芦丁对湖羊乳腺氧化应激相关指标及核因子E2相关因子2信号通路基因表达的影响

Effects of Rutin on Oxidative Stress-Related Indices and Nuclear Factor E2 Related Factor 2 Signaling Pathway Related Gene Expression in Mammary Gland of Hu Sheep

本试验旨在研究饲粮中添加芦丁对产后湖羊乳腺氧化应激相关指标及核因子E2相关因子2(NRF2,基因为NFE2L2)信号通路基因表达的影响。选取体重相近且分娩日期相近的湖羊30只,随机分为3组,每组10只。对照组饲喂基础饲粮,试验组分别饲喂在基础饲粮中添加50和100 mg/kg BW芦丁的饲粮。预试期7 d,正试期56 d。结果表明:1)与对照组相比,饲粮中添加50 mg/kg BW芦丁能够减少湖羊乳腺腺泡内炎性细胞数量,100 mg/kg芦丁组湖羊乳腺腺泡内未见炎性细胞。2)与对照组相比,饲粮中添加100 mg/kg BW芦丁显著或极显著提高乳腺组织中过氧化氢酶(CAT)(P<0.01)和谷胱甘肽过氧化物酶(GSH-Px)(P<0.05)活性以及总抗氧化能力(T-AOC)(P<0.05),饲粮中添加50和100 mg/kg BW芦丁极显著降低过乳腺组织中过氧化氢(H_(2)O_(2))含量(P<0.01)。3)与对照组相比,饲粮中添加100 mg/kg BW芦丁极显著提高乳腺组织细胞内NRF2的蛋白表达水平(P<0.01),显著提高乳腺组织中NFE2L2 mRNA相对表达量(P<0.05),显著降低乳腺组织中Kelch样ECH关联蛋白1(KEAP1)mRNA相对表达量(P<0.05)。4)与对照组相比,饲粮中添加50 mg/kg BW芦丁显著提高乳腺组织中谷氨酸-半胱氨酸连接酶修饰亚基(GCLM)mRNA相对表达量(P<0.05);饲粮中添加100 mg/kg BW芦丁显著或极显著提高乳腺组织中GCLM(P<0.01)、GSH-Px(P<0.05)、硫氧还蛋白(TXN)(P<0.05)、铁蛋白重链1(FTH1)(P<0.01)、转醛醇酶1(TALDO1)(P<0.05)、苹果酸酶1(ME1)(P<0.05)和血红素氧合酶1(HMOX1)mRNA相对表达量,显著提高乳腺组织中B淋巴细胞瘤2(BCL2)mRNA相对表达量(P<0.05)。由此可见,饲粮中添加50和100 mg/kg BW芦丁能够减少湖羊乳腺腺泡内炎性细胞数量,并发挥抗氧化应激作用,通过调节NRF2信号通路的基因表达水平,改善产后湖羊乳腺的健康状态。

The purpose of the present study was to investigate the effects of rutin on oxidative stress-related indices and nuclear factor E2 related factor 2(NRF2,gene named NFE2L2)signaling pathway gene expression in mammary gland of postpartum Hu sheep.Thirty Hu sheep with similar body weight and similar birth date were randomly divided into 3 groups with 10 sheep in each group.Sheep in the control group were fed a basal diet,and those in the experimental groups were fed the basal diet supplemented with 50 and 100 mg/kg BW rutin,respectively.The pre-trial period lasted for 7 days and the experimental period lasted for 56 days.The results showed as follows:1)compared with the control group,dietary 50 mg/kg BW rutin could reduce the number of inflammatory cells in mammary gland acinus of Hu sheep,and no inflammatory cells were found in mammary gland acinus of Hu sheep in 100 mg/kg rutin group.2)Compared with the control group,dietary 100 mg/kg BW rutin significantly or extremely significantly increased the activities of catalase(CAT)(P<0.01)and glutathione peroxidase(GSH-Px)(P<0.05)and total antioxidant capacity(T-AOC)(P<0.05)in mammary gland,dietary 50 and 100 mg/kg BW rutin extremely significantly decreased the hydrogen peroxide(H_(2)O_(2))content in mammary gland(P<0.01).3)Compared with the control group,dietary 100 mg/kg BW rutin extremely significantly increased the NRF2 protein expression level in mammary gland cells(P<0.01),significantly increased the NFE2L2 mRNA relative expression level in mammary gland(P<0.05),and significantly decreased the Kelch-like ECH associated protein 1(KEAP1)mRNA relative expression level of in mammary gland(P<0.05).4)Compared with the control group,dietary 50 mg/kg BW rutin significantly increased the glutamate-cysteine ligase modified subunit(GCLM)mRNA relative expression level in mammary gland(P<0.05);dietary 100 mg/kg BW rutin significantly or extremely significantly increased the mRNA relative expression levels of GCLM(P<0.01),GSH-Px(P<0.05),thioredoxin(TXN)(P<0.05),ferritin heavy chain 1(FTH1)(P<0.01),transaldolase 1(TALDOL1)(P<0.05),malic enzyme 1(ME1)(P<0.05)and heme oxygenase 1(HMOX1)in mammary gland,and significantly increased the B lymphocytoma-2(BCL2)mRNA relative expression level in mammary gland(P<0.05).In conclusion,dietary 50 and 100 mg/kg BW rutin can reduce the number of inflammatory cells in mammary gland acinus of Hu sheep,play an antioxidant stress role,and improve the health status of postpartum mammary gland of Hu sheep by regulating the gene expression levels of NRF2 signaling pathway.