当归红芪超滤膜提取物调控肾脏缺氧介导的HIF⁃1α信号通路改善DKD大鼠肾脏纤维化的机制  

Mechanism of ultrafiltration extract of Angelicae Sinensis Radix and Hedysari Radix regulates HIF⁃1αsignaling pathway mediated by renal hypoxia to ameliorate renal fibrosis in DKD rats

作　　者：张小琳 李荣科[1] 高婷 徐庆全 万生芳[1] 张磊[1] 卢添翼 ZHANG Xiao-lin;LI Rong-ke;GAO Ting;XU Qing-quan;WAN Sheng-fang;ZHANG Lei;LU Tian-yi(Gansu University of Chinese Medicine,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学,兰州730000

出　　处：《中国中药杂志》2024年第6期1602-1610,共9页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82060914);甘肃省中医药研究中心开放课题(zyzx⁃2020⁃zx11);甘肃中医药大学2022年度中医学一级学科“岐黄英才”导师专项基金硕士研究生导师开放基金项目(12);2021年甘肃省青年博士基金项目(2022QB⁃096)。

摘　　要：通过当归红芪超滤膜提取物干预糖尿病肾病(diabetic kidneydisease,DKD)模型大鼠,观察DKD大鼠肾脏缺氧介导的缺氧诱导因子⁃1α(hypoxia inducible factor⁃1α,HIF⁃1α)/血管内皮生长因子(vascular endothelial growth factor,VEGF)和HIF⁃1α/血小板源性生长因子(platelet⁃derived growth factor,PDGF)/血小板源性生长因子受体(platelet derived growth factor receptor,PDGFR)信号通路的表达,探讨当归红芪超滤膜提取物改善DKD大鼠肾脏纤维化的作用机制。将50只SPF级雄性Wistar大鼠适应性喂养1周后,随机分为空白组7只及造模组,造模组禁食24 h后一次性腹腔注射链脲佐菌素(streptozotocin,STZ)联合高糖高脂饲料喂养制备DKD模型,造模成功后,将造模组随机分为模型组,当归红芪超滤膜提取物低、中剂量组,各7只,厄贝沙坦组和当归红芪超滤膜提取物高剂量组各8只。给予药物干预12周,观察大鼠一般情况,每周检测大鼠的体质量及血糖,于灌胃第6、12周检测大鼠24 h尿蛋白(urinary protein,UP);末次给药后,检测大鼠肾功能相关指标;Masson染色观察肾组织病理变化;免疫组化法(immunohistochemistry,IHC)检测肾组织脯胺酰羟化酶2(prolyl hydroxylase domain 2,PHD2)、HIF⁃1α蛋白表达;实时荧光定量聚合酶链式反应(real⁃time qPCR)检测肾组织PHD2、VEGF、PDGF、PDGFR mRNA表达;蛋白免疫印迹法(Western blot)检测肾组织HIF⁃1α、VEGF、PDGF、PDGFR蛋白的表达。结果显示,与模型组比较,各给药组大鼠血清中糖化血清蛋白(glycosylated serum protein,GSP)、血肌酐(serum creatinine,Scr)、尿素氮(blood urea nitrogen,BUN)水平不同程度降低(P<0.05或P<0.01),呈剂量相关性;大鼠肾脏病理明显改善;大鼠肾脏中PHD2 mRNA表达显著上调(P<0.05或P<0.01),VEGF、PDGF、PDGFR mRNA表达显著下调(P<0.05或P<0.01);大鼠肾脏中HIF⁃1α、VEGF、PDGF、PDGFR蛋白表达水平显著降低(P<0.01);PHD2阳性细胞率表达上升(P<0.01)。综上,当归�This study explored the mechanism of the ultrafiltration extract of Angelicae Sinensis Radix and Hedysari Radix in ameliorating renal fibrosis in the rat model of diabetic kidney disease(DKD)based on the expression of hypoxia⁃inducible factor⁃1α(HIF⁃1α)/vascular endothelial growth factor(VEGF)and HIF⁃1α/platelet⁃derived growth factor(PDGF)/platelet⁃derived growth factor receptor(PDGFR)signaling pathways in the DKD rats.After 1 week of adaptive feeding,50 male SPF⁃grade Wistar rats were randomized into a blank group(n=7)and a modeling group.After 24 h of fasting,the rats in the modeling group were subjected to intraperitoneal injection of streptozocin and fed with a high⁃sugar and high⁃fat diet to establish a DKD model.After modeling,the rats were randomly assigned into model(n=7),low⁃dose ultrafiltration extract(n=7),medium⁃dose ultrafiltration extract(n=7),irbesartan(n=8),and high⁃dose ultrafiltration extract(n=8)groups.After intervention by corresponding drugs for 12 weeks,the general conditions of the rats were observed.The body weights and blood glucose levels of the rats were measured weekly,and the 24 h urinary protein(24hUP)was measured at the 6th and 12th weeks of drug administration.After the last drug administration,the renal function indicators were determined.Masson staining was employed to observe the pathological changes of the renal tissue.The expression of prolyl hydroxylase domain 2(PHD2)and HIF⁃1αin the renal tissue was detected by immunohistochemistry(IHC).Real⁃time qPCR was employed to determine the mRNA levels of PHD2,VEGF,PDGF,and PDGFR in the renal tissue.Western blot was employed to determine the protein levels of HIF⁃1α,VEGF,PDGF,and PDGFR in the renal tissue.The results showed that compared with the model group,drug administration lowered the levels of glycosylated serum protein(GSP),aerum creatinine(Scr),and blood urea nitrogen(BUN)in a dose⁃dependent manner(P<0.05 or P<0.01)and mitigated the pathological changes in the renal tissue.Furthermore,drug

关 键 词：当归红芪超滤膜提取物 糖尿病肾病 肾脏缺氧 细胞外基质沉积 HIF⁃1α信号通路 

分 类 号：R285.5[医药卫生—中药学]