文献类型：期刊文献

中文题名：基于Nrf2/HO-1/GPX4信号通路探讨真武汤改善脾肾阳虚型糖尿病肾病小鼠的作用机制

英文题名：Zhenwutang Ameliorates Diabetic Kidney Disease in Mice with Spleen-kidney Yang Deficiency via Nrf2/HO-1/GPX4 Signaling Pathway

作者：赵哲[1,2];陈光顺[1];白敏[1,2];靳玉秋[1];田萌媛[1];宋冰[1,2];刘臻华[1];张延英[1,2]

第一作者：赵哲

机构：[1]甘肃中医药大学,兰州730000;[2]甘肃省实验动物行业技术中心,兰州730000

第一机构：甘肃中医药大学

年份：2024

卷号：30

期号：15

起止页码：29

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;Scopus;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；

基金：国家自然科学基金地区基金项目(82160865);甘肃省优秀研究生“创新之星”基金项目(2022CXZX-769);甘肃省自然科学基金项目(23JRRA1227)。

语种：中文

中文关键词：真武汤;脾肾阳虚;糖尿病肾病;核因子E2相关因子2(Nrf2)/血红素氧合酶-1(HO-1)/谷胱甘肽过氧化物酶4(GPX4)信号通路;氧化损伤

外文关键词：Zhenwutang;spleen-kidney Yang deficiency;diabetic kidney disease;nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase 1(HO-1)/glutathione peroxidase 4(GPX4)signaling pathway;oxidative damage

摘要：目的:基于核因子E2相关因子2(Nrf2)/血红素氧合酶-1(HO-1)/谷胱甘肽过氧化物酶4(GPX4)信号通路探讨真武汤对脾肾阳虚型糖尿病肾病(DKD)小鼠肾脏氧化损伤的干预作用及机制。方法:将25只7周龄SPF级雄性db/m小鼠及95只7周龄SPF级雄性db/db小鼠适应性喂养1周。db/m小鼠作为空白组,其余小鼠进行运用灌服生大黄溶液和氢化可的松制备脾肾阳虚型DKD模型,成模后随机分为模型组、厄贝沙坦组(25 mg·kg^(-1))、真武汤高、中、低剂量(33.8、16.9、8.45 g·kg^(-1))组,每组15只,连续灌胃8周。观察小鼠生存状态和计算中医证候评分并测定脾肾阳虚相关、空腹血糖(FBG)及肾功能相关指标;苏木素-伊红(HE)染色观察各组肾脏组织病理学改变;生化试剂盒法测定肾组织中氧化应激相关指标;实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测各组小鼠肾组织中Nrf2、HO-1、谷氨酸半胱氨酸连接酶催化亚基(GCLC)、GPX4 m RNA和蛋白表达水平。结果:与空白组比较,模型组小鼠中医证候积分明显升高(P<0.05),雌二醇(E2)含量明显升高(P<0.05),睾酮(T)、三碘甲状腺原氨酸(T3)、四碘甲状腺原氨酸(T4)含量明显降低(P<0.05);FBG明显升高(P<0.05),肾功能明显下降(P<0.05);肾脏病理显示肾小球肥大,肾小球系膜和基底增厚明显,肾组织中过氧化氢酶(CAT)、总抗氧化能力(T-AOC)、还原型谷胱甘肽(GSH)水平均明显降低(P<0.05),丙二醛(MDA)的含量明显升高(P<0.05),肾组织中Nrf2、HO-1、GCLC、GPX4 m RNA及蛋白表达水平明显下降(P<0.05);与模型组比较,真武汤高、中剂量组中医证候积分明显降低、E2含量明显降低(P<0.05),T、T3、T4含量明显升高(P<0.05);肾功能明显改善(P<0.05),肾脏病理明显改善,真武汤高、中剂量组小鼠肾组织中CAT、T-AOC、GSH水平均明显升高(P<0.05),MDA的含量明显下降(P<0.05),各给药组小鼠肾组织中Nrf2、HO-1、GCLC、GPX4 m RNA及蛋白表达水平明显升高(P<0.05)。结论:真武汤改善脾肾阳虚型db/db小鼠一般状态、肾功能,降低氧化损伤和减轻肾脏病理改变,其作用机制可能与调节Nrf2/HO-1/GPX4通路有关。
Objective:To investigate the effect and mechanism of Zhenwutang on renal oxidative damage in the mouse model of diabetic kidney disease with the syndrome of spleen-kidney Yang deficiency via the nuclear factor erythroid 2-related factor-2(Nrf2)/heme oxygenase-1(HO-1)/glutathione peroxidase 4(GPX4)signaling pathway.Method:Twenty-five 7-week-old SPF-grade male db/m mice and 957-week-old SPF-grade male db/db mice were adaptively fed for a week.A blank group was set with the db/m mice without treatment,and the other mice were administrated with Rhei Radix et Rhizoma decoction and hydrocortisone for the modeling of diabetic kidney disease with the syndrome of spleen-kidney Yang deficiency.The modeled mice were randomized into the model,irbesartan(25 mg·kg^(-1)),and high-,medium-,low-dose(33.8,16.9,8.45 g·kg^(-1))Zhenwutang groups(n=15)and administrated with corresponding drugs for 8 weeks.The survival status of mice was observed,and the traditional Chinese medicine(TCM)syndrome score was recorded.The indicators related to spleen-kidney Yang deficiency,fasting blood glucose(FBG),and renal function indicators were determined.Hematoxylin-eosin staining was employed to observe the histopathological changes of the renal tissue in each group.Biochemical kits were used to determine the oxidative stress-related indicators in the renal tissue.Realtime polymerase chain reaction and Western blotting were employed to determine the mRNA and protein levels,respectively,of Nrf2,HO-1,glutamate-cysteine ligase catalytic subunit(GCLC),and GPX4 in the renal tissue of mice in each group.Result:Compared with the blank group,the modeling increased the TCM syndrome score(P<0.05),elevated the estradiol(E2)and FBG levels(P<0.05),lowered the testosterone(T),triiodothyronine(T3),and tetraiodothyronine(T4)levels(P<0.05),and weakened the renal function(P<0.05).In addition,the modeling led to glomerular hypertrophy and glomerular mesangial and basal thickening,decreased the catalase(CAT)activity,total antioxidant capacity(T-AOC),and glutathione(GSH)content(P<0.05),increased the malondialdehyde(MDA)content(P<0.05),and down-regulated the mRNA and protein levels of Nrf2,HO-1,GCLC,and GPX4 in the renal tissue(P<0.05).Compared with the model group,high and medium doses of Zhenwutang decreased the TCM syndrome score and E2 content(P<0.05),increased the T,T3,and T4 content(P<0.05),improved the renal function(P<0.05),alleviated the pathological changes in the renal tissue,increased CAT,T-AOC,and GSH(P<0.05),reduced MDA(P<0.05),and up-regulated the mRNA and protein levels of Nrf2,HO-1,GCLC,and GPX4 in the renal tissue(P<0.05).Conclusion:Zhenwutang can improve the general state and renal function and reduce the oxidative damage and pathological changes in the renal tissue of db/db mice with spleen-kidney Yang deficiency by regulating the Nrf2/HO-1/GPX4 signaling pathway.