芹菜素对丙烯腈致大鼠肝内质网应激的影响

Effects of apigenin on endoplasmic reticulum stress induced by acrylonitrile in rats' hepatic tissue

目的探讨芹菜素(apigenin,AP)对丙烯腈(acrylonitrile,ACN)致大鼠肝内质网应激(ERS)信号通路的影响。方法 40只雄性SD大鼠随机分为对照组(玉米油)、ACN组(50 mg/kg·bw ACN)、低AP干预组(234 mg/kg·bw AP+50 mg/kg·bw ACN)和高AP干预组(468 mg/kg·bw AP+50 mg/kg·bw ACN),灌胃量均为5 ml/kg·bw,1次/d,每周6 d,共13周。末次染毒结束后,次日处死大鼠。分光光度法测定肝组织中丙二醛(MDA)、谷胱甘肽(GSH)含量和谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)活力;RT-PCR检测ERS相关基因GRP78、CHOP和Caspase-12 mRNA表达水平;Western blot检测ERS相关蛋白GRP78、CHOP和Caspase-12表达水平。结果 ACN组大鼠肝SOD活力、GSH-Px活力和GSH含量均低于对照组,MDA含量高于对照组,差异有统计学意义(P<0.05);低、高AP干预组大鼠肝SOD活力、GSH含量均高于ACN组,GSH-Px活力低于ACN组,差异有统计学意义(P<0.05)。ACN组大鼠肝组织GRP78、CHOP和Caspase-12 mRNA及蛋白表达水平均高于对照组(P<0.05)。低AP干预组大鼠肝组织GRP78、CHOP和Caspase-12 mRNA相对表达水平均低于ACN组(P<0.05),差异有统计学意义。高AP干预组大鼠肝组织GRP78和CHOP mRNA相对表达水平均低于ACN组(P<0.05),GRP78、Caspase-12蛋白表达水平均低于ACN组,差异有统计学意义(P<0.05)。结论 ACN可致大鼠肝氧化损伤,激活ERS信号通路,AP可减轻ACN引起的ERS。

Objective To study the effects of AP on endoplasmic reticulum stress(ERS） induced by acrylonitrile(ACN） in rats＇ liver. Methods Forty male SD rats were randomly divided into one control group(corn oil） and three experimental groups,50 mg/kg·bw ACN(ACN group）,234 mg/kg·bw AP ＋ 50 mg/kg·bw ACN(low dose AP intervention group）,468 mg/kg·bw AP ＋ 50 mg/kg·bw ACN(high dose AP intervention group）,respectively,rats were given by gavage once a day for 13 weeks with 5 ml/kg·bw liquid. At the end of the experiment,rats were executed and the contents of MDA and GSH,the activities of GSH-Px and SOD in hepatic tissue were detected by spectrophotometry. The mRNA expression levels of GRP78,CHOP and Caspase-12 were detected by RT-PCR. The protein level of GRP78,CHOP and Caspase-12 were detected by Western blot. Results The activities of SOD and GSH-Px were significantly decreased in ACN group than control group as well as the content of GSH. The content of MDA was signiticantly increased than control group(P〈 0. 05）. In low dose AP intervention group and high dose AP intervention group,the activities of SOD and the content of GSH were significantly increased,and the activity of GSH-Px was decreased compared with ACN group(P 〈0. 05）. In ACN group,the expression levels of GRP78,CHOP and Caspase-12 mRNA and protein were higher than control group. In low dose AP intervention group,the expressions of GRP78,CHOP and Caspase-12 mRNA were lower than ACN group,and the expression level of caspase-12 mRNA was decreased in low dose AP intervention group compared with ACN group(P 〈0. 05）. In high dose AP intervention group,the expression levels of GRP78 and CHOP mRNA were higher than control group,and the protein levels of GRP78,Caspase-12 were significantly decreased(P 〈0. 05）. Conclusion ACN-induced oxidative damage could activate ERS signaling pathway in rat＇s hepatic tissue,which can be alleviated by antioxidant AP.