均匀线性设计筛选Wilson病模型TX小鼠肾保护效应的实验研究

Experimental Study on Screening Renal Protective Effects for TX Mice Models with Wilson Disease Based on Uniform Linear Design

目的:筛选对于TX小鼠肾脏起保护作用的效应中药方剂肝豆汤。方法:按照均匀设计法6因素11水平制备实验中药;采用原子吸收法、Western blot法检测不同药物组方对TX小鼠肾脏组织微量元素铜(Gu)、铁(Fe)、锌(Zn)及组织蛋白B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associatedX protein,Bax)、半胱氨酸天冬氨酸蛋白酶-3(Cysteinyl aspartate-specific proproteases-3,Caspase-3)表达的影响,运用SPSS 17.0软件进行多元回归分析并筛选对TX小鼠肾脏起保护作用的效应中药。结果:本实验筛选出肝豆汤中与肾组织Cu、Zn含量密切相关的效应中药有X_6(三七,应取最大值0.009 g入方,P=0.030),X_2(姜黄,应取最大值0.059 g入方,P=0.003);本实验未筛选出肝豆汤组方中与TX小鼠肾组织Fe含量相关的效应中药;肝豆汤中与TX小鼠肾组织凋亡蛋白Bcl-2、Bax、Caspase-3表达密切相关的效应中药有X3(黄连,应取最大值0.059 g入方,P=0.008)、X4(泽泻,应取最大值0.072g入方,P=0.014)、X5(金钱草,应取最大值0.072 g入方,P=0.008)、X_6(三七,应取最大值0.009 g入方,P=0.014)。结论:肝豆汤肾保护效应中药最佳配伍及剂量配比为:黄连0.059 g、姜黄0.059 g、金钱草0.072 g、泽泻0.072 g、三七0.009 g,大黄未进入上述指标回归方程。中药肝豆汤中黄连、姜黄、金钱草、泽泻、三七通过调节微量元素铜锌代谢及调控凋亡蛋白多靶点发挥对TX小鼠肾脏的保护作用,大黄对于TX小鼠的肾脏保护作用尚有待证实。

Objective:To screen GanDou decoction which could proted the kidney of TX mice.Methods:Herbs were prepared according to 11-level 6-factor in uniform design;the effects of different herbal prescriptions on the expressions of tissue proteins Bax,Bcl-2 and Caspase-3 and trace elements copper(Cu),ferrum(Fe)and Zinc(Zn)in renal tissue of TX mice were detected by atomic absorption method and Western blot method,multiple regression analysis was performed using SPASS software and the effector herbs which could protect the kidney of TX mice were chosen.Results:The herbs which were closely related tothe contents of Cu and Zn in renal tissues in GanDou decoction were selected in the experiment,and they were X6(SanQi,maximum 0.09 g should be chosen into the prescription,P=0.030),and X2(JiangHuang,maximum 0.059 should be chosen into the prescription,P=0.003);the herbs related to the contents of Fe in renal tissue of TX mice in the formation of GanDou decoction haven't been selected in the experiment;the herbs closely related to the expressions of apoptotic proteins Bax,Bcl-2 and Caspase-3 in renal tissues of TX mice in GanDou decoction were X3(HuangLian,maximum 0.059 g should be chosen into the prescription,P=0.008),X4(ZeXie,maximum 0.072g should be chosen into the prescription,P=0.014),X5(JinQianCao,maximum 0.072g should be chosen into the prescription,P=0.008),X6(SanQi,maximum 0.009g should be chosen into the prescription,P=0.014).Conclusion:The optimal compatibility and dose proportions of herbs which could protect kidney in GanDou decoction are:HuangLian 0.059g,JiangHuang 0.059g,JinQianCao 0.072g,ZeXie 0.072 g,SanQi 0.009g,and DaHuang doesn't enter in the regression equation of the above indexes.HuangLian,JiangHuang,JinQianCao,ZeXie and SanQi in GanDou decoction could develop the effects of protecting the kidney of TX mice by regulating the metabolism of trace elements Cu and Zn and apoptotic proteins multitargets,the protective effects of DaHuang to the kidney of TX mice need to be proved.