Background:Epidemiological studies have confirmed that longer exposure to insecticides like cypermethrin(CYP)significantly increases the risk of male reproductive toxicity.Crocus sativus L.has been recognized due to i...Background:Epidemiological studies have confirmed that longer exposure to insecticides like cypermethrin(CYP)significantly increases the risk of male reproductive toxicity.Crocus sativus L.has been recognized due to its therapeutic properties,but its exact role and molecular mechanisms in treatment of reproductive dysfunction remain unclear.Methods:During this study,36 rats were randomly divided into six groups(n=6):control,CYP-induced(60 mg/kg),standard(leuprolide 3 mg/kg)and three treatment groups receiving aqueous,ethanolic,and oil extracts(50 mg/kg or 20 mL/kg)for post-toxicity induction.Results:The finding represented that exposure of CYP significantly increased oxidative stress,disrupted testicular architecture,and markedly reduced testosterone levels(P<0.05).Importantly,Crocus sativus L.treatment alleviated these changes by increasing the expression of Nrf2(nuclear factor erythroid 2-related factor 2),restoring the activity of antioxidant enzymes,and enhancing testicular histomorphology.Surprisingly,molecular docking established a high binding affinity of Crocus sativus L.phytoconstituents such as gallic acid,cinnamic acid and quercetin to the Nrf2-Keap1 complex.It is worth noting that,Crocus sativus L.exhibited a high level of protection against reproductive toxicity caused by CYP in male rats,which was mediated by the activation of Nrf2 pathway,reduction of oxidative damage,and favorable ADMET characteristics.Conclusion:Notably,this research provides a more valid,safe,and effective method of developing new drugs for reproductive disorders,however,further investigation is needed to support the research findings and implement it in clinical practice.展开更多
文摘Background:Epidemiological studies have confirmed that longer exposure to insecticides like cypermethrin(CYP)significantly increases the risk of male reproductive toxicity.Crocus sativus L.has been recognized due to its therapeutic properties,but its exact role and molecular mechanisms in treatment of reproductive dysfunction remain unclear.Methods:During this study,36 rats were randomly divided into six groups(n=6):control,CYP-induced(60 mg/kg),standard(leuprolide 3 mg/kg)and three treatment groups receiving aqueous,ethanolic,and oil extracts(50 mg/kg or 20 mL/kg)for post-toxicity induction.Results:The finding represented that exposure of CYP significantly increased oxidative stress,disrupted testicular architecture,and markedly reduced testosterone levels(P<0.05).Importantly,Crocus sativus L.treatment alleviated these changes by increasing the expression of Nrf2(nuclear factor erythroid 2-related factor 2),restoring the activity of antioxidant enzymes,and enhancing testicular histomorphology.Surprisingly,molecular docking established a high binding affinity of Crocus sativus L.phytoconstituents such as gallic acid,cinnamic acid and quercetin to the Nrf2-Keap1 complex.It is worth noting that,Crocus sativus L.exhibited a high level of protection against reproductive toxicity caused by CYP in male rats,which was mediated by the activation of Nrf2 pathway,reduction of oxidative damage,and favorable ADMET characteristics.Conclusion:Notably,this research provides a more valid,safe,and effective method of developing new drugs for reproductive disorders,however,further investigation is needed to support the research findings and implement it in clinical practice.
文摘目的探究枸杞多糖(Lycium barbarum polysaccharides,LBP)调控核因子E2相关因子2(nuclear factor E2 related factor 2,Nrf2)/谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)通路对缺氧/复氧(hypoxia/reoxygenation,H/R)诱导的心肌细胞铁死亡的影响。方法设置对照组、模型组及LBP低、中、高剂量(25、50、100 mg/L)组和LBP(100mg/L)+Nrf2抑制剂ML385(5μmol/L)组。构建H9c2细胞H/R模型,并给予相应药物处理。采用CCK-8检测细胞活力;采用TUNEL染色检测细胞凋亡;采用试剂盒检测细胞Fe^(2+)、谷胱甘肽(glutathione,GSH)、丙二醛(malondialdehyde,MDA)水平及乳酸脱氢酶(lactate dehydrogenase,LDH)、超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)活性;采用二氢乙锭(dihydroethidium,DHE)荧光探针检测细胞活性氧(reactive oxygen species,ROS)水平;采用透射电镜观察细胞线粒体形态;采用Western blotting检测转铁蛋白受体1(transferrin receptor 1,Tf R1)、二价金属离子转运蛋白1(divalent metal transporter 1,DMT1)、铁蛋白重链1(ferritin heavy chain 1,FTH1)、酰基辅酶A合成酶长链家族成员4(acyl-CoA synthetase long-chain family member 4,ACSL4)、Kelch样ECH相关蛋白1(Kelch like ECH associated protein1,Keap1)、Nrf2、血红素氧合酶-1(heme oxygenase-1,HO-1)、溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)、GPX4蛋白表达。结果与对照组比较,模型组细胞活力显著降低(P<0.001),Fe^(2+)、MDA、ROS水平和LDH活性显著升高(P<0.001),GSH水平和SOD、CAT活性显著降低(P<0.001),线粒体缩短、膜密度增高且嵴数量减少,TfR1、Nrf2、HO-1、SLC7A11、GPX4蛋白表达水平显著降低(P<0.001),DMT1、FTH1、ACSL4、Keap1蛋白表达水平显著升高(P<0.001)。与模型组比较,LBP组细胞活力显著升高(P<0.01),Fe^(2+)、MDA、ROS水平和LDH活性显著降低(P<0.05、0.01、0.001),GSH水平和SOD、CAT活性显著升高(P<0.05、0.01、0.001),线粒体形态较为规则,TfR1、Nrf2、HO-1、SLC7A11、GPX4蛋白表达水平显著升高(P<0.01、0.001),DMT1、FTH1、ACSL4、Keap1蛋白表达水平显著降低(P<0.05、0.01、0.001);而ML385能够抑制枸杞多糖对细胞铁死亡的改善作用(P<0.05、0.01、0.001)。结论枸杞多糖可能通过激活Nrf2/GPX4通路改善H/R诱导的心肌细胞铁死亡。
