和肾络宁风颗粒干预FGF23/Klotho轴对慢性肾衰竭钙磷代谢紊乱模型大鼠的影响  

Effect of Heshenluo Ningfeng Granules on Calcium and Phosphorus Metabolism Disorder in Rats with Chronic Renal Failure by Intervening FGF23/Klotho Axis

作　　者：郝媌 安娜 于惠青 尹爱兵 于俊生 HAO Miao;AN Na;YU Huiqing;YIN Aibing;YU Junsheng(Shandong Qingdao Hospital of Integrated Traditional Western Medicine,Qingdao 266002,Shandong,China;Qingdao University of Basic Medicine,Qingdao 266071 Shandong,China;Qingdao Hiser Medical Group,Qingdao 266033 Shandong,China)

机构地区：[1]山东青岛中西医结合医院,山东青岛266002 [2]青岛大学基础医学院,山东青岛266071 [3]青岛市海慈医疗集团,山东青岛260033

出　　处：《中药新药与临床药理》2020年第5期530-538,共9页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：青岛市中医科研计划项目(2017-zyx025);青岛市2020年医疗卫生优秀人才培养项目(青卫科教字2019-09)。

摘　　要：目的观察和肾络宁风颗粒通过FGF23/Klotho轴对慢性肾衰竭钙磷代谢紊乱模型大鼠的干预机制。方法采用5/6肾切除及高磷饮食喂养构建慢性肾衰竭钙磷代谢紊乱大鼠模型,模型复制成功后给予和肾络宁风颗粒灌胃,持续给药8周。以PAS染色观察肾组织变化,检测血清肌酐(Scr)、尿素氮(BUN)、血钙(Ca2+)、血磷(P3+)、血清碱性磷酸酶(ALP)、血清甲状旁腺激素(PTH)等,免疫组化、Western Blot法和RT-PCR法检测大鼠肾脏Klotho和股骨FGF23的蛋白及基因表达。结果 1. PAS染色观察模型组大鼠肾小球结构紊乱、肾小管扩张,间质有大量炎性细胞浸润;与模型组比较,和肾络宁风颗粒中、高剂量组肾小球轻度增生,肾小管轻度扩张,间质炎性细胞浸润数目减少。2.与正常组比较,模型组大鼠血Scr、BUN、P3+、PTH、ALP明显升高,体质量和血Ca2+下降(P<0.01),Klotho蛋白及mRNA表达下调(P<0.05,P<0.01),FGF23蛋白及mRNA表达上调(P<0.01)。与模型组比,和肾络宁风颗粒中、高剂量组血Scr、BUN、P3+、PTH、ALP明显降低,体质量和血Ca2+升高(P<0.01),Klotho蛋白及mRNA表达上调(P<0.01),FGF23蛋白及mRNA表达下调(P<0.01)。与骨化三醇组比较,和肾络宁风颗粒中、高剂量组血Scr、BUN和P3+降低(P<0.01,P<0.05),FGF23蛋白和基因表达下降(P<0.01,P<0.05)。结论和肾络宁风颗粒可能通过对FGF23/Klotho轴的调控改善慢性肾衰钙磷代谢紊乱模型大鼠的肾功能和钙磷代谢,在一定范围内呈剂量依赖性。Objective To observe the intervention mechanism of Heshenluo Ningfeng Granule(HNG)on rat model of chronic renal failure induced with calcium and phosphorus metabolic disorders through the FGF23/Klotho axis.Methods The rats were subjected to 5/6 nephrectomy and fed with high-phosphorus diet to prepare the models.The rats were given 8 weeks drug intervention after models were successful established. Histological changes in renal tissues were observed by Periodic Acid-Schiff stain(PAS)staining. Blood urea nitrogen(BUN),serum creatinine(Scr),serum calcium(Ca2+),phosphorus(P3+),alkaline-phosphatase(ALP),parathyroid hormone(PTH)were detected by automatic biochemical analyzer and immunochemiluminometric assays. Expression of fibroblast growth factor-23(FGF23)and klotho in renal cortex and fresh femur were visualized by immunohistochemistry,and the protein and mRNA levels were further detected by Western Blot and real-time quantitative PCR. Results 1. PAS staining showed that compared with the normal group, disordered glomerular structure, tubular ectasia, and inflammatory cell infiltration were shown in the model group. The degree of glomerular hyperplasia and interstitial inflammatory cell infiltration in the medium and high dose groups of HNG were reduced compared with that in model group. 2. Compared with normal group,serum levels of Scr,BUN,P3+,PTH and ALP were significantly increased,the body weight and serum level of Ca2+decreased in the model group(P < 0.01);the expression levels of protein and mRNA of Klotho were decreased(P < 0.05,P < 0.01),and the expression levels of FGF23 increased(P <0.01). Compared with the model group,levels of all above indicators were significantly reduced,while the body weights and serum levels of Ca2+increased in the medium and high dose groups of HNG(P < 0.01). The expression levels of protein and mRNA of Klotho were increased(P < 0.01),and the expression levels of FGF23 decreased significantly(P < 0.01). Compared with the calcitriol group,serum levels of Scr,BUN and P3+were further

关 键 词：慢性肾衰竭钙磷代谢紊乱 和肾络宁风颗粒 KLOTHO蛋白 FGF23 肾功能 大鼠 

分 类 号：R285.5[医药卫生—中药学]