原花青素对氯氰菊酯致小鼠小脑组织氧化损伤及核转录因子NF-E2相关因子2、血红素氧合酶-l表达的影响
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摘要
目的研究原花青素(proanthocyanidin,PC)对氯氰菊酯(cypermethrin,CYP)所致小鼠小脑组织氧化损伤及核转录因子NF-E2相关因子2(Nrf2)、血红素氧合酶-l(HO-1)表达的影响。方法将50只雄性昆明小鼠随机分为5组:双蒸水对照组,染毒组(以CYP 10 mg/kg进行灌胃),PC保护组Ⅰ、Ⅱ、Ⅲ(分别以50、100和200mg/kg PC预先给小鼠灌胃,再以10 mg/kg CYP染毒)。连续3周经口灌胃,之后颈椎脱臼法处死小鼠,检测小脑组织谷胱甘肽过氧化物酶(GSH-Px)、总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)活性和丙二醛(MDA)含量;免疫组化分析Nrf2在小脑组织中的表达;RT-PCR检测Nrf2及HO-1 mRNA的表达。结果染毒组小鼠小脑组织MDA含量升高,GSH-Px、T-SOD、CAT活性降低;Nrf2胞核阳性细胞比例增加;Nrf2及HO-1 mRNA表达增加,与对照组比较差异具有统计学意义(P<0.05)。与染毒组相比,PC保护组MDA水平降低,T-SOD、GSH-Px、CAT活性增加;胞核Nrf2阳性细胞逐渐减少;Nrf2及HO-1 mRNA表达下降,与染毒组比较差异具有统计学意义(P<0.05)。结论 CYP暴露可诱导小鼠小脑组织氧化损伤,使Nrf2由胞质向胞核转位,同时诱导下游靶基因HO-1的表达;抗氧化剂PC能够使活化的Nrf2及HO-1mRNA表达回调至基础水平,从而拮抗CYP所致的氧化损伤。
Objective To investigate the protective effect of proanthocyanidin on oxidative stress and mRNA expression of Nrf2 and HO-1 of mice cerebellar tissue induced by cypermethrin. Methods A total of 50 male Kunming mice were randomly divided into five groups by gavage once daily for three consecutive weeks,the control group,CYP group( 10 mg / kg) and PC pretreated group( 50,100 and 200 mg / kg). The mice were sacrificed and activitie levels of GSH-Px,T-SOD,CAT and MDA were measured,and then used the immunohistochemistry to analyse the expression of Nrf2,semi-quantitative RTPCR detected mRNA expression of Nrf2 and HO-1. Results Compared to the controlgroup,the activity levels of GSH-Px,T-SOD,CAT decreased and MDA increased in CYP group. Nucleus positive cells was higher and mRNA expression of Nrf2 and HO-1 was higher than the control group( P < 0. 05). Compared to the CYP group,the level of GSHPx、T-SOD、CAT increased and MDA decreased in PC pretreated group,the count of nucleus-positive cells and mRNA expression of Nrf2 and HO-1 was lower than the CYP group( P < 0. 05). Conclusions Cypermethrin could increase oxidative stress in cerebellar tissue of mice,induces the nuclear translocation of Nrf2 from the cytoplasm to the nucleus,and upregulating downstream gene of HO-1 expression. Proanthocyanidin could adjust mRNA expression of Nrf2 and HO-1 and ameliorate cypermethrin-induced oxidative stress.
Objective To investigate the protective effect of proanthocyanidin on oxidative stress and mRNA expression of Nrf2 and HO-1 of mice cerebellar tissue induced by cypermethrin. Methods A total of 50 male Kunming mice were randomly divided into five groups by gavage once daily for three consecutive weeks,the control group,CYP group( 10 mg / kg) and PC pretreated group( 50,100 and 200 mg / kg). The mice were sacrificed and activitie levels of GSH-Px,T-SOD,CAT and MDA were measured,and then used the immunohistochemistry to analyse the expression of Nrf2,semi-quantitative RTPCR detected mRNA expression of Nrf2 and HO-1. Results Compared to the controlgroup,the activity levels of GSH-Px,T-SOD,CAT decreased and MDA increased in CYP group. Nucleus positive cells was higher and mRNA expression of Nrf2 and HO-1 was higher than the control group( P < 0. 05). Compared to the CYP group,the level of GSHPx、T-SOD、CAT increased and MDA decreased in PC pretreated group,the count of nucleus-positive cells and mRNA expression of Nrf2 and HO-1 was lower than the CYP group( P < 0. 05). Conclusions Cypermethrin could increase oxidative stress in cerebellar tissue of mice,induces the nuclear translocation of Nrf2 from the cytoplasm to the nucleus,and upregulating downstream gene of HO-1 expression. Proanthocyanidin could adjust mRNA expression of Nrf2 and HO-1 and ameliorate cypermethrin-induced oxidative stress.
引文
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[15]LI S G,DING Y S,NIU Q,et al.Grape seed proanthocyanidin extract alleviates arsenic induced oxidative reproductive toxicity in male mice[J].Biomed Environ Sci,2015,28(4):272-280.
[16]MANSOURI E,KHORSANDI L,ABDOLLAHZADE FARD A.Protective role of grape seed proanthocyanidin antioxidant properties on heart of streptozotocin-induced diabetic rats[J].Vet Res Forum,2015,6(2):119-124.
[2]NASUTI C,GABBIANELLI R,FALCIONI M L,et al.Dopaminergic system modulation,behavioral changes,and oxidative stress after neonatal administration of pyrethroids[J].Toxicology,2007,229(3):194-205.
[3]SINGH A K,TIWARI M N,PRAKASH O,et al.A current review of cypermethrin-induced neurotoxicity and nigrostriatal dopaminergic neurodegeneration[J].Curr Neuropharmacol,2012,10(1):64-71.
[4]CAO D,CHEN N,ZHU C,et al.β-cypermethrininduced acute neurotoxicity in the cerebral cortex of mice[J].Drug Chem Toxicol,2015,8(1):44-49.
[5]NGUYEN T,NIOI P,PICKETT C B.The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress[J].J Biol Chem,2009,284(20):13291-13295.
[6]RENE C,LOPEZ E,CLAUSTRES M,et al.NF-E2-related factor 2,a key inducer of antioxidant defenses,negatively regulates the CFTR transcription[J].Cell Mol Life Sci,2010,67(13):2297-2309.
[7]BAGCHI D,BAGCHI M,STOHS S J,et al.Free radicals and grape seed proanthocyanidin extract:importance in human health and disease prevention[J].Toxicology,2000,148(2-3):187-197.
[8]ZHANG Z,ZHENG L,ZHAO Z.Grape seed proanthocyanidins inhibit H2O2-induced osteoblastic MC3T3-E1 cell apoptosis via ameliorating H2O2-induced mitochondrial dysfunction[J].J Toxicol Sci,2014,39(5):803-813.
[9]HAYES J D,MCMAHON M.NRF2 and KEAP1mutations:permanent activation of an adaptive response in cancer[J].Trends Biochem Sci,2009,34(4):176-188.
[10]LIU Q,ZHANG H,SMEESTER L,et al.The NRF2-mediated oxidative stress response pathway is associated with tumor cell resistance to arsenic trioxide across the NCI-60 panel[J].BMC Med Genomics,2010,8(3):37-41.
[11]SHEN G,KONG A N.Nrf2 plays an important role in coordinated regulation of Phase II drug metabolism enzymes and Phase III drug transporters[J].Biopharm Drug Dispos,2009,30(7):345-355.
[12]CHEN S,ZHU Y,LIU Z,et al.Grape seed proanthocyanidin extract ameliorates diabetic bladder dysfunction via the activation of the Nrf2 pathway[J].PLo S One,2015,10(5):e0126457.
[13]WANG G,XIU P,LI F.Vitamin A supplementation alleviates extrahepatic cholestasis liver injury through Nrf2 activation[J].Oxid Med Cell Longev,2014:273-692.
[14]LIU Y,ZHANG L,LIANG J.Activation of the Nrf2defense pathway contributes to neuroprotective effects of phloretin on oxidative stress injury after cerebral ischemia/reperfusion in rat[J].J Neurol Sci,2015,351(1-2):88-92.
[15]LI S G,DING Y S,NIU Q,et al.Grape seed proanthocyanidin extract alleviates arsenic induced oxidative reproductive toxicity in male mice[J].Biomed Environ Sci,2015,28(4):272-280.
[16]MANSOURI E,KHORSANDI L,ABDOLLAHZADE FARD A.Protective role of grape seed proanthocyanidin antioxidant properties on heart of streptozotocin-induced diabetic rats[J].Vet Res Forum,2015,6(2):119-124.