四君子汤对慢性肾脏病-蛋白质能量消耗模型小鼠的改善作用及机制研究  

Study on the Improvement Effects of Sijunzi Decoction on Chronic Kidney Disease-protein Energy Wasting Model Mice and Its Mecranism

作　　者：许烨[1] 李志明[1] 远方[1] XU Ye;LI Zhiming;YUAN Fang(Dept.of Nephrology,the Affiliated Hospital of Liaoning University of TCM,Shenyang 110032,China)

机构地区：[1]辽宁中医药大学附属医院肾内科,沈阳110032

出　　处：《中国药房》2020年第19期2358-2364,共7页China Pharmacy

基　　金：辽宁省科学技术计划项目(No.20170540607)。

摘　　要：目的:研究四君子汤对慢性肾脏病-蛋白质能量消耗(CKD-PEW)模型小鼠骨骼肌萎缩的改善作用,并探讨其可能的作用机制。方法:将80只小鼠随机分为假手术组(n=10)和造模组(n=70)。造模组小鼠采用切除5/6肾联合低蛋白(4%酪蛋白)饮食法建立CKD-PEW模型。将造模成功的50只小鼠随机分为模型组,四君子汤低、中、高剂量组[2.34、4.68、9.36 g/(kg·d),以生药量计]和复方α-酮酸片组[阳性对照,1 g/(kg·d)],每组10只。各给药组小鼠灌胃相应药物,假手术组和模型组小鼠灌胃等体积生理盐水,每天1次,连续灌胃14 d。末次给药后,称定小鼠体质量、左侧胫骨前肌(TA)湿质量,并测定TA横截面积;检测小鼠TA蛋白合成与分解代谢情况;采用实时荧光定量-聚合酶链式反应法检测小鼠TA中B淋巴细胞瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)及胱天蛋白酶3(Caspase-3)mRNA表达水平;采用Western blotting法检测小鼠TA中肌萎缩蛋白Fbox-1(Atrogin-1)、肌环指蛋白1(MuRF-1)、Rho相关蛋白激酶1(ROCK1)、磷酸化肿瘤抑制基因(p-PTEN)、磷脂酰肌醇3激酶(PI3K)及磷酸化蛋白激酶B(p-Akt)蛋白的表达水平。结果:与假手术组比较,模型组小鼠的体质量、TA湿质量、TA蛋白合成代谢能力以及PI3K、p-Akt蛋白表达水平显著降低(P<0.05),TA横截面积显著减小(P<0.05),TA蛋白分解代谢能力、Bax/Bcl-2比值、Caspase-3 mRNA表达水平和Atrogin-1、MuRF-1、ROCK1、p-PTEN蛋白表达水平显著升高(P<0.05)。与模型组比较,四君子汤中、高剂量组和复方α-酮酸片组小鼠上述指标均显著改善(P<0.05)。结论:四君子汤能够增加CKD-PEW模型小鼠体质量,抑制其骨骼肌萎缩;其机制可能与调控ROCK1/PTEN/Akt信号通路,抑制Atrogin-1和MuRF-1表达有关。OBJECTIVE:To study the improvement effects of Sijunzi decoction on skeletal muscle atrophy in chronic kidney disease-protein energy wasting(CKD-PEW)model mice,and to explore its potential mechanism.METHODS:A total of 80 mice were randomly divided into sham operation group(n=10)and modeling group(n=70).CKD-PEW model was established by removing 5/6 kidneys and giving a low-protein diet(4%casein)for mice in modeling group.Totally 50 modeled mice were randomly divided into model group,Sijunzi decoction low-dose,medium-dose and high-dose groups[2.34,4.68,9.36 g/(kg·d),by crude drug],Compoundα-ketoacid tablets group[positive control,1 g/(kg·d)],with 10 mice in each group.Administration groups were given relevant medicine intragastrically;sham operation group and model group were given constant volume of normal saline intragastrically,once a day,for consecutive 14 d.After last medication,body weight of mice and wet mass of left tibialis anterior muscle(TA)were weighed;TA cross-sectional area was determined;protein synthesis and decomposition metabolism ability of TA were detected;m RNA expressions of Bcl-2,Bax and Caspase-3 in TA were detected by Real-time PCR;protein expressions of muscular dystrophin Fbox-1(Atrogin-1),myofloin-1(Mu RF-1),Rho-related protein kinase 1(ROCK1),phosphorylated PTEN(p-PTEN),phosphatidylinositol-3-kinase(PI3 K)and phosphorylated Akt(p-Akt)in TA were detected by Western blotting.RESULTS:Compared with the sham operation group,the body weight,TA wet weight,protein synthesis metabolism ability of TA as well as protein expressions of PI3 K and p-Akt were decreased significantly in model group(P<0.05);the cross-sectional area of TA decreased significantly(P<0.05);protein decomposition metabolism ability of TA,Bax/Bcl-2 ratio,Caspase-3 m RNA expression,protein expressions of Atrogin-1,Mu RF-1,ROCK1 and p-PTEN were increased significantly(P<0.05).Compared with model group,above indexes of mice were all improved significantly in Sijunzi decoction medium-dose,highdose groups and Compoundα-ketoacid

关 键 词：四君子汤 慢性肾脏病 蛋白质能量消耗 Rho相关蛋白激酶1 磷酸化肿瘤抑制基因 磷脂酰肌醇-3-激酶 蛋白激酶B 小鼠 

分 类 号：R285[医药卫生—中药学]