基于少阴枢机理论组方对膜性肾病小鼠肾组织HO-1蛋白表达的影响

Effect of prescription based on shaoyin pivot theory on HO-1 protein expression in renal tissue of mice with membranous nephropathy

关键词:  少阴枢机  麻黄附子细辛汤  防己黄芪汤  膜性肾病  HO-1

Key words:  shaoyin pivot  Mahuang Fuzi Xixin Decoction  Fangji Huangqi Decoction  membranous nephropathy  HO-1

摘要:

[目的] 探讨基于少阴枢机理论组方麻黄附子细辛汤合防己黄芪汤对膜性肾病小鼠肾组织的保护作用及其对HO-1蛋白表达的影响。[方法] 选用BALB/c小鼠50只，随机分为正常组10只、造模组40只。造模组小鼠尾静脉注射阳离子化牛血清白蛋白（C-BSA）制备膜性肾病模型，造模成功后，将造模小鼠采用随机数字表法分为4组，即模型组、中药组、西药组、中药加西药组。分别给予相应药物灌胃6周。6周后腹腔注射麻醉取材，摘眼球取血，检测各组小鼠血清总蛋白（TP）、三酰甘油（TG）、血肌酐（SCr）等指标水平；光镜下观察小鼠肾组织病理形态变化：苏木素-伊红（HE）染色，过碘酸-六胺银（PASM）染色；免疫荧光检测小鼠肾组织免疫球蛋白G（IgG）沉积；使用试剂盒检测小鼠肾组织丙二醛（MDA）含量，蛋白免疫印迹法（Western blot）检测小鼠肾组织HO-1蛋白表达情况。[结果] 与模型组比较，各给药组TP水平均升高，TG水平均降低（P＜0.01，P＜0.05）。各给药组之间，中药组与西药组比较，TP、TG水平差异无统计学意义（P＞0.05），与中药组及西药组比较，中药加西药组TP水平升高（P＜0.01，P＜0.05），TG水平降低（P＜0.05）。与模型组比较，中药组及西药组小鼠血清SCr水平差异无统计学意义（P＞0.05），中药加西药组差异有统计学意义（P＜0.05）。小鼠肾脏病理组织结构均有所改善。与模型组比较，各给药组小鼠肾组织MDA含量均降低（P＜0.05，P＜0.01）；各给药组之间，中药组与西药组比较，小鼠肾组织MDA含量差异不具有统计学意义（P＞0.05），与中药组及西药组比较，中药加西药组小鼠肾组织MDA含量降低（P＜0.05）。与模型组比较，各给药组小鼠肾组织中HO-1蛋白表达量均升高（P＜0.05，P＜0.01）；各给药组之间，中药组与西药组相比，小鼠肾组织中HO-1蛋白表达量差异无统计学意义（P＞0.05），与中药组比较，中药加西药组小鼠肾组织中HO-1蛋白表达量升高（P＜0.05）。[结论] 基于少阴枢机组方麻黄附子细辛汤合防己黄芪汤对膜性肾病小鼠的肾脏具有保护作用，其机制可能与上调HO-1蛋白的表达量有关。

Abstract:

[Objective] To investigate the protective effect of Mahuang Fuzi Xixin Decoction combined with Fangji Huangqi Decoction on renal tissue of mice with membranous nephropathy and the effect on the expression of HO-1 protein based on shaoyin pivot theory. [Methods] Fifty BALB/c mice were randomly divided into normal group（10 mice） and modeling group（40 mice）. Mice in the modeling group were injected cationized bovine serum albumin（C-BSA） into the tail vein to prepare membranous nephropathy model. After successful modeling，the mice were divided into four groups by random number table method：model group，traditional Chinese medicine（TCM） group，Western medicine group，and TCM plus Western medicine group. They were given the corresponding drugs intragastric administration for 6 weeks. Six weeks later，intraperitoneal injection was used for anesthesia，and blood was collected from eyeballs. Serum total protein（TP），triglyceride（TG），serum creatinine（SCr） and other indexes of each group were detected. Pathological changes of mouse kidney tissue were observed under light microscope：hematoxylin-eosin（HE） staining，silver periodate-hexamide（PASM） staining；immunofluorescence was used to detect the deposition of immunoglobulin G（IgG） in mouse kidney tissue. The content of malondialdehyde（MDA） in mouse kidney tissue was detected by the kit，and the expression of HO-1 protein in mouse kidney tissue was detected by Western blot. [Results] Compared with model group，TP level was increased and TG level was decreased in all drug administration groups（P＜0.01，P＜0.05）. Among administration groups，there was no statistical significance in TP and TG levels between TCM group and Western medicine group（P＞0.05），and compared with TCM group and the Western medicine group，the level of TP in TCM plus Westerm medicine group increased（P＜0.01，P＜0.05） and the level of TG decreased（P＜0.05）. Compared with model group，there was no statistical significance in serum SCr level between TCM group and Western medicine group（P＞0.05），but there was statistical significance in TCM plus Western medicine group（P＜0.05）. The pathological structure of mouse kidney was improved. Compared with model group，MDA content in kidney tissue of mice in each administration group was decreased（P＜0.05，P＜0.01）. There was no statistical significance in MDA content in renal tissue of mice between the TCM group and the Western medicine group（P＞0.05），but MDA content in renal tissue of mice in the TCM plus Western medicine group was decreased compared with the TCM group and the Western medicine group（P＜0.05）. Compared with model group，the expression level of HO-1 protein in kidney tissue of mice in each administration group was increased（P＜0.05，P＜0.01）. There was no significant difference in HO-1 protein expression level between the TCM group and the Western medicine group（P＞0.05），but the HO-1 protein expression level in the TCM plus Western medicine group was increased compared with the TCM group（P＜0.05）. [Conclusion] Based on shaoyin pivot theory，Mahuang Fuzi Xixin Decoction combined with Fangji Huangqi Decoction can protect the kidney of mice with membranous nephropathy，and the mechanism may be related to the up-regulation of HO-1 protein expression.