健脾补肾活血化浊方对糖尿病肾病小鼠FGF23/Pit⁃1信号通路及血管钙化的影响  

Effects of Jianpi Bushen Huoxue Huazhuo Prescription on FGF23/Pit⁃1 Signaling Pathway and Vascular Calcification in Diabetic Nephropathy Mice

作　　者：沈斌 朱国双 吴文静 王岚[2] SHEN Bin;ZHU Guo-shuang;WU Wen-jing;WANG Lan(Jiangsu Vocational College of Medicine,Yancheng 224005,China;Hubei University of Chinese Medicine,Wuhan 430061,China)

机构地区：[1]江苏医药职业学院,江苏盐城224005 [2]湖北中医药大学,湖北武汉430061

出　　处：《中药材》2020年第12期3005-3009,共5页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金青年项目(81703991)。

摘　　要：目的:探讨健脾补肾活血化浊方对糖尿病肾病(DN)小鼠成纤维细胞生长因子23(FGF23)/Ⅲ型钠磷协同转运子⁃1(Pit⁃1)信号通路及血管钙化的影响。方法:腹腔注射55 mg/kg链脲佐菌素构建DN小鼠模型,将50只造模成功小鼠随机分为5组,每组10只:模型组、厄贝沙坦(25 mg/kg)阳性对照组及健脾补肾活血化浊方低(1.3 g/kg)、中(2.6 g/kg)、高(5.2 g/kg)剂量组,并予以高磷饲料(含磷1.2%,钙1.6%);另选取10只正常小鼠设为正常对照组,各组动物灌胃给药,1次/d,,连续12 w,正常对照组和模型组灌胃等体积(10 mL/kg)蒸馏水。全自动生化分析仪检测各组小鼠空腹血糖(FBG)、24 h尿微量白蛋白(mALB)、钙、磷、尿素氮、血肌酐等生化指标;HE染色观察肾脏组织病理学变化;ELISA检测血清中FGF23表达;Western blot检测肾脏组织中α⁃Klotho(Kl)、Pit⁃1、核心结合因子α⁃l(Cbfα⁃l)蛋白表达。结果:治疗12 w后,与正常对照组比较,模型组小鼠肾小球体积增大、肾间质和肾小管纤维基底膜增厚、系膜基质增多,FBG、尿液中24 h mALB、血清磷、血肌酐、FGF23水平及肾脏组织中Pit⁃1、Cbfα⁃l蛋白表达显著升高(P<0.05),血清钙水平及肾脏组织中Kl蛋白表达显著降低(P<0.05);与模型组比较,各给药组小鼠肾组织病理损伤明显改善,FBG、尿液中24 h mALB、血清磷、血肌酐、FGF23水平及肾脏组织中Pit⁃1、Cbfα⁃l蛋白表达显著降低(P<0.05),血清钙及肾脏组织中Kl蛋白表达显著升高(P<0.05)及高剂量健脾补肾活血化浊方作用与厄贝沙坦相当。结论:健脾补肾活血化浊方可能通过抑制FGF23/Pit⁃1信号通路减轻血管钙化程度。Objective:To investigate the effects of Jianpi bushen huoxue huazhuo prescription on the fibroblast growth factor 23(FGF23)/typeⅢsodium⁃dependent phosphate cotransporter⁃1(Pit⁃1)signaling pathway and vascular calcification in diabetic nephrop⁃athy(DN)mice.Methods:The model of DN mice was established by intraperitoneal injection of 55 mg/kg streptozotocin,50 mice suc⁃cessfully modeled were randomly divided into 5 groups,10 in each group,including model group,positive control group of irbesartan(25 mg/kg)and low(1.3 g/kg),medium(2.6 g/kg),high(5.2 g/kg)dose groups of Jianpi bushen huoxue huazhuo prescription,fed with high phosphorus feed(contained 1.2%phosphorus,1.6%calcium);In addition,10 normal mice were selected as the normal control group.The mice in each group were given the drug by intragastric administration,once per day,for a continuous 12 weeks.The distilled water of the same volume(10 mL/kg)was intragastric administration in the normal control group and model group.Automatic biochemi⁃cal analyzer was used to detect fasting blood glucose(FBG),microalbumin(mALB)in urinary for 24 h,calcium,phosphorus,urea nitro⁃gen,blood creatinine and other biochemical indexes in each group;Histopathological changes of kidney were observed by HE stai⁃ning.The expression of FGF23 in serum was detected by ELISA.Western blot was used to detected the expressions ofα⁃Klotho(Kl),Pit⁃1 and core binding factorα⁃l(Cbfα⁃l)in kidney tissue.Results:After 12 weeks of treatment,compared with the normal control group,the volume of glomeruli in the mice of model control group increased,the basement membrane of renal interstitium and renal tu⁃bules was thickened,the mesangial matrix increased,the levels of FBG,mALB in urine for 24 h,phosphorus,creatinine,FGF23 in ser⁃um,Pit⁃1 and Cbfα⁃l proteins expressions in kidney tissues were significantly increased(P<0.05),and the level of calcium in serum and the protein expression of Kl in kidney tissues were significantly decreased(P<0.05);Compared with the mode

关 键 词：健脾补肾活血化浊方 糖尿病肾病 成纤维细胞生长因子23 Ⅲ型钠磷协同转运子⁃1 血管钙化 

分 类 号：R285.5[医药卫生—中药学]