穿山龙总皂苷经Nrf2/ARE通路抑制慢性肾脏病大鼠血管钙化的作用  

Inhibition effect of total saponin from rhizoma dioscorea nipponica on vascular calcification in rats with chronic kidney disease via Nrf2/ARE pathway

作　　者：马娟 郭银雪 谢恂 张晶晶 黄宁川 MA Juan;GUO Yin-Xue;XIE Xun(Department of Nephrology,the First Affiliated Hospital of Guizhou University of traditional Chinese medicine,Guiyang 550000,Guizhou,China)

机构地区：[1]贵州中医药大学第一附属医院肾内科,贵州贵阳550000

出　　处：《中国老年学杂志》2022年第12期3011-3016,共6页Chinese Journal of Gerontology

基　　金：贵州省科技计划项目(黔科合LH字[2017]7051号)。

摘　　要：目的 探讨穿山龙总皂苷(TSRDN)经核因子E2相关因子(Nrf)2/抗氧化反应元件(ARE)通路抑制慢性肾脏病(CKD)大鼠血管钙化的作用。方法 50只SPF级成年雄性SD大鼠,随机分为对照组、模型组、TSRDN低剂量组、TSRDN高剂量组和骨化三醇组,每组10只,对照组用普通饲料喂养,其余各组用腺嘌呤(250 mg/kg, 1次/d)灌胃和含1.8%高磷饲料喂养,连续4 w。第5~8周腺嘌呤改为隔日灌胃,同时TSRDN低剂量组和TSRDN高剂量组分别灌胃给予TSRDN(80 mg/kg、160 mg/kg),骨化三醇组灌胃给予骨化三醇(0.045μg/kg),灌胃体积3 ml,对照组和模型组灌胃给予等体积生理盐水。每天给药1次,连续给药8 w后股动脉取血用自动生化分析仪检测钙(Ca)、磷(P)、血清肌酐(Scr)和尿素氮(BUN)水平,然后取肾脏和主动脉组织进行苏木素-伊红(HE)染色观察组织学变化,采用实时qRT-PCR检测主动脉中Nrf2、血红素加氧酶(HO)-1、磷酸酰胺腺嘌呤二核苷酸醌氧化还原酶(NQO)-1、γ-谷氨酰半胱氨酸合成酶(GCS)、核心结合蛋白因子(RUNX)2和骨形态发生蛋白(BMP)2 mRNA水平。结果 对照组肾脏和主动脉血管结构正常;模型组肾脏和主动脉结构破坏严重,见大量黑色腺嘌呤代谢物结晶沉积,在动脉中膜处见连续性线性分布的明显钙化结节;与模型组比较,各TSRDN剂量组和骨化三醇组大鼠肾脏病变明显减轻,腺嘌呤结晶减少,血管结构破坏减轻,钙化结节减少,TSRDN高剂量组和骨化三醇组较为明显。与对照组比较,模型组、各TSRDN剂量组和骨化三醇组Ca、Nrf2、HO-1、NQO-1和γ-GCS mRNA水平降低,P、Scr、BUN、RUNX2和BMP2 mRNA水平升高(P<0.05);与模型组相比,各TSRDN剂量组和骨化三醇组Ca、Nrf2、HO-1、NQO-1和γ-GCS mRNA水平升高,P、Scr、BUN、RUNX2和BMP2 mRNA水平降低,且呈剂量依赖性(P<0.05);TSRDN高剂量组和骨化三醇组作用相似(P>0.05)。结论 TSRDN能改善CKD大鼠肾功能和钙磷代谢,抑制血管钙化,其Objective To investigate the inhibition effect of total saponin from rhizoma dioscorea nipponica(TSRDN)on vascular calcification in rats with chronic kidney disease via nuclear factor erythroid E2-related factor(Nrf2)/antioxidant response element(ARE)pathway.Methods 50 SPF grade adult male SD rats were randomly divided into control group,model group,TSRDN low dose group,TSRDN high dose group and calcitriol group,10 rats in each group.The control group was fed with ordinary feed,and the remaining groups were fed with adenine(250 mg/kg,once/day)by gavage and fed with 1.8%high phosphorus feed for 4 weeks.At weeks 5~8,adenine was changed to gavage every other day.At the same time,TSRDN(80 mg/kg,160 mg/kg)was administered orally to TSRDN low dose group and TSRDN high dose group by gavage.In the calcitriol group,calcitriol(0.045μg/kg)was given by gavage,gavage volume 3 ml,the control group and the model group were orally administered with an equal volume of normal saline,once a day,continuous administration for 8 weeks.Blood samples were taken from the femoral artery,and the levels of calcium(Ca),phosphorus(P),serum creatinine(Scr),and urea nitrogen(BUN)were measured with an automatic biochemical analyzer,and then kidneys and aortic tissue were taken for hematoxylin-eosin(HE)staining to observe histological changes,and real-time qRT-PCR was used to detect Nrf2,heme oxygenase(HO)-1,NADPH quinineoxidoreductase(NQO)-1,γ-glutamylcysteine synthetase(γ-GCS),runt-related transcription factor(RUNX)2 and bone morphogenic protein(BMP)2 mRNA levels in the aorta.Results The renal and aortic vascular structures of the control group rats were normal;the renal and aortic structures of the model group rats were severely damaged,and a large amount of crystalline adenine metabolites were deposited,and a continuous linear linear obvious calcified nodule was seen at the arterial media.Compared with the model group,the kidney lesions of rats in each TSRDN dose group and calcitriol group were significantly reduced,and adenine crystals we

关 键 词：穿山龙总皂苷 Nrf2/ARE通路 慢性肾脏病 血管钙化 

分 类 号：R285.5[医药卫生—中药学]