Background:Although the buried wood of Phoebe zhennan is known as the“mummy”of the plant kingdom,there is little research on its pharmacological activity.This study endeavored to investigate the effect and mechanism...Background:Although the buried wood of Phoebe zhennan is known as the“mummy”of the plant kingdom,there is little research on its pharmacological activity.This study endeavored to investigate the effect and mechanism of buried wood of Phoebe zhennan extract(BPE)on physical fatigue mice induced by weight-loaded forced swimming.Methods:Firstly,BPE was obtained by 70%ethanol extraction and freeze-drying processes.Then,the effect of BPE on physical fatigue mice was evaluated by swimming time,rotating stick time,levels of lipid peroxidation,lactate,lactate dehydrogenase,urea nitrogen,creatine kinase and muscle glycogen.Finally,real time fluorescence quantification and western blot were used to investigate the possible mechanism of BPE.Results:BPE could significantly alleviate muscle tissue damage,prolong the exhaustion time of weight-bearing swimming and rotating stick time.Meanwhile,BPE treatment could notably reduce the accumulation of serum lactate,urea nitrogen,and activities of lactate dehydrogenase and creatine kinase,while increasing the levels of glycogen and activities of glutathione peroxidase and superoxide dismutase in muscles.Moreover,BPE treatment obviously increased HO-1,Nrf-2,AMPK,PGC-1αmRNA and protein expressions in the muscles of physical fatigue mice.Conclusion:BPE treatment could ameliorate various impairments and oxidative stress injury induced by physical fatigue via activating Nrf-2/HO-1 and AMPK/PGC-1αsignaling pathway.展开更多
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-i...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.展开更多
目的:探讨白花丹素作为一种新型的铁死亡诱导剂在膀胱癌抑制中的作用机制。方法:本研究中使用了膀胱癌细胞T24。采用细胞增殖与活性检测-8(CCK-8)法检测白花丹素(0.1、1、2、3、6、12、24、48μmol·L^(-1))对T24细胞活力的影响。...目的:探讨白花丹素作为一种新型的铁死亡诱导剂在膀胱癌抑制中的作用机制。方法:本研究中使用了膀胱癌细胞T24。采用细胞增殖与活性检测-8(CCK-8)法检测白花丹素(0.1、1、2、3、6、12、24、48μmol·L^(-1))对T24细胞活力的影响。采用膜联蛋白V-异硫氰酸荧光素/碘化丙啶(Annexin V FITC/PI)凋亡试剂盒检测白花丹素(1.5、3、6μmol·L^(-1))对T24细胞凋亡的影响。采用不同的抑制剂(铁死亡抑制剂Fer-1,凋亡抑制剂VAD,坏死性凋亡抑制剂Nec-1)与白花丹素(6μmol·L^(-1))联合使用。采用活性氧荧光探针(DCFH-DA),丙二醛(MDA)和谷胱甘肽(GSH)试剂盒分别检测不同浓度的白花丹素(1.5、3、6μmol·L^(-1))对T24细胞内活性氧水平,MDA和GSH的含量,脂质过氧化荧光探针(C11-BODIPY)荧光探针检测白花丹素(1.5、3、6μmol·L^(-1))对T24细胞中过氧化物水平的影响。蛋白免疫印迹法(Western blot)检测白花丹素(1.5、3、6μmol·L^(-1))细胞中溶质载体家族7成员11(SLC7A11)、谷胱甘肽过氧化酶(GPX4)、核因子E2相关因子-2(Nrf-2)和Kelch样ECH关联蛋白1(Keap1)的蛋白表达的影响。结果:与空白组比较,白花丹素组T24细胞的活性明显降低(P<0.05),IC50为3.52μmol·L^(-1)。与空白组比较,白花丹素组(1.5、3、6μmol·L^(-1))T24细胞凋亡率明显升高(P<0.05);与单独使用6μmol·L^(-1)的白花丹素组比较,铁死亡抑制剂和凋亡抑制剂组能够逆转6μmol·L^(-1)的白花丹素对T24细胞增殖抑制作用(P<0.05)。与空白组比较,白花丹素组(1.5、3、6μmol·L^(-1)),T24细胞ROS、MDA及脂质过氧化物的含量明显升高,GSH水平明显降低,铁死亡相关蛋白SLC7A11、GPX4以及Nrf-2/Keap1明显降低(P<0.05)。结论:白花丹素能诱导细胞铁死亡,其机制与Nrf-2/Keap1信号通路有关。展开更多
基金supported by the Scientific Research Foundation for the introduction of talent of Pingdingshan University(No.PXY-BSQD-2022040,PXY-BSQD-2023024)Henan Province Science and Technology Research Project(No.242102310313,232102310460).
文摘Background:Although the buried wood of Phoebe zhennan is known as the“mummy”of the plant kingdom,there is little research on its pharmacological activity.This study endeavored to investigate the effect and mechanism of buried wood of Phoebe zhennan extract(BPE)on physical fatigue mice induced by weight-loaded forced swimming.Methods:Firstly,BPE was obtained by 70%ethanol extraction and freeze-drying processes.Then,the effect of BPE on physical fatigue mice was evaluated by swimming time,rotating stick time,levels of lipid peroxidation,lactate,lactate dehydrogenase,urea nitrogen,creatine kinase and muscle glycogen.Finally,real time fluorescence quantification and western blot were used to investigate the possible mechanism of BPE.Results:BPE could significantly alleviate muscle tissue damage,prolong the exhaustion time of weight-bearing swimming and rotating stick time.Meanwhile,BPE treatment could notably reduce the accumulation of serum lactate,urea nitrogen,and activities of lactate dehydrogenase and creatine kinase,while increasing the levels of glycogen and activities of glutathione peroxidase and superoxide dismutase in muscles.Moreover,BPE treatment obviously increased HO-1,Nrf-2,AMPK,PGC-1αmRNA and protein expressions in the muscles of physical fatigue mice.Conclusion:BPE treatment could ameliorate various impairments and oxidative stress injury induced by physical fatigue via activating Nrf-2/HO-1 and AMPK/PGC-1αsignaling pathway.
基金supported by Wenzhou Scientific Research Project(Y20210290).
文摘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.
文摘目的:探讨白花丹素作为一种新型的铁死亡诱导剂在膀胱癌抑制中的作用机制。方法:本研究中使用了膀胱癌细胞T24。采用细胞增殖与活性检测-8(CCK-8)法检测白花丹素(0.1、1、2、3、6、12、24、48μmol·L^(-1))对T24细胞活力的影响。采用膜联蛋白V-异硫氰酸荧光素/碘化丙啶(Annexin V FITC/PI)凋亡试剂盒检测白花丹素(1.5、3、6μmol·L^(-1))对T24细胞凋亡的影响。采用不同的抑制剂(铁死亡抑制剂Fer-1,凋亡抑制剂VAD,坏死性凋亡抑制剂Nec-1)与白花丹素(6μmol·L^(-1))联合使用。采用活性氧荧光探针(DCFH-DA),丙二醛(MDA)和谷胱甘肽(GSH)试剂盒分别检测不同浓度的白花丹素(1.5、3、6μmol·L^(-1))对T24细胞内活性氧水平,MDA和GSH的含量,脂质过氧化荧光探针(C11-BODIPY)荧光探针检测白花丹素(1.5、3、6μmol·L^(-1))对T24细胞中过氧化物水平的影响。蛋白免疫印迹法(Western blot)检测白花丹素(1.5、3、6μmol·L^(-1))细胞中溶质载体家族7成员11(SLC7A11)、谷胱甘肽过氧化酶(GPX4)、核因子E2相关因子-2(Nrf-2)和Kelch样ECH关联蛋白1(Keap1)的蛋白表达的影响。结果:与空白组比较,白花丹素组T24细胞的活性明显降低(P<0.05),IC50为3.52μmol·L^(-1)。与空白组比较,白花丹素组(1.5、3、6μmol·L^(-1))T24细胞凋亡率明显升高(P<0.05);与单独使用6μmol·L^(-1)的白花丹素组比较,铁死亡抑制剂和凋亡抑制剂组能够逆转6μmol·L^(-1)的白花丹素对T24细胞增殖抑制作用(P<0.05)。与空白组比较,白花丹素组(1.5、3、6μmol·L^(-1)),T24细胞ROS、MDA及脂质过氧化物的含量明显升高,GSH水平明显降低,铁死亡相关蛋白SLC7A11、GPX4以及Nrf-2/Keap1明显降低(P<0.05)。结论:白花丹素能诱导细胞铁死亡,其机制与Nrf-2/Keap1信号通路有关。
