Targeting nuclear factor erythroid 2-related factor 2-regulated ferroptosis to treat nervous system diseases

By critically examining the work,we conducted a comprehensive bibliometric analysis on the role of nuclear factor erythroid 2-related factor 2(NRF2)in nervous system diseases.We also proposed suggestions for future bibliometric studies,including the integration of multiple websites,analytical tools,and analytical approaches,The findings presented provide compelling evidence that ferroptosis is closely associated with the therapeutic challenges of nervous system diseases.Targeted modulation of NRF2 to regulate ferroptosis holds substantial potential for effectively treating these diseases.Future NRF2-related research should not only focus on discovering new drugs but also on designing rational drug delivery systems.In particular,nanocarriers offer substantial potential for facilitating the clinical translation of NRF2 research and addressing existing issues related to NRF2-related drugs.

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.

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.

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.

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.

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.

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.

Increased serum levels of soluble CD146 and vascular endothelial growth factor receptor 2 in patients with exudative age-related macular degeneration

AIM: To investigate serum levels of soluble CD146(s CD146) and vascular endothelial growth factor receptor 2(VEGFR2) in patients with age-related macular degeneration(AMD). METHODS: Eighty-eight patients with exudative AMD and 45 sex-and age-matched healthy controls were enrolled in this study conducted in China. Serum samples was obtained from the patients with exudative AMD and from the controls. Serum sCD146 and VEGFR2 protein levels were measured using an enzyme-linked immunosorbent assay.RESULTS: We found that serum sCD146 and VEGFR2 protein levels were significantly higher in the patients with exudative AMD group than in the controls(t=3.859, P<0.001 and t=3.829, P<0.001, respectively). Serum sCD146 levels were significantly higher in patients with classic choroidal neovascularization(CNV) than in those with occult CNV(t=9.899, P<0.001). There was a significant difference in the trend for exudative AMD in the highest versus lowest quartile of circulating sCD146 levels(χ2=10.29, P=0.001). The receiver operating characteristic curve analysis showed that the area under the curve was 0.696 for s CD146(95%CI: 0.601-0.791) with an optimum diagnostic cut-off value of 157.16 ng/mL, a sensitivity of 55.7%, and a specificity of 82.2%.CONCLUSION: The serum sCD146 level increases and may be a biomarker for exudative AMD.

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.

Serum vascular endothelial growth factor receptor-2 and adropin levels in age-related macular degeneration

AIM： To investigate the serum levels of vascular endothelial growth factor receptor-2（VEGFR-2） and adropin in age-related macular degeneration（AMD）patients.·METHODS： Ninety-eight AMD patients were included in the study. Seventy-eight age- and sex-matched healthy volunteers were recruited as the control group.Fundus florescein angiography and optical coherence tomography were performed to assess the posterior segment details. Serum VEGFR-2 and adropin levels were measured using enzyme-linked immunosorbent assays and compared between the study groups.· RESULTS： AMD group had significantly increased foveal retinal thickness, serum LDL and HDL levels and significantly decreased subfoveal choroidal thickness（P =0.01, 0.047, 0.025 and 〈0.001, respectively）. Serum VEGFR-2level revealed a significant decrease in AMD patients compared to controls（26.48 ±6.44 vs 30.42 ±7.92 ng/m L,P 〈0.001）. There was an insignificant increase in serum adropin level in AMD patients（6.17±3.19 vs 5.79±2.71 ng/m L,P =0.4）. Serum level of VEGFR-2 in AMD patients had a significant negative correlation with foveal retinal thickness（r =-0.226, P =0.025） and a significant positive correlation with subfoveal choroidal thickness（r=0.2, P=0.048）.·CONCLUSION： The current study demonstrated that the decreased serum VEGFR-2 level may be considered in the development of AMD. Adropin does not seem to play a role in the pathogenesis of AMD.

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.

Etomidate protects retinal ganglion cells from hydrogen peroxide-induced injury via Nrf2/HO-1 pathway

AIM:To determine whether etomidate(ET)has a protective effect on retinal ganglion cells(RGCs)injured with hydrogen peroxide(H_(2)O_(2))and to explore the potential mechanism underlying the antioxidative stress effect of ET.METHODS:Cultured RGCs were identified by double immunofluorescent labeling of microtubule-associated protein 2 and Thy1.1.An injury model of H_(2)O_(2)-induced RGCs oxidative stress was established in vitro.Cells were pretreated with different concentrations of ET(1,5,and 10μmol/L)for 4h,followed by further exposure to H_(2)O_(2)at 1000μmol/L.Cell counting kit 8 and Annexin V/propidium iodide assays were applied to detect the viabilities and apoptosis rates of the RGCs at 12,24,and 48h after H_(2)O_(2)stimulation.The levels of nitric oxide,malondialdehyde,and glutathione in culture media were measured at these time points.Quantitative reverse transcription polymerase chain reaction(qRT-PCR)and Western blot were performed to observe the effects of ET on the messenger RNA and protein expression of inducible nitric oxide synthase(iNOS),nuclear factor erythroid 2-related factor 2(Nrf2),heme oxygenase 1(HO-1),glutathione peroxidase 1 and the level of conjugated acrolein in RGCs at 12,24,and 48h after H_(2)O_(2)stimulation and in the retina at 12h after optic nerve transection(ONT).RESULTS:The applications of 5 and 10μmol/L of ET significantly increased the viability of RGCs.Results from qRT-PCR indicated a decrease in the expression of iNOS and an increase in the expressions of Nrf2 and HO-1 in ETpretreated RGCs at 12,24 and 48h after H_(2)O_(2)stimulation,as well as in ET-treated retinas at 12h after ONT.Western blot analysis revealed a decrease in the expression of iNOS and levels of conjugated acrolein,along with an increase in the expressions of Nrf2 and HO-1 in ET-pretreated RGCs in vitro and ET-treated retinas in vivo.CONCLUSION:ET is a neuroprotective agent in primary cultured RGCs injured by H_(2)O_(2).The effect of ET is dosedependent with the greatest effect being at 10μmol/L.ET plays an antioxidant role by inhibiting iNOS,up-regulating Nrf2/HO-1,decreasing the production of acrolein,and increasing the scavenge of acrolein.

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.

Effects of α-zinc sulfate combined with yiqiyangyinghuoxue therapy on related factors in patients with type 2 diabetes peripheral neuropathy

Objective: To study the effects of -zinc sulfate combined with yiqiyangyinghuoxue therapy on related factors in patients with type 2 diabetes peripheral neuropathy. Methods: A total of 90 patients with type 2 diabetes peripheral neuropathy in our hospital from September 2015 to September 2018 were enrolled in this study. The subjects were divided into the control group (n=45) and the treatment group (n=45) randomly. The control group were treated with -zinc sulfate, the treatment group were treated with -zinc sulfate combined with yiqiyangyinghuoxue therapy, the two groups were treated for 3 months. The serum NSE, UA, Hcy, hs-CRP, BDNF, HMGB1, CysC, TGF-β1, 25-(OH)D3, ESM-1, NO and plasma ET, TNF of the two groups before and after treatment were compared. Results: There were no significantly differences of the serum NSE, UA, Hcy, hs-CRP, BDNF, HMGB1, CysC, TGF-β1, 25-(OH)D3, ESM-1, NO and plasma ET, TNF of the two groups before treatment. After treatment, the serum NSE, UA, Hcy, hs-CRP, HMGB1, CysC, TGF-β1, ESM-1 and plasma ET, TNF of the two groups were significantly lower than before treatment, the serum BDNF, 25-(OH)D3, NO of the two groups were significantly higher than before treatment, and that of the treatment group after treatment were significantly better than the control group. Conclusion: α-zinc sulfate combined with yiqiyangyinghuoxue therapy on patients with type 2 diabetes peripheral neuropathy has a good efficacy, can improve the neuropathy and vascular endothelial damage, improve related factors, and it was worthy clinical application.

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。

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。

BMAL1减轻H_(2)O_(2)诱导的心肌细胞损伤机制研究

目的探讨脑和肌肉组织芳香烃受体核转运蛋白的类似蛋白1(BMAL1)通过核因子E2相关因子2(NRF2)调节活性氧(ROS)/NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体通路对过氧化氢(H_(2)O_(2))诱导的心肌细胞损伤的影响。方法体外培养H9c2细胞和BMAL1稳定过表达的H9c2细胞,建立H_(2)O_(2)诱导的H9c2细胞损伤模型,并将细胞分为对照(Control)组、H_(2)O_(2)组、BMAL1过表达(BMAL1-OE)组、BMAL1过表达+H_(2)O_(2)(BMAL1-OE+H_(2)O_(2))组、BMAL1过表达+NRF2抑制剂(BMAL1-OE+ML385)组、BMAL1过表达+NRF2抑制剂+H_(2)O_(2)(BMAL1-OE+ML385+H_(2)O_(2))组。采用CCK-8法检测细胞活力,荧光探针2’,7’-二氯荧光素二乙酸酯检测ROS生成,Western blot检测BMAL1、NRF2和NLRP3蛋白表达,酶联免疫吸附试验法检测白细胞介素(IL)-1β释放。结果与Control组相比,H_(2)O_(2)组H9c2心肌细胞活力减弱,ROS生成增多,BMAL1和NRF2蛋白表达水平降低,NLRP3蛋白表达水平升高,IL-1β释放增多(P<0.05);与H_(2)O_(2)组相比,BMAL1-OE+H_(2)O_(2)组H9c2心肌细胞活力升高,ROS生成减少,BMAL1和NRF2蛋白表达水平升高,NLRP3蛋白表达水平降低,IL-1β释放减少(P<0.05)。与BMAL1-OE+H_(2)O_(2)组相比,BMAL1-OE+ML385+H_(2)O_(2)组H9c2心肌细胞活力减弱,ROS生成增多,NLRP3蛋白表达水平升高,IL-1β释放增多(P<0.05)。结论BMAL1可减轻H_(2)O_(2)诱导的H9c2心肌细胞损伤,其机制可能与NRF2调节ROS/NLRP3炎症小体通路有关。

槲皮素预处理ALI大鼠肺组织损伤、炎症/氧化应激反应、铁死亡及Nrf2/HO-1信号通路激活情况观察

目的观察槲皮素灌胃预处理的LPS诱导急性肺损伤(ALI)大鼠肺组织损伤、炎症反应、氧化应激反应、铁死亡及Nrf2/HO-1信号通路激活情况,以探讨槲皮素对ALI的预防作用及机制。方法24只SD大鼠分为槲皮素低、中、高剂量组和阳性对照组(地塞米松)、模型组、正常对照组。槲皮素低、中、高剂量组以25、50、100 mg/kg槲皮素灌胃,1次/天,连续7 d;槲皮素灌胃处理的第4天在大鼠气管内滴注5 mg/kg LPS;阳性对照组以地塞米松1.04 mg/kg灌胃,其余处理同槲皮素组;模型组以生理盐水灌胃,1次/天,连续7天,其余处理同槲皮素组;正常对照组以生理盐水连续灌胃7 d。末次灌注给药后,观察各组肺组织损伤(肺功能及肺组织病理改变、纤维组织阳性表达率、肺泡上皮细胞凋亡率)、炎症反应(肺泡灌洗液TNF-α、IL-6、IL-1β)、氧化应激反应(肺泡灌洗液SOD、GSH、MDA,肺组织ROS)、铁死亡(Fe^(2+)水平)及Nrf2/HO-1信号通路激活[肺组织核因子红细胞2相关因子2(Nfr2)、血红素加氧酶1(HO-1)、超氧化物歧化酶2(SOD2)、过氧化氢酶(CAT)蛋白]情况。结果与正常对照组比较,模型组PaCO_(2)水平升高,PaO_(2)、SaO_(2)水平降低(P均<0.01);肺组织可见肺泡上皮细胞变性坏死,纤维组织阳性表达率高;肺泡上皮细胞凋亡率高;肺泡灌洗液中TNF-α、IL-6、IL-1β水平均升高,SOD、GSH水平降低,MDA水平升高,肺组织ROS表达水平升高,肺泡灌洗液中Fe^(2+)水平升高(P均<0.05)。与模型组相比,槲皮素中、高剂量组和阳性对照组中PaCO_(2)均降低(P均<0.05),槲皮素高剂量组PaO_(2)和SaO_(2)水平升高(P均<0.05);槲皮素高剂量组与阳性对照组肺组织炎性浸润与纤维增生明显减少,肺泡恢复正常生理结构;槲皮素低、中、高剂量组和阳性对照组肺纤维组织阳性表达率均下降(P均<0.05),其中槲皮素高剂量组肺纤维组织阳性表达率最低;槲皮素低、中、高剂量组和阳性对照组肺泡上皮细胞凋亡率均降低(P均<0.01);槲皮素低、中、高剂量组和阳性对照组肺泡灌洗液中TNF-α、IL-6、IL-1β水平均降低、SOD、GSH水平升高、MDA水平降低,肺组织ROS表达水平降低(P均<0.01),肺泡灌洗液中Fe^(2+)水平降低。与正常对照组比较,模型组肺组织中CAT、HO-1、Nrf2、SOD2蛋白表达水平低(P均<0.01);与模型组比较,槲皮素中、高剂量组和阳性对照组肺组织中CAT、HO-1、Nrf2、SOD2蛋白表达水平高(P均<0.05)。结论槲皮素灌胃预处理的ALI大鼠肺组织损伤、炎症反应、氧化应激反应、铁死亡情况减轻,Nfr2/HO-1信号通路激活;槲皮素灌胃预处理可预防LPS诱导的大鼠ALI,可能通过抑制炎症反应、氧化应激反应及铁死亡而起作用;槲皮素可能通过上调Nfr2/HO-1信号通路而抑制炎症反应、氧化应激反应及铁死亡;50、100 mg/kg槲皮素均对LPS诱导的大鼠ALI起预防作用,以100 mg/kg槲皮素的作用效果更佳。

常压高浓度氧对新生大鼠脑微血管内皮细胞损伤与Nrf2/HO-1信号通路的影响分析

目的探究常压高浓度氧(NBO)对新生大鼠脑微血管内皮细胞损伤及核因子E2相关因子2/血红素氧合酶-1(Nrf2/HO-1)信号通路的影响。方法取新生SD大鼠45只,采用随机数字表法分为常氧组、NBO组和NBO+Nrf2激活剂组,每组各15只。常氧组大鼠置于普通空气(21%氧气)中饲养,NBO组和NBO+Nrf2激活剂组大鼠置于90%常压氧气饲养,NBO+Nrf2激活剂组每日灌胃5 mg/kg Nrf2激动剂莱菔硫烷。测定脑组织伊文思蓝(EB)含量,采用酶联免疫法检测血管内皮生长因子(VEGF)和基质金属蛋白酶9(MMP-9)含量,干湿重法检测脑组织含水量,HE染色和TUNEL染色观察脑组织病理变化,蛋白质印迹法检测海马组织Nrf2/HO-1信号通路蛋白表达,水迷宫检测大鼠认知功能。结果与常氧组比较,NBO组脑组织EB、VEGF、MMP-9含量及脑组织含水量升高,差异均有统计学意义(P<0.05);与NBO组比较,NBO+Nrf2激活剂组脑组织EB、VEGF、MMP-9含量及脑组织含水量降低,差异均有统计学意义(P<0.05)。病理染色结果显示,常氧组大鼠神经细胞形态及结构完整,未见明显病理变化和细胞凋亡;NBO组神经细胞形态及结构不规则,出现明显的水肿和空泡,并伴有大量的凋亡细胞;NBO+Nrf2激活剂组脑组织病理损伤较NBO组明显减轻。与常氧组比较,NBO组脑组织Nrf2、HO-1蛋白相对表达量降低,差异均有统计学意义(P<0.05);与NBO组比较,NBO+Nrf2激活剂组Nrf2、HO-1蛋白相对表达量升高,差异均有统计学意义(P<0.05)。与常氧组比较,NBO组第2~4天逃避潜伏期延长,穿越平台次数减少,差异均有统计学意义(P<0.05);与NBO组比较,NBO+Nrf2激活剂组第2~4天逃避潜伏期缩短,穿越平台次数增多,差异均有统计学意义(P<0.05)。结论NBO可诱导新生大鼠脑微血管内皮细胞损伤,导致远期认知功能障碍,可能与下调Nrf2/HO-1信号通路表达有关。

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。