文献类型：期刊文献

中文题名：芍药苷对乳腺炎小鼠乳腺组织氧化应激损伤的信号通路研究

英文题名：Research on the signaling pathway of paeoniflorin in oxidative stress damage of mammary gland tissue in mastitis mice

作者：刘馨鸿[1];郭超[1];宋冰[2,3];白敏[2,3];汪湛东[2,3];赵泓彰[2,3];王琼[2,3];张延英[2,3];赵兴绪[1]

第一作者：刘馨鸿

机构：[1]甘肃农业大学动物医学院,甘肃兰州730070;[2]甘肃中医药大学,甘肃兰州730000;[3]甘肃省实验动物行业技术中心,甘肃兰州730000

第一机构：甘肃农业大学动物医学院,甘肃兰州730070

年份：2024

卷号：46

期号：8

起止页码：847

中文期刊名：中国预防兽医学报

外文期刊名：Chinese Journal of Preventive Veterinary Medicine

收录：CSTPCD;;北大核心:【北大核心2023】；CSCD:【CSCD_E2023_2024】；

基金：甘肃省教育科技创新项目(2023A-080);国家自然科学基金联合基金重点项目(U21A20262)。

语种：中文

中文关键词：乳腺炎;芍药苷;Nrf2信号通路;氧化应激

外文关键词：mastitis;peony glucoside;Nrf2 pathway;oxidative damage

摘要：为探究芍药苷通过何种通路对乳腺炎小鼠氧化应激损伤的保护作用及作用机制,本研究选取产后6 d的BALB/c雌鼠,随机分为阴性对照组、模型组、芍药苷低剂量组(10 mg/kg)、芍药苷中剂量组(20 mg/kg)、芍药苷高剂量组(40 mg/kg)和地塞米松阳性对照组(5 mg/kg),每组10只,除阴性对照组外,其余小鼠均采用乳导管注入脂多糖(LPS)的方法建立乳腺炎模型,分别于造模前14 h、2 h以及造模后6 h对小鼠给药干预。造模结束24 h后剖检各组小鼠,观察乳腺组织形态变化,制备各组小鼠乳腺组织病理切片观察其病理变化。结果显示,与阴性对照组相比,模型组小鼠乳区红肿并伴有出血点,乳腺组织病变观察发现乳腺腺泡细胞内炎性细胞浸润明显,腺泡形态改变,表明乳腺炎小鼠模型可用于后续实验。取各组小鼠乳腺组织,采用荧光探针法测定小鼠乳腺组织中活性氧自由基(ROS)含量,采用免疫荧光组织化学技术测定小鼠乳腺组织中4-羟基壬烯酸(4-HNE)蛋白的表达水平,采用生化试剂盒检测小鼠乳腺组织中超氧化物歧化酶(SOD)、丙二醛(MDA)及谷胱甘肽过氧化物酶(GSH-Px)的含量,采用荧光定量PCR(RT-qPCR)和western blot分别检测小鼠乳腺组织中Nrf2信号通路关键分子核因子-红细胞2相关因子2(Nrf 2)、血红素氧合酶1(HO-1)、谷氨酸-半胱氨酸连接酶(GCLC)、醌氧化还原酶(NQO1)mRNA的转录及蛋白表达水平。结果显示,与阴性对照组相比,模型组小鼠乳腺组织中ROS、MDA含量及4-HNE蛋白表达水平均极显著升高(P<0.01),GSH-Px和SOD酶活性及含量均极显著降低(P<0.01),Nrf2、HO-1、GCLC和NQO1 mRNA的转录水平均显著或极显著上调(P<0.05或P<0.01),其蛋白表达水平与mRNA转录水平趋势基本一致;与模型组相比,各给药组小鼠乳腺组织病理损伤明显减轻,小鼠乳腺组织中ROS、MDA含量以及4-HNE蛋白表达水平显著或极显著降低(P<0.05或P<0.01),GSH-Px、SOD含量极显著或显著升高(P<0.01或P<0.05);Nrf2、HO-1、GCLC和NQO1mRNA的转录水平均显著或极显著上调(P<0.05或P<0.01),其蛋白表达水平与mRNA转录水平趋势基本一致。综上所述,本研究证实芍药苷能够治疗小鼠乳腺炎及小鼠乳腺组织氧化损伤,且高剂量芍药苷的治疗作用最佳,其具体作用机制与芍药苷显著激活宿主细胞中Nrf2信号通路拮抗氧化应激损伤有关,该结果为芍药苷在动物乳腺炎预防和治疗中的应用提供了科学依据。
To explore the protective effect and mechanism of paeoniflorin on oxidative stress-induced damage in mice with mastitis,female BALB/c mice 6 days postpartum were randomly assigned into 6 groups:a negative control group,a model group,and four treatment groups receiving varying doses of paeoniflorin(10mg/kg,20mg/kg,and 40mg/kg,respectively),as well as a positive control group(5mg/kg),with 8 mice in each group.Except for the negative control group,mastitis was induced in the remaining mice by intraductal injection of lipopolysaccharides(LPS).The mice were subjected to drug intervention 14 hours prior to induction,2 hours post-induction,and 6 hours after induction.24 hours following the induction of mastitis,the mice in each group were euthanized for examination of morphological alterations in the mammary gland tissue.Pathological sections of the mammary gland tissue in mice from each group were prepared to observe pathological changes.The levels of reactive oxygen species(ROS)in the mammary gland tissue of the mice were determined using a fluorescent probe method.The protein expression levels of 4-hydroxynonenal(4-HNE)in the mammary gland tissue of the mice were assessed via immunofluorescence histochemistry.The levels of superoxide dismutase(SOD),malondialdehyde(MDA),and glutathione peroxidase(GSH-Px)in the mammary gland tissue of the mice were quantified using biochemical assay kits.Real-time fluorescence quantitative PCR(RT-qPCR)and western blot were employed to detect the mRNA transcription and protein expression levels of key molecules in the nuclear factor erythroid2-related factor 2(Nrf2)pathway,including Nrf2,heme oxygenase-1(HO-1),glutamate-cysteine ligase(GClC),and NADH quinone oxidoreductase 1(NQO1)in the mammary gland tissue of the mice.The results showed that compared to the negative control group,the model group mice exhibited redness and bleeding points in the mammary gland area,and the pathological structure of mammary gland tissues revealed significant infiltration of inflammatory cells in the mammary acinar cells with morphological changes.Compared to the negative control group,the model group mice showed a marked elevation in ROS and MDA levels,along with increased 4-HNE protein expression within mammary gland tissues(P<0.01 or P<0.05).Conversely,the activity of GSH-Px and SOD enzymes were significantly decreased(P<0.01 or P<0.05).The transcription levels of Nrf2,HO-1,GCLC,and NQO1 mRNA were significantly upregulated(P<0.01 or P<0.05),with their protein expression levels showing a similar trend to mRNA transcription levels.In comparison to the model group,the treatment groups demonstrated a substantial reduction in pathological damage to mammary gland tissues.The protein expression levels of ROS,MDA,and 4-HNE in mammary gland tissues were significantly decreased(P<0.01),while GSH-Px and SOD were increased(P<0.01).The transcription levels of Nrf2,HO-1,GCLC,and NQO1 were all significantly upregulated(P<0.01 or P<0.05),with protein expression patterns paralleling those of mRNA transcription.The above results indicate that paeoniflorin effectively mitigate oxidative damage in the mammary tissue of mice with mastitis,with the high-dose group showing particularly optimal therapeutic effect.The specific mechanism appears to involve the significant activation of the Nrf2 pathway by paeoniflorin,which counteracts oxidative stress damage.