基于Wnt4/β-catenin信号通路探讨和肾络宁风颗粒对慢性肾衰竭钙磷代谢紊乱大鼠干预机制  

The Intervention Mechanism of Heshenluoningfeng Granule on the Disorder of Calcium and Phosphorus Metabolism in Chronic Renal Failure Basing by Wnt4/β-catenin signal pathway

作　　者：郝媌 安娜 于惠青 尹爱兵 任超 于俊生 Hao Miao;An Na;Yu Huiqing;Yin Aibing;Ren Chao;Yu Junsheng(Shandong Qingdao Hospital of Intergrated Traditional Western Medicine, Qingdao 266002;QingdaoUniversity of Basic Medicine Qingdao,266071;Qingdao Hiser Medical Group,Qingdao 266033)

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

出　　处：《中药药理与临床》2019年第1期143-149,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：青岛市中医药科研计划项目(2017-zyx025)

摘　　要：目的:观察和肾络宁风颗粒对慢性肾衰竭钙磷代谢紊乱模型大鼠Wnt4/β-catenin信号通路的干预作用,探讨可能的作用机制。方法:雄性Wistar大鼠用5/6肾切除及高磷饮食制备大鼠模型,持续灌胃给药8周。HE染色观察肾组织形态学变化;检测各组血清肌酐(Scr)、尿素氮(BUN)、24h尿蛋白定量、血钙(Ca^(2+))、血磷(P^(3+))、血清甲状旁腺激素(PTH)、血清碱性磷酸酶(ALP)。免疫组化和Rt-PCR法分别检测大鼠肾脏Wnt4、β-catenin蛋白和基因表达水平。结果:与正常组比较,模型组大鼠肾小球结构紊乱、系膜细胞增生,基质增多,肾小管代偿性扩张,间质有大量炎性细胞浸润及纤维组织增生,血Scr、BUN、P^(3+)、PTH、ALP水平明显升高,血Ca^(2+)水平下降,和肾络宁风颗粒16.2、32.4g/kg组肾小球增生硬化程度及肾间质炎性细胞浸润情况较模型组明显改善;血Scr、BUN、P^(3+)、PTH、ALP水平显著降低,血Ca^(2+)水平明显上升;和肾络宁风颗粒各组Wnt4、β-catenin蛋白表达与基因表达显著下调。与骨化三醇0.045ug/kg组比较,和肾络宁风颗粒16.2、32.4g/kg组血Scr、BUN和P^(3+)水平显著降低,Wnt4/β-catenin的蛋白和基因下调。结论:和肾络宁风颗粒能够改善慢性肾衰钙磷代谢紊乱大鼠肾功能和钙磷代谢,延缓病情进展,其作用机制可能与下调Wnt4/β-catenin信号通路有关,且在一定范围内呈剂量依赖性。Objective: To observe the effect of Heshenluoningfeng Granule on chronic renal failure( CRF) rats model with calcium and phosphorus metabolic disorders by regulating Wnt4-β-catenin signal pathway,and to explore its possible mechanism. Methods: CRF rats models was prepared by 5/6 nephrectomy and high-phosphorus diet. After the administration with corresponding drugs for 8 weeks,histological changes in kidney renal tissues were observed by HE staining,then levels of blood urea nitrogen( BUN),serumcreatinine( Scr),serum calcium( Ca2+),phosphorus( P3+),Alkaline phosphatase( ALP),Parathyroid hormone( PTH) and 24-hour urine protein were detected. Protein and gene expressions levels of Wnt4,β-catenin were detected by immunohistochemistry and real-time quantitative. Results : Compared with the normal group,disordered glomerular structure,hyperplastic mesangial cells,partial glomerulosclerosis,tubular ectasia,inflammatory cell infiltration and focal tissue hyperplasia in renal interstitium were observed in the model group. The degrees of glomerular hyperplasia and interstitial inflammatory cell infiltration in Heshenluoningfeng Granule were reduced than those in the model group. Compared with the normal group,serum levels of Scr,BUN,P3+,PTH and ALP were significantly increased,while serum Ca2+ level was decreased in the model group. Compared with the model group,serum levels of above indicators were significantly reduced,while serum Ca2+ level was increased in Heshenluoningfeng Granule group. Compared with the positive control group,serum levels of Scr,BUN and P3+ were further reduced,the protein and gene expressions of Wnt4 and β-catenin were down-regulated in Heshenluoningfeng Granule group. Conclusion: Heshenluoningfeng Granule can improve the renal function,calcium and phosphorus metabolism and delay the disease progress. Its mechanism may be related to down-regulating of Wnt4/β-catenin signaling pathway,in a dose-dependent manner.

关 键 词：和肾络宁风颗粒 慢性肾衰竭钙磷代谢紊乱 Wnt4/β-catenin信号通路 

分 类 号：R285.5[医药卫生—中药学]