六味地黄丸干预FGF23/Klotho轴对慢性肾衰竭钙磷代谢紊乱模型大鼠的影响  

The Effect of Liuwei Dihuang Pill(六味地黄丸)on FGF23/Klotho Axis in Chronic Kidney Disease Model Rat with Calcium and Phosphorus Metabolism Disorder

作　　者：李旭华[1] 彭亚军[1] 盛望[1] 徐文峰[1] 刘丙龙 LI Xuhua;PENG Yajun;SHENG Wang;XU Wenfeng;LIU Binglong(The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha Hunan 410007,China;Hunan University of Chinese Medicine,Changsha Hunan 410208,China)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007 [2]湖南中医药大学,湖南长沙410208

出　　处：《中医药导报》2024年第10期36-40,59,共6页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：湖南省卫生健康委员会资助课题(202103052121);湖南省卫生健康委员会一般资助课题(202103050979);湖南省教育厅科学研究(优秀青年项目)资助项目(20B432);长沙市自然科学基金(kq2014220)。

摘　　要：目的:观察六味地黄丸通过成纤维细胞生长因子23(FGF23)/Klotho轴对慢性肾衰竭钙磷代谢紊乱模型大鼠的影响及其干预机制。方法:50只SPF级SD大鼠随机分为假手术组(10只)、手术组(40只),手术组采用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,模型复制成功后将30只造模成功大鼠随机分为模型组、骨化三醇组、六味地黄丸组,每组10只。六味地黄丸组、骨化三醇组大鼠分别予六味地黄丸和骨化三醇灌胃,假手术组及模型组大鼠予等体积的蒸馏水灌胃,持续给药8周。HE染色观察肾组织变化,检测大鼠血清钙(Ca^(2+))、磷(P^(3+))、碱性磷酸酶(ALP)、甲状旁腺激素(PTH)、肌酐(Scr)、尿素氮(BUN),免疫组化及Western blotting法检测大鼠肾皮质Klotho、FGF23蛋白的表达。结果:HE染色显示,模型组大鼠肾小球结构紊乱、肾小管扩张,间质有大量炎症细胞浸润;与模型组比较,六味地黄丸组大鼠肾小球轻度增生,肾小管轻度扩张,间质炎症细胞浸润数目减少。模型组大鼠血清Scr、BUN、P^(3+)、PTH、ALP高于假手术组(P<0.01),体质量、血清Ca^(2+)低于假手术组(P<0.01);六味地黄丸组大鼠血清Scr、BUN、P^(3+)、PTH、ALP低于模型组,体质量、血清Ca^(2+)高于模型组(P<0.01)。模型组大鼠肾皮质Klotho蛋白相对表达量低于假手术组(P<0.05),FGF23蛋白相对表达量高于假手术组(P<0.01);六味地黄丸组大鼠肾皮质Klotho蛋白相对表达量高于模型组(P<0.01),FGF23蛋白相对表达量低于模型组(P<0.01)。六味地黄丸组大鼠血Scr、BUN及肾皮质FGF23、Klotho蛋白相对表达量与骨化三醇组比较,差异无统计学意义(P>0.05)。六味地黄丸组大鼠血P^(3+)、Ca^(2+)高于骨化三醇组(P<0.05)。结论:六味地黄丸可能通过调控FGF23/Klotho轴改善慢性肾衰竭钙磷代谢紊乱模型大鼠的肾功能和钙磷代谢。Objective:To observe the intervention mechanism of Liuwei Dihaung pill in chronic kidney disease model rat with calcium and phosphorus metabolism disorder,through FGF23/Klotho axis.Methods:Totally 50 SPF SD rats were randomly divided into sham operation group(n=10)and operation group(n=40).The 5/6 nephrectomy and high phosphorus diet feeding as appliedto construct the rat model of chronic renal failure with calcium and phosphorus metabolism disorder in operation group.After successful model replication,30 rats were randomly divided into model group,calcitriol group and Liuwei Dihuang pill group,with 10 rats in each group.Rats were given Liuwei Dihuang pill and calciriol respectively in Liuwei Dihuang pill group and calcitriol group,while equal volume distilled water in sham operation group and model group for 8 weeks.Kidney tissue changes were observed by HE staining,and serum calcium(Ca^(2+)),serum phosphorus(P^(3+)),serum alkaline phosphatase(ALP),serum parathyroid hormone(PTH),creatinine(Scr),and urea nitrogen(BUN)were detected.The protein expression of Klotho and FGF 23 in renal cortex was detected by immunohistochemistry and Western blotting.Results:HE staining showed there were disturbed glomerular structure,tubular expansion and massive inflammatory cells in model group.Compared with the model group,there were mild hyperplasia and mild dilation of renal tubules in Liuwei Dihuang pill group,and the number of interstitial inflammatory cells decreased.The model group showed higher blood Scr,BUN,P^(3+),PTH and ALP than sham operation group(P<0.01),while lower body weight and blood Ca^(2+)than sham operation group(P<0.01).Liuwei Dihuang pill group showed lower blood Scr,BUN,P^(3+),PTH and ALP than model group(P<0.01),while higher body weight and blood Ca^(2+)than model group(P<0.01).The model group showed lower Klotho protein expression than sham operation group(P<0.05),while higher FGF23 protein expression than sham operation group(P<0.01).Liuwei Dihuang pill group showed higher Klotho protein expression than

关 键 词：慢性肾衰竭钙磷代谢紊乱 六味地黄丸 KLOTHO蛋白 FGF23蛋白 大鼠 

分 类 号：R285.5[医药卫生—中药学]