红芪多糖对ob/ob小鼠肝蛋白激酶R样内质网激酶-转录因子NF-E2相关因子2信号通路的影响  

Effects of Hedysarum polybotrys sacchcaide on protein kinase R-like endoplasmic reticulum kinase-transcription factor NF-E2-related factor 2 signaling pathway in liver of ob/ob mice

作　　者：张磊 金智生 万生芳 李亚玲 田一虹 杨秀娟 ZHANG Lei;JIN Zhi-sheng;WAN Sheng-fang;LI Ya-ling;TIAN Yi-hong;YANG Xiu-juan(School of Basic Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,Gansu Province,China;Key Laboratory of Dunhuang Medicine,Ministry of Education,Gansu University of Chinese Medicine,Lanzhou 730000,Gansu Province,China;Clinical College of Traditional Chinese Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,Gansu Province,China;School of Public Health,Gansu University of Chinese Medicine,Lanzhou 730000,Gansu Province,China;School of Pharmacy,Gansu University of Chinese Medicine,Lanzhou 730000,Gansu Province,China)

机构地区：[1]甘肃中医药大学基础医学院,甘肃兰州730000 [2]甘肃中医药大学敦煌医学与转化教育部重点实验室,甘肃兰州730000 [3]甘肃中医药大学中医临床学院,甘肃兰州730000 [4]甘肃中医药大学公共卫生学院,甘肃兰州730000 [5]甘肃中医药大学药学院,甘肃兰州730000

出　　处：《中国临床药理学杂志》2022年第17期2034-2038,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金地区基金资助项目(81660777);敦煌医学与转化教育部重点实验室开放课题基金资助项目(DHYX20-04);2021年度甘肃省科技计划(自筹经费)基金资助项目(21JR1RA274)。

摘　　要：目的 研究红芪多糖(HPS)对ob/ob小鼠肝组织葡萄糖调节蛋白78 (GRP78)、蛋白激酶R样内质网激酶-转录因子NF-E2相关因子2 (PERK-Nrf2)信号通路的影响。方法 将6周龄雄性C57BL/6 ob/ob小鼠随机分为5组:模型组(蒸馏水5 mL·kg^(-1)·d^(-1))、对照组(硫辛酸30 mg·kg^(-1)·d^(-1))和高、中、低剂量实验组(红芪多糖200、100、50 mg·kg^(-1)·d^(-1)),每组10只;另选非转基因C57BL/6正常雄性小鼠10只正常组(蒸馏水5 mL·kg^(-1)·d^(-1)),连续灌胃8周。用逆转录聚合酶链反应(RT-PCR)、蛋白质印迹(Western blot)法检测肝组织GRP78、Nrf2 mRNA和蛋白的表达,以蛋白质印迹法检测PERK、p-Nrf2和p-PERK蛋白的表达,计算Nrf2、PERK磷酸化水平。结果 正常组、模型组、对照组和高、中、低剂量实验组肝组织中GRP78 mRNA表达量分别为1.00±0.00,1.40±0.15,0.99±0.03,1.02±0.08,1.21±0.05,1.31±0.17;肝组织中Nrf2 mRNA表达量分别为1.00±0.00,0.18±0.02,1.01±0.03,1.00±0.13,0.90±0.10,0.38±0.05,Nrf2蛋白磷酸化(p-Nrf2/Nrf2)水平分别为0.91±0.01,1.28±0.08,1.10±0.13,0.91±0.08,1.09±0.02,1.24±0.08;PERK蛋白磷酸化(p-PERK/PERK)水平分别为0.88±0.06,0.15±0.01,0.65±0.08,0.68±0.04,0.46±0.06,0.26±0.01,上述指标:与正常组相比,模型组GRP78 mRNA和Nrf2蛋白磷酸化水平均升高,Nrf2 mRNA和PERK蛋白磷酸化水平均下降(均P<0.01),与模型组相比,对照组及实验各剂量组GRP78 mRNA和Nrf2蛋白磷酸化水平均降低,Nrf2 mRNA和PERK蛋白磷酸化水平均升高(均P<0.01)。结论 红芪多糖可能通过调控GRP78表达,并通过PERK-Nrf2信号通路提高肝抗氧化能力,减轻内质网应激,从而保护ob/ob小鼠肝组织。Objective To study the effect of Hedysarum polybotrys sacchcaide(HPS) on glucose regulated protein 78(GRP78) and protein kinase R-like endoplasmic reticulum kinase-transcription factor NF-E2-related factor2(PERK-Nrf2) signal pathway in the liver of ob/ob mice.Methods Male C57 BL/6 ob/ob mice aged 6 weeks were randomly divided into 5 groups:model group(distilled water 5 mL·kg^(-1)·d^(-1)),positive control group(lipoic acid 30 mg·kg^(-1)·d^(-1)) and high,medium and low dose experimental groups( HPS 200,100,50 mg · kg^(-1)·d^(-1)) with 10 rats in each group;10 non-transgenic C57BL/6 normal male mice were selected as normal group( distilled water 5 m L·kg^(-1)·d^(-1)).Each group was given intragastric intervention continuously for 8 weeks.The expressions of GRP78,Nrf2m RNA and protein in liver were detected by reserve transtription polymerse chain reaction( RT-PCR) and Western blot,the protein expressions of PERK,p-Nrf2 and p-PERK were detected by Western blot,and the phosphorylation levels of Nrf2 and PERK were calculated.Results The expression of GRP78 m RNA in normal group,model group,positive control group and high,medium and low dose experimental groups were 1.00 ± 0.00,1.40 ± 0.15,0.99 ± 0.03,1.02 ± 0.08,1.21 ± 0.05,1.31 ± 0.17,respectively;the expression of Nrf2 m RNA were 1.00 ± 0.00,0.18 ± 0.02,1.01 ± 0.03,1.00 ± 0.13,0.90 ± 0.10,0.38 ± 0.05,respectively;the phosphorylation levels of Nrf2( p-Nrf2/Nrf2) were 0.91 ± 0.01,1.28 ± 0.08,1.10 ± 0.13,0.91 ± 0.08,1.09 ± 0.02,1.24 ± 0.08;the phosphorylation levels of PERK( p-PERK/PERK) were 0.88 ± 0.06,0.15 ± 0.01,0.65 ± 0.08,0.68 ± 0.04,0.46 ± 0.06,0.26 ± 0.01.The above indexes:compared with the normal group,the expression of GRP78 m RNA and p-Nrf2/Nrf2 in model group were significantly increased( P < 0.01),the expression of Nrf2 m RNA and p-PERK/PERK in the model group were significantly decreased( P < 0.01),compared with the model group,the expression of GRP78 m RNA and p-Nrf2/Nrf2 in the positive control group and experimental group

关 键 词：红芪多糖 非酒精性脂肪性肝病 内质网应激 葡萄糖调节蛋白78 蛋白激酶R样内质网激酶 转录因子NF-E2相关因子2 

分 类 号：R28[医药卫生—中药学]