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.

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 of miR-27b-3p and Nrf2 in human retinal pigment epithelial cell induced by high-glucose

AIM:To determine whether the microRNA-27b-3p(miR-27b-3p)/NF-E2-related factor 2(Nrf2)pathway plays a role in human retinal pigment epithelial(hRPE)cell response to high glucose,how miR-27b-3p and Nrf2 expression are regulated,and whether this pathway could be specifically targeted.METHODS:hRPE cells were cultured in normal glucose or high glucose for 1,3,or 6d before measuring cellular proliferation rates using cell counting kit-8 and reactive oxygen species(ROS)levels using a dihydroethidium kit.miR-27b-3p,Nrf2,NAD(P)H quinone oxidoreductase 1(NQO1)and heme oxygenase-1(HO-1)mRNA and protein levels were analyzed using reverse transcription quantitative polymerase chain reaction(RT-qPCR)and immunocytofluorescence(ICF),respectively.Western blot analyses were performed to determine nuclear and total Nrf2 protein levels.Nrf2,NQO1,and HO-1 expression levels by RT-qPCR,ICF,or Western blot were further tested after miR-27b-3p overexpression or inhibitor lentiviral transfection.Finally,the expression level of those target genes was analyzed after treating hRPE cells with pyridoxamine.RESULTS:Persistent exposure to high glucose gradually suppressed hRPE Nrf2,NQO1,and HO-1 mRNA and protein levels and increased miR-27b-3p mRNA levels.High glucose also promoted ROS release and inhibited cellular proliferation.Nrf2,NQO1,and HO-1 mRNA levels decreased after miR-27b-3p overexpression and,conversely,both mRNA and protein levels increased after expressing a miR-27b-3p inhibitor.After treating hRPE cells exposed to high glucose with pyridoxamine,ROS levels tended to decreased,proliferation rate increased,Nrf2,NQO1,and HO-1 mRNA and protein levels were upregulated,and miR-27b-3p mRNA levels were suppressed.CONCLUSION:Nrf2 is a downstream target of miR-27b-3p.Furthermore,the miR-27b-3p inhibitor pyridoxamine can alleviate high glucose injury by regulating the miR-27b-3p/Nrf2 axis.

核因子红系2相关因子3高表达与胶质瘤预后及迁移能力的关系

目的分析核因子红系2相关因子3(NFE2L3)在胶质瘤中的表达水平,及其与患者预后、胶质瘤迁移能力的关系。方法利用肿瘤基因组图谱(TCGA)数据库、中国胶质瘤基因组图谱(CGGA)数据库、基因表达谱交互分析(GEPIA)数据库和人类蛋白质图谱(HPA)数据库确定NFE2L3在各级别胶质瘤中的表达水平及其与患者预后的关系。构建NFE2L3-siRNA载体,并在胶质瘤U251、T98细胞系中进行转染。采用实时荧光定量PCR和Western blot检测转染NFE2L3-siRNA后胶质瘤U251和T98细胞中NFE2L3和MMP-9的表达水平。使用细胞划痕实验和transwell迁移实验检测NFE2L3对U251、T98细胞迁移功能的影响。结果NFE2L3在人脑胶质瘤组织中高表达,其表达水平越高,患者预后越差(P<0.001)。NFE2L3的表达水平与患者年龄、组织学类型、化疗状态、1p19q编码缺失状态、IDH相关突变状态密切相关(P<0.001)。在U251细胞和T98细胞中,干扰组的迁移能力、MMP-9在mRNA和蛋白水平的表达均低于对照组(P<0.01)。结论NFE2L3的高表达与胶质瘤患者的预后不良有关,NFE2L3的敲低显著抑制U251、T98细胞的迁移能力。NFE2L3促进胶质瘤U251、T98细胞的迁移,NFE2L3有可能是临床治疗胶质瘤的潜在靶标。

芍药苷预处理对大鼠心肌缺血-再灌注损伤氧化应激的保护作用及对Nrf-2/TXNIP/NLRP-3信号通路的影响

目的探讨芍药苷(PF)对心肌缺血-再灌注损伤(MIRI)大鼠氧化应激的影响及其可能作用机制。方法选取65只SD大鼠采用结扎冠状动脉(冠脉)左前降支建立MIRI模型,造模成功后随机分为模型组、PF低、高剂量(60 mg/kg、120 mg/kg)组和西药(1 mg/kg盐酸地尔硫卓)组各15只,另设对照组。检测大鼠天冬氨酸氨基转移酶(AST)、肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)水平;HE染色观察心肌组织病理学变化;RT-PCR和Western blot检测心肌组织核因子E2相关因子2(Nrf2)、硫氧还蛋白互作蛋白(TXNIP)、NOD样受体蛋白3(NLRP-3)mRNA和蛋白表达情况。结果与模型组比较,各实验组AST、CK、CK-MB、LDH、MDA水平、TXNIP、NLRP-3 mRNA和蛋白水平降低,GSH-Px、SOD、CAT水平、Nrf2 mRNA和蛋白水平升高(P<0.05)。与PF低剂量组比较,PF高剂量组以上指标效果更佳(P<0.05)。结论PF可减轻MIRI大鼠氧化应激,改善心肌损伤,可能与Nrf2/TXNIP/NLRP-3信号通路有关。

结直肠癌组织的NFE2L3P1水平及临床意义

目的探讨结直肠癌组织中核因子E2相关因子3(NFE2L3)及其假基因1(NFE2L3P1)的水平及临床意义。方法采用实时定量PCR检测89对结直肠癌组织和癌旁组织的NFE2L3和NFE2L3P1水平并分析两者的相关性,评价NFE2L3P1水平与结直肠癌临床病理特征的关系,根据随访数据结合Kaplan-Meier生存曲线法分析不同NFE2L3P1水平对患者预后的影响。结果结直肠癌组织的NFE2L3水平高于癌旁组织(1.899±0.835 vs.1.275±0.526),NFE2L3P1水平亦高于癌旁组织(3.433±1.192 vs.1.261±0.742),差异有统计学意义(P<0.05),且NFE2L3水平和NFE2L3P1水平呈正相关(r=0.608,P<0.001)。分层分析显示组织NFE2L3P1水平与分化程度(χ^(2)=6.273,P=0.012)、淋巴结转移(χ^(2)=4.094,P=0.043)、T分期(χ^(2)=4.022,P=0.045)和TNM分期(χ^(2)=6.059,P=0.014)有关。单因素分析发现分化程度(χ^(2)=6.424,P=0.011)、肿瘤大小(χ^(2)=5.051,P=0.025)、TNM分期(χ^(2)=4.052,P=0.044)和NFE2L3P1水平(χ^(2)=6.020,P=0.014)与结直肠癌的总生存期(OS)有关,其中NFE2L3P1低水平者的中位OS为58.0(95%CI:54.551~61.449)个月,优于高水平者的52.0(95%CI:47.152~56.848)个月(P<0.05)。结论NFE2L3P1在结直肠癌中高表达,在结直肠癌的进展和预后中起着重要作用,可能是预测切除术后生存率的有用生物标志物。

High glucose reduces Nrf2-dependent cRAGE release and enhances inflammasome-dependent IL-1βproduction in monocytes:the modulatory effects of EGCG

Soluble receptor for advanced glycation end products(sRAGE)acts as a decoy sequestering of RAGE ligands,thus preventing the activation of the ligand-RAGE axis linking human diseases.However,the molecular mechanisms underlying sRAGE remain unclear.In this study,THP-1 monocytes were cultured in normal glucose(NG,5.5 mmol/L)and high glucose(HG,15 mmol/L)to investigate the effects of diabetesrelevant glucose concentrations on sRAGE and interleukin-1β(IL-1β)secretion.The modulatory effects of epigallocatechin gallate(EGCG)in response to HG challenge were also evaluated.HG enhanced intracellular reactive oxygen species(ROS)generation and RAGE expression.The secretion of sRAGE,including esRAGE and cRAGE,was reduced under HG conditions,together with the downregulation of a disintegrin and metallopeptidase 10(ADAM10)and nuclear factor erythroid 2-related factor 2(Nrf2)nuclear translocation.Mechanistically,the HG effects were counteracted by siRAGE and exacerbated by siNrf2.Chromatin immunoprecipitation results showed that Nrf2 binding to the ADAM10 promoter and HG interfered with this binding.Our data reinforce the notion that RAGE and Nrf2 might be sRAGE-regulating factors.Under HG conditions,the treatment of EGCG reduced ROS generation and RAGE activation.EGCG-stimulated cRAGE release was likely caused by the upregulation of the Nrf2-ADAM10 pathway.EGCG inhibited HG-mediated NLRP3 inflammasome activation at least partly by stimulating sRAGE,thereby reducing IL-1βrelease.

Hydrogen sulfide reduces oxidative stress in Huntington's disease via Nrf2

The pathophysiology of Huntington's disease involves high levels of the neurotoxin quinolinic acid. Quinolinic acid accumulation results in oxidative stress, which leads to neurotoxicity. However, the molecular and cellular mechanisms by which quinolinic acid contributes to Huntington's disease pathology remain unknown. In this study, we established in vitro and in vivo models of Huntington's disease by administering quinolinic acid to the PC12 neuronal cell line and the striatum of mice, respectively. We observed a decrease in the levels of hydrogen sulfide in both PC12 cells and mouse serum, which was accompanied by down-regulation of cystathionine β-synthase, an enzyme responsible for hydrogen sulfide production. However, treatment with NaHS(a hydrogen sulfide donor) increased hydrogen sulfide levels in the neurons and in mouse serum, as well as cystathionine β-synthase expression in the neurons and the mouse striatum, while also improving oxidative imbalance and mitochondrial dysfunction in PC12 cells and the mouse striatum. These beneficial effects correlated with upregulation of nuclear factor erythroid 2-related factor 2 expression. Finally, treatment with the nuclear factor erythroid 2-related factor 2inhibitor ML385 reversed the beneficial impact of exogenous hydrogen sulfide on quinolinic acid-induced oxidative stress. Taken together, our findings show that hydrogen sulfide reduces oxidative stress in Huntington's disease by activating nuclear factor erythroid 2-related factor 2,suggesting that hydrogen sulfide is a novel neuroprotective drug candidate for treating patients with Huntington's disease.

Procyanidin A_1 and its digestive products alleviate acrylamide-induced IPEC-J2 cell damage through regulating Keap1/Nrf2 pathway

Our previous study has revealed that procyanidin A_(1)(A_(1))and its simulated digestive product(D-A,)can alleviate acrylamide(ACR)-induced intestine cell damage.However,the underlying mechanism remains unknown.In this study,we elucidated the molecular mechanism for and D-A_(1) to alleviate ACR-stimulated IPEC-J2 cell damage.ACR slightly activated nuclear factor erythroid 2-related factor 2(Nrf2)signaling and its target genes,but this activation could not reduce intestine cell damage.A_(1) and D-A_(1) could alleviate ACR-induced cell damage,but the effect was abrogated in cells transiently transfected with Nrf2 small interfering RNA(siRNA).Further investigation confirmed that A_(1) and D-A_(1) interacted with Ketch-like ECH-associated protein 1(Keapl),which boosted the stabilization of Nrf2,subsequently promoted the translocation of Nrf2 into the nucleus,and further increased the expression of antioxidant proteins,thereby inhibiting glutathione(GSH)consumption,maintaining redox balance and eventually alleviating ACR-induced cell damage.Importantly,there was no difference between A_(1) and D-A_(1) treated groups,indicating that A_(1) can tolerate gastrointestinal digestion and may be a potential compound to limit the toxicity of ACR.

Functionalized selenium nanoparticles ameliorated acetaminophen-induced hepatotoxicity through synergistically triggering PKCδ/Nrf2 signaling pathway and inhibiting CYP 2E1

Selenium nanoparticles(SeNPs)have been demonstrated potential for use in diseases associated with oxidative stress.Functionalized SeNPs with lower toxicity and higher biocompatibility could bring better therapeutic activity and clinical application value.Herein,this work was conducted to investigate the protective effect of Pleurotus tuber-regium polysaccharide-protein complex funtionnalized SeNPs(PTR-SeNPs)against acetaminophen(APAP)-induced oxidative injure in HepG2 cells and C57BL/6J mouse liver.Further elucidation of the underlying molecular mechanism,in particular their modulation of Nrf2 signaling pathway was also performed.The results showed that PTR-SeNPs could significantly ameliorate APAP-induced oxidative injury as evidenced by a range of biochemical analysis,histopathological examination and immunoblotting study.PTR-SeNPs could hosphorylate and activate PKCδ,depress Keap1,and increase nuclear accumulation of Nrf2,resulting in upregulation of GCLC,GCLM,HO-1 and NQO-1 expression.Besides,PTR-SeNPs suppressed the biotransformation of APAP to generate intracellular ROS through CYP 2E1 inhibition,restoring the mitochondrial morphology.Furthermore,the protective effect of PTR-SeNPs against APAP induced hepatotoxicity was weakened as Nrf2 was depleted in vivo,indicating the pivotal role of Nrf2 signaling pathway in PTR-SeNPs mediated hepatoprotective efficacy.Being a potential hepatic protectant,PTR-SeNPs could serve as a new source of selenium supplement for health-promoting and biomedical applications.

Curcumin Inhibits Lipopolysaccharide-lnduced Mucin 5AC Hypersecretion and Airway Inflammation via Nuclear Factor Erythroid 2-Related Factor 2

Background： Excess mucus production is an important pathophysiological feature of chronic inflammatory airway diseases. Effective therapies are currently lacking. The aim of the study was to evaluate the effects ofcurcumin （CUR） on lipopolysaccharide （LPS）-induced mucus secretion and inflammation, and explored the underlying mechanism in vivo and in vitro. Methods： For the in vitro study, human bronchial epithelial （NCI-H292） cells were pretreated with CUR or vehicle for 30 min, and then exposed to LPS for 24 h. Next, nuclear factor erythroid 2-related factor 2 （Nrf2） was knocked down with Nrf2 small interfering RNA （siRNA） to confirm the specific role of Nrf2 in mucin regulation of CUR in NCI-H292 cells. In vivo, C57BL/6 mice were randomly assigned to three groups （n = 7 for each group）： control group, LPS group, and LPS ＋ CUR group. Mice in LPS and LPS ＋ CUR group were injected with saline or CUR （50 mg/kg） intraperitoneally 2 h before intratracheal instillation with LPS （ 100 μg/ml） for 7 days. Cell lysate and lung tissue were obtained to measured Mucin 5AC （MUC5AC） and Nrf2 mRNA and protein expression by a real-time polymerase chain reaction and Western blotting. Bronchoalveolar lavage fluid （BALF） was collected to enumerate total cells and neutrophils. HistopathologicaI changes of the lung were observed. Data were analyzed by one-way analysis of variance. Student＇s t-test was used when two groups were compared. Results： CUR significantly decreased the expression ofMUC5AC mRNA and protein in NCI-H292 cells exposed to LPS. This effect was dose dependent （2.424 ± 0.318 vs. 7.169 ± 1.785, t = 4.534, and 1.060 ± 0.197 vs. 2.340 ± 0.209, t = 7.716; both P 〈 0.05, respectively） and accompanied by increased mRNA and protein expression of Nrf2 （1.952 ± 0.340 vs. 1.142 ± 0.176, t = -3.661, and 2.010 ± 0.209 vs. 1.089 ±0. 132, t = -6.453; both P 〈 0.05, respectively）. Furthermore, knockdown of Nrf2 with siRNA increased MUC5A C mRN A expression by 47.7%, compared with levels observed in the siRNA-negative group （6.845 ± 1.478 vs. 3.391 ± 0.517, t = -3.821, P 〈 0.05）. Knockdown of Nrf2 with siRNA also markedly increased MUC5A C protein expression in NCI-H292 cells. CUR also significantly decreased LPS-induced mRNA and protein expression of MUC5A C in mouse lung （ 1.672 ± 0.721 vs. 5.961 ± 2.452, t = 2.906, and 0.480 ± 0.191 vs. 2.290 ± 0.834, t = 3.665, respectively; both P 〈 0.05）. Alcian blue/periodic acid-Schiff staining also showed that CUR suppressed mucin production. Compared with the LPS group, the numbers of inflammatory cells （247 ± 30 vs. 334 ± 24, t = 3.901, P 〈 0.05） and neutrophils （185 ± 22 vs. 246 ± 20, t = 3.566, P 〈 0.05） in BALF decreased in the LPS ＋ CUR group, as well as reduced inflammatory cell infiltration in lung tissue. Conclusion： CUR inhibits LPS-induced airway mucus hypersecretion and inflammation through activation of Nrf2 possibly.

虾青素保护PC-3细胞免受H_2O_2氧化应激的作用机制

目的:研究虾青素对过氧化氢诱导PC-3细胞氧化应激的保护作用,探索其信号通路机制。方法:建立H2O2氧化应激模型,采用不同浓度虾青素预处理PC-3细胞,检测细胞存活率、细胞凋亡、活性氧(reactive oxygen species,ROS)水平、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2-associated X protein,Bax)、活化半胱天冬酶-3表达及丝裂原活化蛋白激酶-核因子E2相关基因2-血红素氧合酶1(mitogen-activated protein kinases nuclear factor erythroid-2-related factor 2-heme oxygenase 1,MAPK-Nrf2-HO-1)通路的变化。结果:20μmol/L虾青素预处理显著提高H2O2所降低的细胞存活率、降低ROS水平(P<0.05),同时通过抑制Bcl-2/Bax比率下降及半胱天冬酶-3的激活,从而使细胞凋亡率从51.4%降低至14.8%,进一步研究发现虾青素能够促进Nrf2磷酸化,并促进HO-1的表达,呈现浓度依赖性。通过细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)抑制剂(U0126)和Akt抑制剂(LY294002)预处理,发现当ERK和磷脂酰肌醇激酶/蛋白激酶B(phosphoinositide 3-kinase/protein kinase B,PI3K/Akt)通路被抑制后,Nrf2表达降低,表明HO-1上调受上游ERK和胞内PI3K/Akt通路的调控。在对MAPK途径对细胞毒性影响的研究中,ERK通路被抑制后细胞存活率显著下降,而c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)和p38 MAPK通路被抑制后并不影响其保护作用,表明虾青素抑制细胞存活率下降是通过MAPK途径中的ERK通路,而不是JNK和p38通路。结论:虾青素预处理PC-3细胞可以减轻H2O2诱导的氧化应激,维持细胞生理活性。

螺内酯对缺氧/复氧处理的人心肌细胞HO-1/Nrf2/SIRT3信号通路及自噬的影响

目的探讨螺内酯对缺氧/复氧(H/R)处理的人心肌细胞血红素加氧酶1(HO-1)/核因子E2相关因子2(Nrf2)/沉默信息调节因子3(SIRT3)信号通路及自噬的影响。方法体外培养人心肌细胞AC16,设置对照组(AC16细胞正常培养)、H/R组(AC16细胞经H/R处理)、低剂量螺内酯组(AC16细胞经H/R处理+0.5μmol/L螺内酯)、中剂量螺内酯组(AC16细胞经H/R处理+1μmol/L螺内酯)和高剂量螺内酯组(AC16细胞经H/R处理+1.5μmol/L螺内酯)。流式细胞仪检测各组AC16细胞凋亡情况;相应试剂盒检测各组AC16细胞中超氧化物歧化酶(SOD)、丙二醛(MDA)水平;实时荧光定量PCR(qRT-PCR)法检测各组AC16细胞中HO-1、Nrf2、SIRT3 mRNA表达情况;蛋白印迹(Western blot)法检测各组AC16细胞中HO-1、Nrf2、SIRT3、微管相关蛋白1轻链3(LC3)Ⅰ、LC3Ⅱ蛋白表达情况。结果与对照组比较,H/R组AC16细胞凋亡率、MDA含量、LC3Ⅰ蛋白表达水平及LC3Ⅰ/LC3Ⅱ比值明显升高(P<0.05),SOD活性,HO-1、Nrf2、SIRT3 mRNA及蛋白表达水平和LC3Ⅱ蛋白表达水平明显降低(P<0.05);随螺内酯剂量的升高,AC16细胞凋亡率、MDA含量、LC3Ⅰ蛋白表达水平及LC3Ⅰ/LC3Ⅱ比值明显降低(P<0.05),SOD活性,HO-1、Nrf2、SIRT3 mRNA及蛋白表达水平和LC3Ⅱ蛋白表达水平明显升高(P<0.05)。结论螺内酯可能通过激活HO-1/Nrf2/SIRT3信号通路及自噬,减少经H/R处理后的人心肌AC16细胞凋亡,发挥心肌保护作用。

CTHRC1和NFE2L3在结直肠癌组织中的表达及相关性

目的探讨CTHRC1和NFE2L3在结直肠癌早期诊断的意义。方法收集本院2005～2011年经过病理组织学检查诊断结直肠癌的手术标本51例与非癌症的经过病理组织学检查为正常结肠和直肠组织20例,应用免疫组化检测人结直肠中CTHRC1和NFE2L3蛋白的表达情况。结果癌组织中CTHRC1表达阳性47例(92.16%),正常组织中阳性1例(5.00%),两组比较差异有统计学意义(χ2=30.13,P<0.01);NFE2L3的瘤组织阳性43例(84.43%),正常组织中6例(30.00%),两组比较差异有统计学意义(χ2=23.22,P<0.01)。CTHRC1阳性表达组中,NFE2L3的阳性表达率为72.55%(37/51);在CTHRC1阴性表达组中,NFE2L3阳性表达率为1.96%(1/51),差异有统计学意义(χ2=36.54,r:0.510;P<0.01),两者表达呈正相关。结论 CTHRC1是结直肠癌发展过程中的重要因素,而NFE2L3可以直接调节CTHRC1基因的表达,这说明通过抑制NFE2L3的活性可以减少CTHRC1的表达,进而延缓结直肠癌的进展,从而发挥其抗肿瘤的作用。

紫草素通过调控PI3K/AKT通路减轻脑缺血再灌注损伤的机制研究

目的探讨脑缺血再灌注损伤后PI3K/AKT信号通路的活性变化及紫草素发挥相应作用的调控机制。方法采用改良的Longa线栓法制作小鼠大脑中动脉缺血再灌注模型。成年雄性CD-1小鼠随机被分为5组:假手术组(Sham)、缺血再灌注组(Vehicle)、低剂量紫草素干预组(L-Shi)、高剂量紫草素干预组(H-Shi)、紫草素+LY294002组(H-Shi+LY294002)。各组小鼠取材前先进行神经功能评分、取材后进行脑梗死体积、脑组织含水量的测定,并对超氧化物歧化酶(SOD)的活性和丙二醛(MDA)含量的变化进行检测,运用免疫组化,蛋白印迹法和实时定量PCR等方法检测缺血侧脑组织中PI3K、AKT、HO-1、Nrf2蛋白和mRNA水平的变化;以及紫草素在脑缺血再灌注损伤中的神经保护作用及其对PI3K/AKT信号转导通路的作用。结果H-Shi组可明显改善神经功能缺损、降低脑含水量、减小脑梗死体积;免疫组化、Western blot结果显示H-Shi组在24 h和72 h均可明显升高p-PI3K、p-AKT、HO-1、Nrf2的表达,但这种作用同样可被LY294002阻断。H-Shi对于HO-1、Nrf2的mRNA表达与免疫组化和Western blot结果一致。结论PI3K/AKT通路参与了紫草素的脑保护作用,为其临床进一步应用提供了理论基础。

高糖环境下PLGF基因沉默对人绒毛膜外滋养细胞增殖、迁移及侵袭及Nrf2/HO-1信号通路的影响

目的探究高糖环境下胎盘生长因子(PLGF)基因沉默对人绒毛膜外滋养细胞(EVCT)的影响。方法体外培养HTR-8/SVneo细胞,将细胞分为对照组(不做处理)、空载组(空载质粒转染)及PLGF沉默组(PLGF双切口酶质粒转染)。CCK-8法、划痕实验及Transwell实验检测各组细胞增殖、迁移、侵袭能力;q RT-PCR及Western blot检测各组细胞PLGF、核转录因子红系2相关因子2(Nrf2)、血红素氧合酶1(HO-1)mRNA及蛋白表达情况。结果对照组与空载组细胞增殖抑制率、划痕愈合率、侵袭细胞数比较,差异无统计学意义(P>0.05);与对照组比较,PLGF沉默组细胞增殖抑制率升高,划痕愈合率、侵袭细胞数降低(P<0.05)。对照组与空载组PLGF、Nrf2及HO-1 m RNA及蛋白表达量比较,差异无统计学意义(P>0.05);与对照组比较,PLGF沉默组PLGF、Nrf2及HO-1m RNA及蛋白表达量降低(P<0.05)。结论高糖环境下PLGF基因沉默能够抑制EVCT增殖、迁移及侵袭,其作用机制可能与抑制Nrf2/HO-1信号通路活性有关。

GLS1通过激活Nrf2/HO-1轴抑制过氧化氢诱导的ARPE-19细胞氧化应激、自噬与凋亡

目的探究GLS1对过氧化氢诱导的ARPE-19细胞氧化应激、自噬与凋亡的影响及机制。方法ARPE-19细胞分为对照组、H_(2)O_(2)组、H_(2)O_(2)+Vector组、H_(2)O_(2)+GLS1组,对照组细胞不做任何处理,H_(2)O_(2)组细胞用200μmol/L H_(2)O_(2)诱导48h,H_(2)O_(2)+Vector组和H_(2)O_(2)+GLS1组细胞转染Vector和GLS1质粒后用200μmol/L H_(2)O_(2)诱导48 h,比色法测定各组细胞SOD、GSH和MDA浓度,透射电镜观察各组细胞自噬小体,流式细胞术检测各组细胞凋亡水平,Western blot检测各组细胞Nrf2、HO-1、LC3-Ⅱ、p62的表达。结果与对照组比较,H_(2)O_(2)组ARPE-19细胞SOD、GSH表达显著下降,MDA显著增加,自噬小体数目显著增加,细胞凋亡显著增加,LC3-Ⅱ表达显著上调,P62、抗核因子红系2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)、血红素氧合酶1(heme Oxygenase-1,HO-1)表达显著下调。与H_(2)O_(2)+Vector组比较,H_(2)O_(2)+GLS1组细胞SOD、GSH表达显著增加,MDA显著降低,自噬小体数目显著减少,细胞凋亡显著减少,LC3-Ⅱ表达显著下调,P62、Nrf2、HO-1表达显著上调。结论GLS1可抑制H_(2)O_(2)诱导的ARPE-19细胞氧化应激、自噬与凋亡,其机制为激活Nrf2/HO-1信号通路。