L.plantarum J26 could improve liver pathological changes,reduce lipid accumulation,increase mitochondrial ATP and mitochondrial(mtDNA)levels,and alleviate liver injury.In addition,pre-intake L.plantarum J26 can improve the level of short-chain fatty acids(SCFAs)in the intestines in mice,short chain fatty acids can be used as a signaling molecule activation of nuclear factor E2-related factor 2(Nrf2)signaling pathway to alleviate liver oxidative stress,and maintain mitochondrial homeostasis by regulating the expression of genes related to mitochondrial dynamics and autophagy,thereby reducing cell apoptosis to alleviate alcohol-induced oxidative liver injury.

抑制核转录因子E2相关因子2途径对高糖状态下胰腺癌细胞转移及免疫逃逸因子表达的调节作用

目的:探究抑制核转录因子E2相关因子2(Nrf2)途径对高糖状态下胰腺癌细胞转移及分泌免疫逃逸因子水平的影响。方法:培养人胰腺癌细胞株Panc-1,采用5.5、10、25、50 mmol/L葡萄糖处理细胞,分别于12 h、24 h、48 h通过MTT检测细胞增殖率,24 h时通过RT-qPCR和Western blot检测细胞内Nrf2表达变化;实验分为:对照组、高糖(HG)组、Nrf2抑制剂ML385+高糖(ML385+HG)组,MTT检测细胞增殖率,细胞克隆形成实验检测细胞集落形成数,Transwell检测细胞迁移数与侵袭数,体外划痕实验检测细胞划痕愈合情况,ELISA测定细胞培养液上清中血管内皮生长因子(VEGF)、IFN-γ、转化生长因子-β1(TGF-β1)和IL-6含量,细胞免疫荧光染色观察细胞内Nrf2分布,Western blot测定细胞中Nrf2和血红素加氧酶-1(HO-1)蛋白表达。结果:相较于5.5 mmol/L葡萄糖组,10、25、50 mmol/L葡萄糖处理12 h和24 h时Panc-1细胞增殖率升高,Nrf2 mRNA和蛋白表达均升高(P<0.05);与对照组比较,HG组细胞增殖率升高,集落形成数增加,迁移数与侵袭数均增加,划痕愈合率升高,细胞培养上清中VEGF、IFN-γ、TGF-β1和IL-6含量均增加,Nrf2荧光染色明显增强,细胞核内Nrf2表达增加,Nrf2和HO-1蛋白表达上调(P<0.05);与HG组比较,ML385+HG组细胞增殖率降低,集落形成数、迁移数与侵袭数均减少,划痕愈合率下降,培养上清中VEGF、IFN-γ、TGF-β1和IL-6含量均减少,细胞内Nrf2荧光染色较弱,Nrf2和HO-1蛋白表达下调(P<0.05)。结论:高糖状态下胰腺癌细胞中Nrf2高表达,抑制Nrf2途径能够抑制高糖促进的胰腺癌细胞增殖、迁移及侵袭,并减少免疫逃逸因子分泌。

PCNA、Bcl-2及EGFR在喉癌组织中的表达及与临床病理特征、生存的关系

目的探讨增殖细胞核抗原(PCNA)、B淋巴细胞瘤-2(Bcl-2)及表皮生长因子受体(EGFR)在喉癌组织中的表达及与临床病理特征、生存的关系。方法选取2017年3月—2020年1月在苏州大学附属第一医院因喉癌行手术治疗的92例患者的喉癌组织及对应癌旁组织标本。检测癌组织与癌旁组织PCNA mRNA、Bcl-2mRNA、EGFR mRNA相对表达量,多元线性回归分析其癌组织表达与临床病理特征的关系。随访3年,采用Kaplain-Maier曲线分析不同PCNA、Bcl-2、EGFR表达水平患者生存情况差异。结果癌组织PCNA mRNA、Bcl-2 mRNA、EGFR mRNA相对表达量高于癌旁组织(P<0.05)。不同年龄、肿瘤部位患者PCNA mRNA、Bcl-2 mRNA、EGFR mRNA相对表达量比较,差异无统计学意义(P>0.05);低分化,临床分期Ⅲ、Ⅳ期及淋巴结转移患者PCNA mRNA、Bcl-2 mRNA、EGFR mRNA相对表达量分别高于中、高分化,临床分期Ⅰ、Ⅱ期,无淋巴结转移患者(P<0.05)。多元线性回归分析结果显示,肿瘤分化程度、临床分期、淋巴结转移是喉癌组织PCNA mRNA、Bcl-2 mRNA、EGFR mRNA表达的影响因素。Kaplain-Maier曲线分析结果显示,PCNA mRNA高表达患者3年无进展生存率、总生存率分别为59.57%和70.21%,低于低表达患者的80.00%和88.89%(P<0.05);Bcl-2 mRNA高表达患者3年无进展生存率、总生存率分别为60.78%和70.59%,低于低表达患者的80.49%和90.24%(P<0.05);EGFR mRNA高表达患者3年无进展生存率、总生存率分别为59.09%和70.45%,低于低表达患者的79.17%、87.50%(P<0.05)。结论喉癌组织PCNA、Bcl-2、EGFR呈高表达,且其高表达状态与肿瘤分期高、分化程度低、淋巴结转移有关,PCNA、Bcl-2、EGFR表达水平可在一定程度上反映患者预后。

胃衡汤调控Nrf2-Keap信号通路抑制老年胃癌前病变患者氧化应激反应的临床研究

目的探讨胃衡汤调控核因子E2相关因子2(Nrf2)-Kelch样环氧氯丙烷相关蛋白(Keap)信号通路抑制老年胃癌前病变(Precancerous lesions of gastric cancer,PLGC)患者氧化应激反应的作用。方法选取2022年3月-2023年3月期间于南京中医药大学第二附属医院消化腔镜中心和脾胃病科就诊的PLGC老年患者60例,按随机数字表法分为基础治疗组与胃衡汤治疗组,每组各30例。另选择同期常规胃镜体检的慢性浅表性胃炎患者30例为对照组。对照组仅入组接受研究相关指标检测,不予治疗。基础治疗组患者给予雷贝拉唑钠肠溶胶囊,胃衡汤组在基础治疗组上加用胃衡汤治疗,均治疗6个月。观察比较两组PLGC患者临床疗效、安全性,治疗前后中医证候评分、胃镜病理分级及Nrf2-Keap通路相关蛋白检测。结果治疗后胃衡汤治疗组中医证候疗效总有效率93.33%(28/30)明显高于基础治疗组76.67%(23/30),差异有统计学意义(P<0.05)。治疗后胃衡汤治疗组病理组织学疗效总有效率83.33%(25/30)明显高于基础治疗组63.33%(19/30),差异有统计学意义(P<0.05)。治疗后两组患者中医证候各项积分及总积分均较治疗前降低,差异有统计学意义(P<0.05);且胃衡汤治疗组中医证候各项积分及总积分均明显低于基础治疗组,差异有统计学意义(P<0.05)。治疗后两组患者胃黏膜组织学各项积分及总分均较治疗前降低,差异有统计学意义(P<0.05);且胃衡汤治疗组胃黏膜组织学各项积分及总分均明显低于基础治疗组,差异有统计学意义(P<0.05)。治疗后两组患者Nrf2和SOD表达高于治疗前,Keap1和MDA表达较治疗前降低,差异有统计学意义(P<0.05);且胃衡汤治疗组Nrf2和SOD表达明显高于基础治疗组,Keap1和MDA表达明显低于基础治疗组,差异有统计学意义(P<0.05)。治疗期间,两组患者治疗前后肝肾功能、三大常规及心电图检查均未发现异常。结论胃衡汤治疗老年PLGC能够改善临床症状,抑制甚至逆转病理学改变,提高治疗效果,其机制与调节Nrf2-Keap信号通路而抑制氧化应激反应有关。

Aberrant activation of nuclear factor of activated T cell 2 in lamina propria mononuclear cells in ulcerative colitis

AIM:To investigate the role of nuclear factor of activated T cell 2(NFAT2),the major NFAT protein in peripheral T cells,in sustained T cell activation and intractable inflammation in human ulcerative colitis(UC). METHODS:We used two-dimensional gel-electrophoresis, immunohistochemistry,double immunohistochemical staining,and confocal microscopy to inspect the expression of NFAT2 in 107,15,48 and 5 cases of UC, Crohn's disease(CD),non-specific colitis,and 5 healthy individuals,respectively. RESULTS:Up-regulation with profound nucleo- translocation/activation of NFAT2 of lamina propria mononuclear cells(LPMC)of colonic mucosa was found specifically in the affected colonic mucosa from patients with UC,as compared to CD or NC(P<0.001,Kruskal- Wallis test).Nucleo-translocation/activation of NFAT2 primarily occurred in CD8+T,but was less prominent in CD4+T cells or CD20+B cells.It was strongly associated with the disease activity,including endoscopic stage (τ=0.2145,P=0.0281)and histologic grade(τ=0.4167, P<0.001). CONCLUSION:We disclose for the first time the nucleo-translocation/activatin of NFAT2 in lamina propria mononuclear cells in ulcerative colitis.Activation of NFAT2 was specific for ulcerative colitis and highly associated with disease activity.Since activation of NFAT2is implicated in an auto-regulatory positive feedback loop of sustained T-cell activation and NFAT proteins play key roles in the calcium/calcineurin signaling pathways,our results not only provide new insights into the mechanism for sustained intractable inflammation,but also suggest the calcium-calcineurin/NFAT pathway as a new therapeutic target for ulcerative colitis.

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.

Role of nuclear factor (erythroid-derived 2)-like 2 in metabolic homeostasis and insulin action: A novel opportunity for diabetes treatment?

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 inflammation, aging and cancer. Thus, the capacity of endogenous free radical clearance can be of patho-physiological importance; in this regard, the major reactive oxygen species defense machinery, the nuclear factor (erythroid-derived 2)-like 2 (Nrf2) system needs to be precisely 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.

Sinapic acid ameliorates D-galactosamine/lipopolysaccharideinduced fulminant hepatitis in rats:Role of nuclear factor erythroidrelated factor 2/heme oxygenase-1 pathways

BACKGROUND Sinapic acid(SA)has been shown to have various pharmacological properties such as antioxidant,antifibrotic,anti-inflammatory,and anticancer activities.Its mechanism of action is dependent upon its ability to curb free radical production and protect against oxidative stress-induced tissue injuries.AIM To study the hepatoprotective effects of SA against lipopolysaccharide(LPS)/Dgalactosamine(D-GalN)-induced acute liver failure(ALF)in rats.METHODS Experimental ALF was induced with an intraperitoneal(i.p.)administration of 8μg LPS and 800 mg/kg D-GalN in normal saline.SA was administered orally once daily starting 7 d before LPS/D-GalN treatment.RESULTS Data showed that SA ameliorates acute liver dysfunction,decreases serum levels of alanine transaminase(ALT),and aspartate aminotransferase(AST),as well as malondialdehyde(MDA)and NO levels in ALF model rats.However,pretreatment with SA(20 mg/kg and 40 mg/kg)reduced nuclear factor kappalight-chain-enhancer of activated B cells(NF-κB)activation and levels of inflammatory cytokines(tumor necrosis factor-αand interleukin 6).Also,SA increased the activity of the nuclear factor erythroid-related factor 2/heme oxygenase-1(Nrf2/HO-1)signaling pathway.CONCLUSION In conclusion,SA offers significant protection against LPS/D-GalN-induced ALF in rats by upregulating Nrf2/HO-1 and downregulating NF-κB.

基于Nrf2/ARE信号通路探讨夹脊电针对神经根型颈椎病模型大鼠的作用机制

目的:观察夹脊电针对神经根型颈椎病(CSR)模型大鼠的作用效应,从Nrf2/ARE通路探究其作用机制。方法:将75只SPF级SD大鼠随机分为空白对照组、假手术组、模型组、针刺组和电针组。空白对照组不予任何处理,用手术方法将用福尔马林溶液浸泡过的定量滤纸片放在C 8神经根腋下建立CSR型,假手术组仅暴露神经根,不放置滤纸片。造模完成后观察各组大鼠的疼痛行为表现、步态障碍评分和抓握能力评分,从造模后第4天开始,假手术组和模型组予以每日捆绑15 min,针刺组予以每日捆绑+针刺15 min,电针组予以每日捆绑+电针15 min。治疗结束后用试剂盒检测大鼠血清中丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-PX)的含量或活力,运用Western blot技术检测大鼠神经根组织中核因子E2相关因子(Nrf2)、抗体Kelch样ECH相关蛋白(Keap1)和血红素氧合酶(HO-1)的表达。结果:针刺组和电针组大鼠疼痛行为表现、步态障碍和抓握能力改善程度和好转速度均优于模型组。与空白对照组比较,假手术组大鼠血清MDA的含量、SOD与GSH-PX的活性差异无统计学意义(P>0.05)。与假手术组比较,模型组大鼠血清MDA的含量显著升高,SOD与GSH-PX的活性显著降低,并且神经根组织Nrf2与HO-1表达显著下降,Keap1表达显著上升,差异具有统计学意义(P<0.01)。与模型组比较,针刺组和电针组大鼠血清MDA的含量明显降低,SOD的活性明显升高,差异具有统计学意义(P<0.05),GSH-PX的活性显著升高,差异具有统计学意义(P<0.01),神经根组织Nrf2与HO-1的表达显著上升,Keap1的表达显著下降,差异具有统计学意义(P<0.01)。与针刺组比较,电针组大鼠血清MDA的含量明显升高,SOD与GSH-PX的活性明显降低,差异具有统计学意义(P<0.05),神经根组织Nrf2的表达显著上升,差异具有统计学意义(P<0.01),HO-1的表达明显上升,差异具有统计学意义(P<0.05),Keap1的表达显著下降,差异具有统计学意义(P<0.01)。结论:电针夹脊穴可以改善CSR大鼠的步态障碍,减轻CSR大鼠的根性疼痛,其机制可能与夹脊电针激活Nrf2/ARE信号通路,上调Nrf2、HO-1的表达和下调Keap1的表达水平有关。

灵芝提取物通过Nrf2/ARE通路对肝硬化小鼠肝功能的保护作用研究

[目的]探讨灵芝提取物(ganoderma lucidum extract,GLE)对肝硬化小鼠的肝保护作用及机制。[方法]将10只雄性C57BL/6小鼠作为对照组,剩余40只小鼠采用四氯化碳橄榄油混悬液诱导肝硬化模型,并随机分为模型组和GLE低(50 mg/kg·d)、中(100 mg/kg·d)、高(200 mg/kg·d)剂量组,对照组及模型组均灌胃等量0.9%氯化钠溶液。计算肝脏指数;以全自动生化分析仪检测小鼠血清中谷氨酸转氨酶(alanine aminotransferase,ALT)、天冬氨酸转氨酶(aspartate transaminase,AST)活性和总胆固醇(total cholesterol,TC)、总胆红素(total bilirubin,TB)和肌酐(creatinine,Cr)水平;以苏木精-伊红(hematoxylin eosin,HE)染色观察肝组织病理学变化,Masson染色观察肝组织纤维化程度;以脱氧核苷酸末端转移酶介导的dUTP缺口末端标记(terminal-deoxynucleotidyl transferase mediated dUTP nick end labeling,TUNEL)染色观察肝细胞凋亡情况;酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、IL-6、丙二醛(malondialdehyde,MDA)水平和超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性;免疫印迹法检测肝组织中总核因子E2相关因子2(nuclear factor E2 related factor 2,Nrf2)和核Nrf2、血红素加氧酶-1(heme oxygenase-1,HO-1)及醌氧化还原酶1[NAD(P)H:quinone oxidoreductase 1,NQO1]、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、Ⅰ型胶原蛋白(CollagenⅠ)及E-钙黏蛋白(E-cadherin)的表达情况。[结果]与对照组比较,模型组小鼠肝损伤明显,肝脏指数,血清ALT、AST活性,TC、TB及Cr水平,肝纤维化程度,肝细胞凋亡指数,血清TNF-α、IL-1β、IL-6及MDA水平,α-SMA及CollagenⅠ蛋白相对表达量升高(P<0.05),血清SOD和GSH-Px活性、肝组织总Nrf2和核Nrf2、HO-1、NQO1及E-cadherin蛋白表达降低(P<0.05)。与模型组比较,GLE低、中、高剂量组小鼠肝损伤逐步减轻,肝脏指数,血清ALT、AST活性,TC、TB及Cr水平,肝纤维化程度,肝细胞凋亡指数,血清TNF-α、IL-1β、IL-6及MDA水平,α-SMA及CollagenⅠ蛋白表达降低(P<0.05),血清SOD和GSH-Px活性、肝组织总Nrf2和核Nrf2、HO-1、NQO1及E-cadherin蛋白表达升高(P<0.05)。[结论]GLE可减轻肝硬化小鼠组织病理损伤,改善肝功能,这可能与激活Nrf2/ARE通路,抑制氧化应激和炎症反应,进而干预肝纤维化有关。