基于Klotho-FGF23轴探讨加味六味地黄汤对CKD-MBD模型大鼠骨保护作用的机制  

Mechanism of Modified Liuwei Dihuangtang in Bone Protection of CKD-MBD Model Rats:An Exploration Based on Klotho-FGF23 Axis

作　　者：郭华慧 李美丹 黄仁发 梁群卿[3] 李贺生 刘雪频 王若琳 沈思亨 GUO Hua-hui;LI Mei-dan;HUANG Ren-fa;LIANG Qun-qing;LI He-sheng;LIU Xue-pin;WANG Ruo-lin;SHEN Si-heng(Shenzhen Hospital of Guangzhou University of Chinese Medicine(Futian),Shenzhen 518034,China;Zhaoqing Hospital of Traditional Chinese Medicine,Zhaoqing 525000,China;Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530011,China)

机构地区：[1]广州中医药大学深圳医院(福田),广东深圳518034 [2]肇庆市中医院,广东肇庆525000 [3]广西中医药大学附属瑞康医院,广西南宁530011

出　　处：《中国实验方剂学杂志》2021年第24期61-70,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81760805,81460682);广东省自然科学基金项目(2020A151501572);深圳市科技创新委员会课题项目(JCYJ20190809102413156)。

摘　　要：目的:观察加味六味地黄汤对高磷联合腺嘌呤大鼠血清成纤维细胞生长因子23(FGF23),全段甲状旁腺激素(iPTH)和1,25-二羟维生素D3[1,25(OH)2D3]水平及肾骨组织Klotho和FGF23蛋白表达的影响,探讨加味六味地黄汤调控Klotho-FGF23对治疗肾性骨病的作用机制。方法:健康成年SD大鼠130只,随机分为正常组(n=10),高磷组(n=30),模型组(n=30),加味六味地黄汤组(n=30)和骨化三醇组(n=30)5组,各组组内分8,10,12周3个时间点。正常组大鼠普通饮食饲养,高磷组高磷饮食饲养,其他组大鼠腺嘌呤及高磷饮食饲养建立肾性骨病大鼠模型,正常组、高磷组、模型组大鼠均予以蒸馏水(10 mL·kg^(-1)·d^(-1))灌胃;加味六味地黄汤组予加味六味地黄汤药液(2.556 g·kg^(-1)·d^(-1))灌胃;骨化三醇组予骨化三醇(0.09μg·kg^(-1)·d^(-1))灌胃。结果:与正常组和高磷组8,10,12周比较,模型组大鼠血尿素氮(BUN),血肌酐(SCr),血磷,iPTH,FGF23,肾间质纤维化评分,肾骨组织FGF23蛋白均显著增高,血清钙水平,1,25(OH)2D3和肾、骨组织Klotho蛋白表达均降低(P<0.05,P<0.01);与模型组8,10,12周比较,加味六味地黄汤组和骨化三醇组大鼠BUN,SCr,血磷,iPTH,FGF23,小管间质半定量评分,肾骨组织FGF23蛋白表达均显著降低,血清钙水平,1,25(OH)2D3和肾、骨组织Klotho蛋白增高(P<0.05,P<0.01);而加味六味地黄汤组和骨化三醇组两组间BUN,SCr,血磷,iPTH,FGF23,肾间质纤维化评分,肾、骨组织FGF23蛋白表达差异无统计学意义。结论:Klotho-FGF23轴调控钙磷代谢可能参与了肾性骨病过程;加味六味地黄汤能有效改善肾功能、肾脏和骨组织病理,调节钙磷代谢紊乱,其骨保护作用机制与调控Klotho-FGF23轴有关。Objective:To observe the effects of modified Liuwei Dihuangtang on serum fibroblast growth factor 23(FGF23),full-length intact parathyroid hormone(iPTH),and 1,25-dihydroxyvitamin D3[1,25(OH)2 D3]levels and Klotho and FGF23 protein expression in renal and bone tissues of rats exposed to high phosphorus combined with adenine,so as to explore the mechanism of modified Liuwei Dihuangtang against renal osteopathy.Method:One hundred and thirty healthy adult SD rats were randomly divided into five groups,namely normal group(n=10),high phosphorus group(n=30),model group(n=30),modified Liuwei Dihuangtang group(n=30),and calcitriol group(n=30),and rats in each group were further classified based on three time points,namely 8,10,and 12 weeks.Rats in the normal group were fed with normal diet,the ones in the high phosphorus group with high phosphorus diet,and those in the other groups with adenine and high phosphorus diet for inducing renal osteopathy.Rats in the normal group,high phosphorus group,and model group were intragastrically administered with distilled water(10 mL·kg^(-1)·d^(-1)),the ones in the modified Liuwei Dihuangtang group with modified Liuwei Dihuangtang(2.556 g·kg^(-1)·d^(-1)),and those in the calcitriol group with calcitriol(0.09 μg·kg^(-1)·d^(-1)).Result:Compared with the normal group and high phosphorus group at the weeks of 8,10 and 12,the model group displayed significantly elevated blood urea nitrogen(BUN),serum creatinine(SCr),serum phosphorus,iPTH,FGF23,renal interstitial fibrosis score,and FGF23 expression in renal and bone tissues,but lowered serum calcium and 1,25(OH)2 D3 and Klotho protein expression in renal and bone tissues(P<0.05,P<0.01).Compared with the model group at the weeks of 8,10 and 12,the modified Liuwei Dihuangtang and calcitriol both significantly decreased the serum BUN,SCr,serum phosphorus,iPTH,FGF23,tubulointerstitial semi-quantitative score,and FGF23 expression in renal and bone tissues,while increased the serum calcium,1,25(OH)2 D3,and Klotho protein expression in rena

关 键 词：加味六味地黄汤 肾性骨病 成纤维细胞生长因子23 KLOTHO蛋白 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]