银杏叶提取物通过miR-145/FOXO1轴对实验性肾功能衰竭大鼠肾脏损伤的影响  

Effects of Ginkgo biloba extract on renal injury in rats with experimental renal failure through miR-145/FOXO1 axis

作　　者：李士旭 李林运 王新 李轲 卞华 LI Shixu;LI Linyun;WANG Xin;LI Ke;BIAN Hua(Department of Nephropathy and Rheumatism,Nanyang TCM Hospital,Nanyang 473000,Henan,China;College of Zhangzhongjing Traditional Chinese Medicine,Nanyang Institute of Technology,Nanyang 473004,Henan,China)

机构地区：[1]南阳市中医院肾病风湿科,河南南阳473000 [2]南阳理工学院张仲景国医国药学院,河南南阳473004

出　　处：《中国临床药理学与治疗学》2023年第7期728-735,共8页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：河南省中医药科学研究专项(2019ZY3028)。

摘　　要：目的:探讨银杏叶提取物(GBE)对慢性肾功能衰竭(CRF)大鼠肾损伤的影响及潜在的分子机制。方法:SD大鼠采用5/6肾切除术构建CRF模型,并分为模型(model)组、GBE组(100 mg/kg)、GBE+Agomir-NC组、GBE+Agomir-145组,每组12只;另取12只为假手术(sham)组,仅暴露肾脏,不进行肾脏切除。GBE组大鼠每日给予GBE100 mg/kg灌胃,1次/d,连续4周;GBE+AgomirNC组和GBE+Agomir-145组大鼠每日给予GBE100 mg/kg灌胃,然后每3 d一次分别通过尾静脉注射Agomir-NC和Agomir-145,连续4周;sham组和model组灌胃和尾静脉注射等量生理盐水。观察大鼠的一般状态,检测大鼠肾功能指标[24 h尿微量白蛋白(24 h UAlb)、血尿素氮(BUN)、血肌酐(SCr)]和氧化应激指标[丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSHPx)],采用Masson染色观察肾组织纤维化情况,采用实时荧光定量PCR(RT-qPCR)检测肾组织microRNA-145(miR-145)、转化生长因子-β1(TGF-β1)和叉头框转录因子O1(FOXO1)m RNA表达水平,采用Western blot检测肾组织TGF-β1、FOXO1蛋白水平。结果:GBE干预后CRF大鼠的一般状态明显好转,体质量、肾组织SOD和GSH-Px活性、FOXO1的mRNA和蛋白水平显著高于model组(P<0.05);24 h UAlb、血清BUN、SCr和肾组织MDA水平、肾间质纤维化的相对面积以及肾组织miR-145、TGF-β1的mRNA和蛋白水平均显著低于model组(P<0.05);且在GBE干预的基础上,上调miR-145表达可明显减弱GBE对CRF大鼠肾损伤的保护作用(P<0.05)。结论:GBE可减轻CRF大鼠的肾脏损伤,其作用机制可能与下调miR-145,上调FOXO1表达,抑制肾纤维化有关。AIM:To investigate the effect of Ginkgo biloba extract(GBE)on kidney injury in rats with chronic renal failure(CRF)and its potential molecular mechanism.METHODS:SD rats were given 5/6 nephrectomy to construct CRF models and divided into model group,GBE group(100 mg/kg),GBE+Agomir-NC group,and GBE+Agomir-145 group,12 per group;another 12 were selected as the sham group,with only the kidney exposed and no nephrectomy.Rats in the GBE group were given GBE 100 mg/kg gavage daily,once a day,for 4 consecutive weeks;rats in the GBE+Agomir-NC group and GBE+Agomir-145 group were given GBE 100 mg/kg gavage daily,and then Agomir-NC and Agomir-145 were injected via the tail vein every 3 days for 4 weeks;the sham group and the model group were given the same amount of normal saline by gavage and injection through the tail vein respectively.The general state of the rat was observed,and the renal function indicators[24 h urine microalbumin(24 h UAlb),blood urea nitrogen(BUN),blood creatinine(SCr)]and oxidative stress indicators[malonaldehyde(MDA),Superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)]were detected,Masson staining was used to observe the fibrosis of kidney tissue,real-time fluorescent quantitative PCR(RT-qPCR)was used to detect the mRNA expression levels of microRNA-145(miR-145),transforming growth factor-β1(TGF-β1)and forkhead box O1(FOXO1)in renal tissue,Western blot was used to detect the protein levels of TGF-β1 and FOXO1 in kidney tissue.RESULTS:The general state of CRF rats improved significantly after GBE intervention,the body weight,renal tissue SOD and GSH-Px activities,and FOXO1 mRNA and protein levels were significantly higher than those in the model group(P<0.05);the 24 h UAlb,serum BUN,SCr and renal tissue MDA levels,the relative area of renal interstitial fibrosis,and renal tissue miR-145,TGF-β1 mRNA and protein levels were significantly lower than those in the model group(P<0.05);and on the basis of GBE intervention,up-regulating the expression of miR-145 could significantly weaken the protec

关 键 词：银杏叶提取物 慢性肾功能衰竭 肾纤维化 microRNA-145 叉头框转录因子O1 

分 类 号：R965.2[医药卫生—药理学]