Mechanism of hesperidin improving myocardial ischemia/reperfusion injury in type 2 diabetic rats through SIRT1/Nrf2/HO-1 signaling pathway

Objective:To observe the protective effect of hesperidin on myocardial ischemia/reperfusion injury in type 2 diabetes mellitus and its effect on SIRT1/Nrf2/HO-1 signaling pathway.Methods:50 Sprague-Dawley(SD)rats were randomly assigned to the normal control group(NC),model group,ischemia-reperfusion group(IR),hesperidin group,SIRT1 inhibitor group and hesperidin plus SIRT1 inhibitor group.In addition to NC,the rats in the remaining groups were replicated by intraperitoneal of high-fat diet combined with injection of streptozotocin for type 2 diabetic rats.After then,the myocardial ischemia/reperfusion injury(MIRI)rat model was established by LAd for 30 minutes with 2 hours reperfusion.He staining was used to observe the pathological changes of myocardial tissue,and the levels of serum LDH,CK-MB and SOD,GSH and MDA in myocardial tissue were detected by kit methods,and the expression abundance of related proteins in 4-HNE and SIRT1/Nrf2/HO-1 signal pathway were detected by immunohistochemistry and Western blot;Results:Hesperidin could significantly inhibit cardiomyocyte necrosis and inflammatory cell infiltration,reduce LDH activity,CK-MB and MDA level,and increase SOD activity,GSH and 4-HNE level,the differences were statistically significant when compared with IR group(P<0.01).In addition,compared with the ischemia-reperfusion group,the expressions of SIRT1,Nrf2 and HO-1 proteins in hesperidin group were significantly up-regulated,the differences were statistically significant(P<0.01);Conclusion:Hesperidin inhibits oxidative stress by activating SIRT1/Nrf2/HO-1 signaling pathway,and play a protective effect of myocardial ischemia reperfusion injury in diabetic rats.

Effects of nuciferine on Nrf2/HO-1 signaling pathway in adipose tissue of obesity model rats

Objective:This study aimed to explore the therapeutic effect of nuciferine on high-fat diet-induced obesity in rats and the influence of nuciferine on nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway in the adipose tissue.Methods:A total of 40 male Sprague Dawley(SD)rats were evenly divided into the normal,model,positive control,and nuciferine groups,using the random number table method.Except for the normal group,rats in the other groups were fed with high-fat diet for 12 weeks to establish the obesity model.During the model establishment,rats in the positive control group received atorvastatin calcium 2 mg/kg,rats in the nuciferine group received nuciferine 20 mg/kg,and rats in the normal and model groups received normal saline 2 mL,daily through intragastric administration for 12 consecutive weeks.After model establishment and administration,the body weight,Lee’s index,and blood lipids of rats in each group were measured,and hematoxylin and eosin(HE)staining was performed on the liver and adipose tissues to evaluate the therapeutic effect of nuciferine on obesity rat model.Additionally,the levels of superoxide dismutase(SOD),malondialdehyde(MDA),and glutathione peroxidase(GSH-Px)in the serum of rats in each group were determined,and the gene expressions of Nrf2 and HO-1 in the adipose tissue of rats in each group were detected through quantitative polymerase chain reaction(qPCR)to investigate the mechanism of action of nuciferine in the treatment of obesity.Results:After 12 weeks of model establishment and administration,we observed that compared with the model group,nuciferine could significantly reduce the body weight,Lee’s index,and serum triglyceride(TG),total cholesterol(TC),and low-density lipoprotein cholesterol(LDL-C)levels and increase the serum high-density lipoprotein cholesterol(HDL-C)level in obesity rat model(P<0.05 or P<0.01).HE staining revealed that nuciferine could significantly alleviate liver steatosis in obesity rat model and improve the cell morphology in epididymal adipose tissue.Moreover,nuciferine could elevate serum SOD and GSH-Px activities in obesity rat model and lower the serum MDA level(P<0.05 or P<0.01).The qPCR indicated that nuciferine could upregulate the gene expression of Nrf2 and HO-1 in the adipose tissue of obesity rat model(P<0.05 or P<0.01).

Therapeutic Role of Chinese Medicine Targeting Nrf2/HO-1 Signaling Pathway in Myocardial Ischemia/Reperfusion Injury

Acute myocardial infarction(AMI),characterized by high incidence and mortality rates,poses a significant public health threat.Reperfusion therapy,though the preferred treatment for AMI,often exacerbates cardiac damage,leading to myocardial ischemia/reperfusion injury(MI/RI).Consequently,the development of strategies to reduce MI/RI is an urgent priority in cardiovascular therapy.Chinese medicine,recognized for its multi-component,multi-pathway,and multi-target capabilities,provides a novel approach for alleviating MI/RI.A key area of interest is the nuclear factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)pathway.This pathway is instrumental in regulating inflammatory responses,oxidative stress,apoptosis,endoplasmic reticulum stress,and ferroptosis in MI/RI.This paper presents a comprehensive overview of the Nrf2/HO-1 signaling pathway's structure and its influence on MI/RI.Additionally,it reviews the latest research on leveraging Chinese medicine to modulate the Nrf2/HO-1 pathway in MI/RI treatment.

Enhancement of porcine in vitro embryonic development through luteolin‑mediated activation of the Nrf2/Keap1 signaling pathway

Background Oxidative stress,caused by an imbalance in the production and elimination of intracellular reactive oxygen species(ROS),has been recognized for its detrimental effects on mammalian embryonic development.Luteolin(Lut)has been documented for its protective effects against oxidative stress in various studies.However,its specific role in embryonic development remains unexplored.This study aims to investigate the influence of Lut on porcine embryonic development and to elucidate the underlying mechanism.Results After undergoing parthenogenetic activation(PA)or in vitro fertilization,embryos supplemented with 0.5μmol/L Lut displayed a significant enhancement in cleavage and blastocyst formation rates,with an increase in total cell numbers and a decrease in the apoptosis rate compared to the control.Measurements on D2 and D6 revealed that embryos with Lut supplementation had lower ROS levels and higher glutathione levels compared to the control.Moreover,Lut supplementation significantly augmented mitochondrial content and membrane potential.Intriguingly,activation of the Nrf2/Keap1 signaling pathway was observed in embryos supplemented with Lut,leading to the upregulation of antioxidant-related gene transcription levels.To further validate the relationship between the Nrf2/Keap1 signaling pathway and effects of Lut in porcine embryonic development,we cultured PA embryos in a medium supplemented with brusatol,with or without the inclusion of Lut.The positive effects of Lut on developmental competence were negated by brusatol treatment.Conclusions Our findings indicate that Lut-mediated activation of the Nrf2/Keap1 signaling pathway contributes to the enhanced production of porcine embryos with high developmental competence,and offers insight into the mechanisms regulating early embryonic development.

Oleuropein alleviates sepsis-induced acute lung injury via the AMPK/Nrf-2/HO-1 signaling

Objective:To explore the effect of oleuropein on sepsis-induced acute lung injury(ALI)in vitro and in vivo and investigate the underlying mechanism.Methods:In an lipopolysaccharide(LPS)-mediated cell model of sepsis-induced ALI and a cecal ligation and puncture-induced mouse model of septic ALI,CCK-8 assay and flow cytometry analysis were used to detect cell activity and apoptosis.ELISA and relevant assay kits were used to measure the levels of inflammatory cytokines and oxidative stress,respectively.Western blot was applied to determine the expression of apoptosis-and AMP-activated protein kinase(AMPK)/nuclear factor erythroid 2-related factor-2(Nrf-2)/heme oxygenase-1(HO-1)signaling-associated proteins.JC-1 staining,adenosine triphosphate(ATP)assay kit,and MitoSOX Red assays were performed to detect mitochondrial membrane potential,ATP content,and mitochondrial ROS formation,respectively.Moreover,lung injury was evaluated by measuring lung morphological alternations,lung wet-to-dry ratio,myeloperoxidase content,and total protein concentration.Results:Oleuropein reduced inflammatory reaction,oxidative damage,and apoptosis,and ameliorated mitochondrial dysfunction in LPS-exposed BEAS-2B cells and mice with septic ALI.Besides,oleuropein activated the AMPK/Nrf-2/HO-1 signaling pathway.However,these effects of oleuropein were abrogated by an AMPK inhibitor compound C.Conclusions:Oleuropein can protect against sepsis-induced ALI in vitro and in vivo by activating the AMPK/Nrf-2/HO-1 signaling,which might be a potential therapeutic agent for the treatment of sepsis-induced ALI.

X-Paste improves wound healing in diabetes via NF-E2-related factor/HO-1 signaling pathway

BACKGROUND Diabetic foot ulcers(DFU),as severe complications of diabetes mellitus(DM),significantly compromise patient health and carry risks of amputation and mortality.AIM To offer new insights into the occurrence and development of DFU,focusing on the therapeutic mechanisms of X-Paste(XP)of wound healing in diabetic mice.METHODS Employing traditional Chinese medicine ointment preparation methods,XP combines various medicinal ingredients.High-performance liquid chromatography(HPLC)identified XP’s main components.Using streptozotocin(STZ)-induced diabetic,we aimed to investigate whether XP participated in the process of diabetic wound healing.RNA-sequencing analyzed gene expression differences between XP-treated and control groups.Molecular docking clarified XP’s treatment mechanisms for diabetic wound healing.Human umbilical vein endothelial cells(HUVECs)were used to investigate the effects of Andrographolide(Andro)on cell viability,reactive oxygen species generation,apoptosis,proliferation,and metastasis in vitro following exposure to high glucose(HG),while NF-E2-related factor-2(Nrf2)knockdown elucidated Andro’s molecular mechanisms.RESULTS XP notably enhanced wound healing in mice,expediting the healing process.RNA-sequencing revealed Nrf2 upregulation in DM tissues following XP treatment.HPLC identified 21 primary XP components,with Andro exhibiting strong Nrf2 binding.Andro mitigated HG-induced HUVECs proliferation,metastasis,angiogenic injury,and inflammation inhibition.Andro alleviates HG-induced HUVECs damage through Nrf2/HO-1 pathway activation,with Nrf2 knockdown reducing Andro’s proliferative and endothelial protective effects.CONCLUSION XP significantly promotes wound healing in STZ-induced diabetic models.As XP’s key component,Andro activates the Nrf2/HO-1 signaling pathway,enhancing cell proliferation,tubule formation,and inflammation reduction.

Hesperidin ameliorates H_(2)O_(2)-induced bovine mammary epithelial cell oxidative stress via the Nrf2 signaling pathway

Background Hesperidin is a citrus flavonoid with anti-inflammatory and antioxidant potential. However, its protective effects on bovine mammary epithelial cells(b MECs) exposed to oxidative stress have not been elucidated.Results In this study, we investigated the effects of hesperidin on H_(2)O_(2)-induced oxidative stress in b MECs and the underlying molecular mechanism. We found that hesperidin attenuated H_(2)O_(2)-induced cell damage by reducing reactive oxygen species(ROS) and malondialdehyde(MDA) levels, increasing catalase(CAT) activity, and improving cell proliferation and mitochondrial membrane potential. Moreover, hesperidin activated the Keap1/Nrf2/ARE signaling pathway by inducing the nuclear translocation of Nrf2 and the expression of its downstream genes NQO1 and HO-1, which are antioxidant enzymes involved in ROS scavenging and cellular redox balance. The protective effects of hesperidin were blocked by the Nrf2 inhibitor ML385, indicating that they were Nrf2 dependent.Conclusions Our results suggest that hesperidin could protect b MECs from oxidative stress injury by activating the Nrf2 signaling pathway, suggesting that hesperidin as a natural antioxidant has positive potential as a feed additive or plant drug to promote the health benefits of bovine mammary.

Flavone improves liver damage in nicotine-exposed rats via the Nrf2/HO-1 pathway

Objective:To assess the hepatoprotective effects of flavone on nicotine-induced liver damage.Methods:Thirty-six rats were allocated into six groups:the control group,the nicotine group,the flavone alone groups(10 and 25 mg/kg/body weight),and the nicotine groups treated with flavone(10 and 25 mg/kg/body weight).Liver function,oxidative stress,Nrf2 pathway(HO-1,Nrf2,and Keap-1),and inflammatory markers(IL-17,TNF-α,and NF-κB)were evaluated.Additionally,a histopathological examination of liver tissues was performed.Results:Nicotine increased liver damage,inflammation,and oxidative stress.However,flavone suppressed nicotine-induced liver enzymes,oxidative stress,and inflammation,as manifested by increased antioxidants and decreased malondialdehyde level,liver enzymatic activities,and inflammatory markers.Flavone(10 and 25 mg/kg/body weight)also reduced the level of Keap-1 and increased HO-1 and Nrf2 levels in the liver of nicotine-exposed rats.Conclusions:Flavone has hepatoprotective properties and may slow the progression of liver injury by reducing oxidative stress,liver enzymes,and inflammation possibly via the Nrf2 pathway.

Exploring the mechanism of electroacupuncture at different acupoints on acute colitis rats based on JAK2/STAT3/SOCS1 signaling pathway

Objective:To investigate the mechanism of JAK2/STAT3/SOCS1 signaling pathway in electroacupuncture of different acupoints on acute colitis rats.Methods:36 SPF SD rats were randomly divided into 6 groups,with 6 rats in each group.The rat model of acute colitis was prepared by enema with glacial acetic acid solution.After the model was established,electroacupuncture was given to each acupoint group,with density wave,frequency 2Hz-50 Hz,intensity 2 mA,muscle tremor as the degree 20 min/time,1 time/day,for 3 consecutive days.Observe the general condition of rats;the pathological changes of colonic mucosa in rats were observed by HE method.The contents of serum interleukin-4(IL-4)and interleukin-8(IL-8)were detected by ELISA.Western blot and RT-PCR were used to detect the expression of JAK2,STAT3,SOCS1 protein and mRNA in rat colon tissue.Results:In contrast to the normal group,the overall condition of the model group was worse,the colonic mucosa was severely damaged,even necrotic,and the ulcer surface was obvious.The content of IL-4 in serum was obviously reduced,and the content of IL-8 was obviously go up(P<0.01).The protein content of JAK2,STAT3 and the expression of JAK2,STAT3 mRNA in colon tissue of rats were obviously go up,while the protein content of SOCS1 and the expression of SOCS1 mRNA were obviously reduced(P<0.01).In contrast to the model group,the general condition of rats in each acupoint group was significantly improved,the damage and necrosis of colonic mucosa and ulcer surface were obviously alleviated,the content of IL-4 in serum was obviously go up,and the content of IL-8 was significantly decreased(P<0.01).The protein content of JAK2,STAT3 and the expression of JAK2,STAT3 mRNA in colon tissue of rats were obviously reduced,while the protein content of SOCS1 and the expression of SOCS1 mRNA were obviously go up(P<0.05,P<0.01).Comparison of different acupoint groups,the colonic mucosal injury in the Zusanli group was significantly reduced,the content of serum IL-4 was significantly increased,and the content of IL-8 was significantly decreased(P<0.05,P<0.01).The protein content and mRNA expression of JAK2 and STAT3 in colon tissue were significantly down-regulated,while the protein content and mRNA expression of SOCS1 were significantly go up(P<0.05,P<0.01).Conclusion:Electroacupuncture at each acupoint can improve the damage of colonic mucosa and reduce the inflammatory response.The therapeutic effect of Zusanli(ST36)is better than that of Tianshu(ST25),Dachangshu(BL25)and Shangjuxu(ST37).The mechanism may be related to the regulation of JAK2/STAT3/SOCS1 signaling pathway related proteins and inflammatory cytokines IL-4 and IL-8.

枸杞多糖调节Nrf2/HO-1/GPX4铁死亡途径对妊娠期糖尿病大鼠胰岛素抵抗的改善作用

目的 探讨枸杞多糖调节Nrf2/HO-1/GPX4铁死亡途径对妊娠期糖尿病(GDM)大鼠胰岛素抵抗的改善作用。方法 将SD成年大鼠雌雄合笼得到孕鼠,建立GDM模型,随机分为模型组,枸杞多糖低、中、高剂量组,ferrostatin-1组,每组8只,另设对照组,腹腔注射等剂量柠檬酸缓冲液。末次给药24 h后,检测大鼠FBG、FINS、IRI水平。再次复制GDM模型32只,随机分为模型组、枸杞多糖组、ML385组、枸杞多糖+ML385组,另设对照组。药物分组干预后,检测大鼠FBG、FINS、IRI、TG、TC水平,妊娠第19天检测母体及胎鼠体质量以及血清IL-18、IL-6、SOD、MDA水平,子宫内膜组织ACSL4、PTGS2、Nrf2/HO-1/GPX4通路蛋白表达。结果 与对照组比较,模型组大鼠FBG、FINS、IRI,母体及胎鼠体质量,血清TG、TC、IL-18、IL-6、MDA水平,子宫内膜组织ACSL4、PTGS2蛋白表达升高(P<0.05),血清SOD水平、子宫内膜组织Nrf2、HO-1、GPX4蛋白表达降低(P<0.05);与枸杞多糖+ML385组比较,枸杞多糖组大鼠FBG、FINS、IRI,母体及胎鼠体质量,血清TG、TC、IL-18、IL-6、MDA水平,子宫内膜组织ACSL4、PTGS2蛋白表达降低(P<0.05),血清SOD、子宫内膜组织Nrf2、HO-1、GPX4蛋白表达升高(P<0.05)。结论 枸杞多糖可通过促进Nrf2/HO-1/GPX4信号传导,抑制铁死亡途径,降低GDM大鼠血糖血脂水平,减弱其胰岛素抵抗。

枸杞多糖通过激活Nrf2/HO-1通路保护HK-2细胞氧化损伤的作用

目的分析枸杞多糖(LBP)干预对氧化损伤的人肾小管上皮细胞(HK-2)内的核因子E2相关因子2(Nrf2)/血红素氧合酶-1(HO-1)通路蛋白表达的变化,探索LBP拮抗HK-2细胞氧化损伤的分子机制。方法采用过氧化氢诱导HK-2细胞制备氧化损伤细胞模型,HK-2被分成4组:正常组、LBP组、氧化损伤组及LBP干预组。观察4组细胞的长势和形态变化;细胞活力测定法比较每组细胞的存活率;比色法分析氧化产物丙二醛(malondialdehyde,MDA)含量以及超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化酶(GSH-Px)的水平;免疫印迹技术检测细胞内Nrf2/HO-1通路蛋白Nrf2、HO-1的相对水平。结果正常组和LBP组相比,两组的细胞长势、形态和存活率,细胞产生MDA的量、SOD和GSH-Px的水平,通路蛋白Nrf2、HO-1的相对水平均无明显差异(均P>0.05);氧化损伤组和正常组相比,细胞皱缩变圆并脱落、贴壁细胞变少,存活率减少、MDA量增加、SOD和GSH-Px降低,通路蛋白Nrf2、HO-1的相对值下降(均P<0.05);LBP干预组和氧化损伤组相比,细胞长势、形状恢复、贴壁细胞较多,存活率增加、MDA量减少、SOD和GSH-Px升高,通路蛋白Nrf2、HO-1的相对值升高(均P<0.05)。结论枸杞多糖能通过激活Nrf2/HO-1通路提高HK-2细胞的抗氧化酶活性达到减轻细胞氧化损伤的目的。

有氧运动通过Keap1Nrf2HO-1通路对睡眠障碍老年大鼠认知的改善机制

目的探究有氧运动通过Keap1Nrf2HO-1通路对睡眠障碍老年大鼠认知的改善机制。方法选用Wistar大鼠,60只,雌雄各半,将大鼠均分为4组,其中一组为空白组,其余分为模型组、给药组(根据每只大鼠的体重按照0.5 mg/kg进行艾司唑仑片灌胃给药)和有氧运动组,每组15只。结果与空白组比较,模型组逃避潜伏期延长(P<0.01),且伴随着明显的寻台次数较少、穿越原平台次数降低(P<0.01),大鼠在原平台停留的时间占总游泳时间的百分比降低(P<0.05);与模型组比较,给药组和有氧运动组逃避潜伏期缩短(P<0.01),且伴随着明显的寻台次数增加、穿越原平台次数增加(P<0.01),在原平台停留的时间占总游泳时间的百分比升高(P<0.05)。与空白组比较,模型组MDA水平升高,SOD活性降低(P<0.01),Keap1水平升高(P<0.01),Nrf2和HO-1水平降低(P<0.01);与模型组比较,给药组和有氧运动组MDA水平降低,SOD活性升高(P<0.01),Keap1水平降低,Nrf2和HO-1水平升高(P<0.01)。结论有氧运动能够有效缓解失眠障碍大鼠的认知功能,其机制原理可能与有氧运动通过改善睡眠障碍大鼠机体内海马组织的Keap1/Nrf2/HO-1蛋白表达水平从而产生抗氧化应急作用有关。

西黄胶囊联合西妥昔单抗通过Nrf2/HO-1信号通路调控结直肠癌细胞凋亡、迁移以及侵袭的作用机制研究

目的研究西黄胶囊联合西妥昔单抗通过Nrf2/HO-1信号通路对结直肠癌细胞凋亡、迁移以及侵袭的调控作用。方法使用细胞计数试剂盒(CCK8)确定西黄胶囊和西妥昔单抗对结直肠癌HCT-116细胞活力的影响。使用流式细胞术、划痕法和侵袭小室法检测西黄胶囊联合西妥昔单抗对结直肠癌HCT-116细胞凋亡、迁移以及侵袭的影响。使用蛋白质印迹技术检测西黄胶囊联合西妥昔单抗对结直肠癌HCT-116细胞B细胞淋巴瘤2(BCL2)、BCL2关联X蛋白(BAX)、基质金属蛋白酶2(MMP2)、基质金属蛋白酶9(MMP9)、锌指蛋白232(ZNF320)、血管内皮生长因子A(VEGA)、糖原合成酶激酶3β(GSK3β)、核因子E2相关因子2(NRF2)和血红素氧合酶1(HO-1)表达的影响。结果CCK8实验结果显示,西黄胶囊和西妥昔单抗对HCT-116细胞活力最佳抑制时间是72 h,IC 50分别为5.28%和170.20 mg/L。与对照组相比,西黄胶囊和西妥昔单抗增加HCT-116细胞的凋亡(P<0.05)、抑制HCT-116细胞迁移以及侵袭(P<0.05)。与单独使用西黄胶囊或西妥昔单抗相比,西黄胶囊联合西妥昔单抗对HCT-116细胞的促凋亡效果和对迁移以及侵袭的抑制效果更加明显(P<0.05)。蛋白质印迹技术结果显示,与对照组比较,西黄胶囊和西妥昔单抗促进HCT-116细胞的BAX表达(P<0.05),抑制BCL2、MMP2、MMP9、ZNF320、VEGA、GSK3B、NRF2、HO-1的表达(P<0.05)。此外,与单独使用西黄胶囊或西妥昔单抗相比,西黄胶囊联合西妥昔单抗的效果更加明显(P<0.05)。结论西黄胶囊联合西妥昔单抗可能通过抑制Nrf2/HO-1信号通路抑制结直肠癌HCT-116细胞的活力、迁移以及侵袭,促进细胞凋亡,且西黄胶囊联合西妥昔单抗的干预效果优于单独使用西黄胶囊或西妥昔单抗。

金雀异黄酮通过Nrf2/HO-1信号通路减轻皮质神经元低氧/复氧损伤

目的研究金雀异黄酮(GEN)对皮质神经元低氧/复氧(H/R)损伤的影响,并基于核因子E2相关因子2/血红素加氧酶1(Nrf2/HO-1)信号通路探讨其机制。方法分离并体外培养C57BL/6J小鼠胎鼠(妊娠15 d)皮质神经元,设正常对照(Normal)组、模型(H/R)组、GEN(12.5μmol·L^(-1))组、TBHQ(Nrf2激动剂,10μmol·L^(-1))组、GEN(12.5μmol·L^(-1))+TBHQ(10μmol·L^(-1))组。除正常对照组外,其他组采用低氧(5%CO_(2)+95%N_(2))4 h后复氧(5%CO_(2)+95%空气)24 h的方法制备H/R损伤皮质神经元模型,各组分别于造模前2h给药干预。采用CCK-8法、流式细胞术检测神经元活力和凋亡率,DCFH-DA荧光探针法检测神经元活性氧(ROS)含量,分光光度法检测神经元中丙二醛(MDA)含量和抗氧化酶[超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)]活性,RT-PCR法检测神经元Nrf2、HO-1 mRNA表达,Western blot法检测神经元Nrf2、HO-1、B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)、激活型Caspase-3(cleaved Caspase-3)蛋白表达。结果与H/R组比较,GEN组、TBHQ组和GEN+TBHQ组皮质神经元活力明显升高、凋亡率明显降低(P<0.05);神经元中ROS、MDA含量明显降低,SOD、GSH-Px活性明显升高(P<0.05);Nrf2、HO-1 mRNA表达量明显升高(P<0.05);Nrf2、HO-1、Bcl-2蛋白表达量及Bcl-2/Bax比值明显升高,Bax、cleaved Caspase-3蛋白表达量明显降低(P<0.05)。GEN+TBHQ组对H/R损伤皮质神经元活力、凋亡率、氧化应激指标、Nrf2/HO-1信号通路相关mRNA和蛋白表达的调控作用均明显优于GEN组和TBHQ组(P<0.05)。结论GEN可通过促进Nrf2/HO-1信号通路活化抑制氧化应激损伤和神经元凋亡,对皮质神经元H/R损伤起到保护作用。

梓醇调节Keap1/Nrf2/HO-1信号通路对口腔鳞癌细胞恶性生物学行为的影响

目的探究梓醇调节Kelch样环氧氯丙烷相关蛋白-1/核因子E2相关因子2/血红素氧合酶-1(Keap1/Nrf2/HO-1)信号通路对口腔鳞癌细胞恶性生物学行为的影响。方法体外培养口腔鳞癌细胞Tac8113;CCK-8法筛选梓醇最佳作用水平;将Tac8113细胞分为对照组、梓醇组(24μg/mL)、sh-NC组(转染sh-NC慢病毒质粒)、sh-Keap1组(转染sh-Keap1慢病毒质粒)、梓醇+sh-NC组(转染sh-NC+24μg/mL梓醇)、梓醇+sh-Keap1组(转染sh-Keap1+24μg/mL梓醇);CCK-8法检测细胞增殖;划痕试验检测细胞迁移;流式细胞术检测细胞凋亡;2’,7’-二氯荧光素二乙酸酯(DCFH-DA)检测细胞活性氧(ROS)水平;采用试剂盒分别检测超氧化物歧化酶(SOD)、丙二醛(MDA)水平;Western Blot分别检测Keap1/Nrf2/HO-1信号通路及凋亡相关蛋白表达水平。结果与0μg/mL组比较,随着梓醇剂量增加Tac8113细胞存活率显著降低(P<0.05),因此,选择24μg/mL梓醇作为后续实验的干预条件;与对照组比较,梓醇组Tac8113细胞OD450值、划痕愈合率、ROS水平、MDA水平及B淋巴细胞瘤-2(Bcl-2)、Nrf2、HO-1表达显著降低,细胞凋亡率、SOD水平及Keap1、胱天蛋白酶3(caspase3)表达显著升高(P<0.05);Keap1低表达后Tac8113细胞恶性生物学行为程度加重,且逆转了梓醇对Tac8113细胞恶性生物学行为的影响。结论梓醇抑制口腔鳞癌细胞增殖、迁移,诱导细胞凋亡发挥抑癌作用,可能与上调Keap1表达,下调Nrf2和HO-1表达有关。

督脉隔药灸联合水凝胶多功能复合敷料对压力性损伤大鼠Nrf2/HO-1/NQO1信号通路的影响

目的观察督脉隔药灸联合水凝胶多功能复合敷料对压力性损伤大鼠创面修复及核因子E2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)/NADPH醌氧化还原酶1(NADPH quinone oxidoreductase 1,NQO1)信号通路的影响。方法选取50只SPF级SD大鼠,随机分为假手术组、模型组、水凝胶敷料组、督脉隔药灸组、联合干预组,每组10只。除假手术组外,其余4组借助压力装置构建压力性损伤大鼠模型。分组干预并记录各组大鼠的压力性损伤修复情况及创面组织病理变化;借助ELISA检测各组大鼠的血清炎性因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素(interleukin,IL)-1β、IL-6以及氧化应激因子丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)水平;Western blot与RT-PCR检测各组大鼠创面肉芽组织Nrf2、HO-1、NQO1、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)蛋白及mRNA的表达水平。结果HE染色结果显示:假手术组皮肤结构未被破坏;模型组创面肉芽组织可见明显的细胞坏死、变性及炎性细胞浸润;干预各组可见创面修复,表现为散在炎性细胞浸润,“气球样”坏死变性明显减少。Masson染色结果显示:假手术组创面皮肤组织胶原纤维较为整齐;模型组创面肉芽组织可见大面积的胶原纤维沉积、紊乱;干预各组可见不同程度的创面修复。与假手术组对比,模型组大鼠血清TNF-α、IL-1β、IL-6、MDA水平和创面肉芽组织Nrf2、HO-1、NQO1、MMP-9蛋白及mRNA的表达水平均升高(P<0.05),而创面修复率、血清SOD、GSH-Px水平均下降(P<0.05);与模型组比较,水凝胶敷料组、督脉隔药灸组、联合干预组的血清TNF-α、IL-1β、IL-6、MDA水平均下降(P<0.05),而创面修复率、血清SOD、GSH-Px水平和创面肉芽组织Nrf2、HO-1、NQO1、MMP-9蛋白及m RNA的表达水平均升高(P<0.05),且联合干预组较其他干预组更优(P<0.05)。结论督脉隔药灸可有效促进压力性损伤的创面修复,与水凝胶多功能复合敷料联合使用效果更佳,其机制可能与督脉隔药灸抑制创面炎性反应及氧化应激,并激活Nrf2/HO-1/NQO1信号通路相关。

白花蛇舌草通过Nrf2/HO-1通路对肺腺癌增殖、迁移和侵袭的作用机制研究

目的研究白花蛇舌草对肺腺癌A549、H1975细胞株增殖、迁移和侵袭的影响及其可能机制。方法使用CCK-8实验、划痕实验、Transwell实验检测白花蛇舌草干预后对A549、H1975细胞增殖、迁移和侵袭的影响;通过RT-qPCR和Western blot实验检测白花蛇舌草干预后各组细胞Nrf2、HO-1 mRNA及蛋白的表达。结果白花蛇舌草干预后,与对照组比较,A549、H1975细胞的增殖、迁移及侵袭能力下调(P<0.05);各组细胞Nrf2、HO-1的mRNA和蛋白表达均降低(P<0.05)。结论白花蛇舌草可抑制肺腺癌A549、H1975细胞的增殖、迁移和侵袭,且该作用可能是通过调控Nrf2/HO-1信号通路完成的。

益气活血托毒方对慢性非细菌性前列腺炎大鼠Keap1/Nrf2/HO-1信号通路的影响

目的观察益气活血托毒方对慢性非细菌性前列腺炎(CNP)大鼠Keap1/Nrf2/HO-1信号通路的影响,探讨其治疗CNP作用机制。方法采用去势结合雌激素诱导法制备CNP大鼠模型。48只SD大鼠采用随机数字表法分为空白组、模型组、塞来昔布组和益气活血托毒方组,每组12只。塞来昔布组予塞来昔布混悬液0.035 g/kg灌胃,益气活血托毒方组予益气活血托毒方水煎剂8.64 g/kg灌胃,空白组和模型组灌胃等体积生理盐水,连续28 d。Von Frey纤维丝测痛仪测定大鼠机械痛阈值,HE染色观察前列腺组织病理变化并进行病理评分,化学荧光法检测前列腺组织活性氧(ROS)含量,比色法测定前列腺组织谷胱甘肽过氧化物酶(GSH-Px)活性和丙二醛(MDA)含量,Western blot检测前列腺组织Kelch样ECH相关蛋白1(Keap1)、核因子E2相关因子2(Nrf2)、血红素氧合酶-1(HO-1)蛋白表达。结果与空白组比较,模型组大鼠机械痛阈值、前列腺指数显著降低(P<0.01);前列腺组织腺腔大小不一,腔内分泌物消失,间质疏松、水肿,有大量纤维组织增生和炎性细胞浸润,病理评分显著升高(P<0.01);前列腺组织ROS、MDA含量显著升高,GSH-Px活性显著降低(P<0.01),Keap1、Nrf2蛋白表达显著降低,HO-1蛋白表达显著升高(P<0.01,P<0.05)。与模型组比较,益气活血托毒方组大鼠机械痛阈值显著升高(P<0.01);前列腺组织轻微损伤,有少量纤维增生和炎性细胞浸润,病理评分显著降低(P<0.01,P<0.05);前列腺组织ROS、MDA含量显著降低,GSH-Px活性显著升高(P<0.01),Keap1、Nrf2蛋白表达显著升高,HO-1蛋白表达显著降低(P<0.01,P<0.05)。结论益气活血托毒方可有效改善CNP大鼠前列腺组织病理形态,提高痛阈值,其作用机制可能与激活Keap1/Nrf2/HO-1信号通路,降低大鼠前列腺组织氧化应激损伤有关。

基于Keap1/Nrf2/HO-1信号通路研究当归醇提物对多囊卵巢综合征大鼠的保护作用

目的使用来曲唑联合高脂饮食建立多囊卵巢综合征(polycystic ovarian syndrome,PCOS)大鼠模型,探讨当归醇提物(ethanol extract of Angelicae Sinensis Radix,EEA)对PCOS大鼠的保护作用及其作用机制。方法来曲唑联合高脂饮食诱导雌性SD大鼠建立PCOS模型,并用EEA干预。阴道涂片染色、HE染色、酶联免疫吸附法(ELISA)和生化指标检测评价EEA对PCOS大鼠的治疗作用,并通过荧光定量PCR(qRT-PCR)和Western blot检测EEA对Keap1/Nrf2/HO-1信号通路的影响。结果阴道涂片和HE染色结果表明,EEA改善了PCOS大鼠动情周期紊乱和卵巢组织病变,ELISA和生化指标检测结果表明EEA可以调节激素紊乱,改善血脂异常;且在模型组中Keap1蛋白和mRNA表达明显上调,Nrf2和HO-1蛋白和mRNA表达明显下调而经过EEA治疗后Keap1蛋白和mRNA表达明显下调,Nrf2和HO-1蛋白和mRNA表达明显上调(P<0.05或P<0.01)。结论EEA可以减轻大鼠PCOS,其机制可能是通过促进Keap1/Nrf2/HO-1信号通路,从而抑制氧化应激。

冰菖散通过Nrf2/HO-1信号通路改善APP/PS1阿尔茨海默病小鼠铁死亡及认知功能障碍的研究

目的探讨吸嗅冰菖散通过调节Nrf2/HO-1信号通路来改善阿尔茨海默病(AD)小鼠的铁死亡和认知功能障碍。方法30只APP/PS1小鼠随机分为模型组(APP/PS1组)、冰菖散低剂量组(BCS-L组)和冰菖散高剂量组(BCS-H组),10只同窝阴性小鼠作为空白组(WT组)。WT组和APP/PS1组进行纯水雾化给药处理;BCS-L组和BCS-H组每日雾化给药0.5、1μL冰菖散吸嗅剂,每日10 min,持续2周。给药2周后进行水迷宫实验来判断各组小鼠空间认知能力,水迷宫实验后处死小鼠,取大脑组织进行Aβ_(1-42)免疫荧光和磷酸化Tau蛋白(P-tau)免疫组化及尼氏染色;提取海马组织进行Western blot、qPCR检测Keap1、Nrf2、HO-1、GPX4、SCL7A11、TFRC、DMT1、FTL、FTH1的蛋白和mRNA表达水平;提取大脑组织进行GSH试剂盒检测。结果BCS各组逃避潜伏期缩短,平台所在象限活动时间和穿越平台次数增加。相对于APP/PS1组,BCS各组小鼠的海马CA1区和皮层中的Aβ_(1-42)和P-tau表达水平显著降低(P<0.05,P<0.01)。尼氏染色显示,BCS各组神经元排列较为紧密,有较多尼氏体。在海马组织中,相比于未处理的APP/PS1小鼠,BCS各组在Nrf2、HO-1、SLC7A11、GPX4、FTL的蛋白和mRNA表达水平上均有显著提升(P<0.05,P<0.01),BCS-L组FTH1的蛋白和mRNA水平有提升,但无显著意义,同时TFRC、DMT1、Keap1的表达水平显著下调(P<0.05,P<0.01)。此外,BCS各组能有效恢复其大脑中GSH的含量(P<0.05,P<0.01)。结论吸嗅BCS可以改善APP/PS1小鼠认知功能障碍。BCS可以激活Nrf2/HO-1通路,增加GPX4的表达,并通过抑制Keap1来增强Nrf2的转录活性,从而提高SLC7A11的表达并恢复GSH的含量,有效对抗铁死亡。同时,冰菖散还能有效抑制TFRC和DMT1,上调FTL和FTH1的表达,进而维持细胞内铁的稳态平衡,有助于减轻铁死亡对APP/PS1小鼠的影响,可改善AD小鼠的认知功能。