Renoprotective Effect of Sanjiao Qushi Prescription Against Cationic Bovine Serum Albumin Induced Membranous Nephropathy Mice Model and Its Influence on Nrf2/HO-1 Signaling Pathway

Ya-yun ZHAO; Jing FANG; Jing-jie CHEN; Hai-ping LIU; Zhi-qiang CHEN

doi:10.13422/j.cnki.syfjx.20211839

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Renoprotective Effect of Sanjiao Qushi Prescription Against Cationic Bovine Serum Albumin Induced Membranous Nephropathy Mice Model and Its Influence on Nrf2/HO-1 Signaling Pathway

Pharmacology | 更新时间：2021-08-18

- Renoprotective Effect of Sanjiao Qushi Prescription Against Cationic Bovine Serum Albumin Induced Membranous Nephropathy Mice Model and Its Influence on Nrf2/HO-1 Signaling Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 27, Issue 18, Pages: 58-65(2021)
- 作者机构：
  
  1.河北中医学院，石家庄 050091
  2.河北省中医院，石家庄 050011
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20211839
  CLC： R2-0;R334+.1;R33
- Received：28 June 2021，
  
  Published Online：28 July 2021，
  
  Published：20 September 2021
- 稿件说明：
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赵亚云,方敬,陈静洁等.三焦祛湿方对C-BSA诱导膜性肾病小鼠肾脏保护作用及其对Nrf2/HO-1信号通路的影响[J].中国实验方剂学杂志,2021,27(18):58-65.

ZHAO Ya-yun,FANG Jing,CHEN Jing-jie,et al.Renoprotective Effect of Sanjiao Qushi Prescription Against Cationic Bovine Serum Albumin Induced Membranous Nephropathy Mice Model and Its Influence on Nrf2/HO-1 Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(18):58-65.
赵亚云,方敬,陈静洁等.三焦祛湿方对C-BSA诱导膜性肾病小鼠肾脏保护作用及其对Nrf2/HO-1信号通路的影响[J].中国实验方剂学杂志,2021,27(18):58-65. DOI： 10.13422/j.cnki.syfjx.20211839.

ZHAO Ya-yun,FANG Jing,CHEN Jing-jie,et al.Renoprotective Effect of Sanjiao Qushi Prescription Against Cationic Bovine Serum Albumin Induced Membranous Nephropathy Mice Model and Its Influence on Nrf2/HO-1 Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(18):58-65. DOI： 10.13422/j.cnki.syfjx.20211839.

摘要

目的

探索三焦祛湿方对阳离子化牛血清白蛋白（C-BSA）诱导膜性肾病（MN）小鼠肾脏保护作用及其对核因子E

相关因子2（Nrf2）/血红素氧合酶-1（HO-1）信号通路影响。

方法

雌性BALB/c小鼠60只，随机分为正常组10只，模型组50只。模型组小鼠尾静脉注射C-BSA（6.5 mg·kg

-1

），造模成功小鼠按体质量随机分为模型组、三焦祛湿方低剂量（3.71 g·kg

-1

）组，三焦祛湿方高剂量（7.42 g·kg

-1

）组及盐酸贝那普利（1.3 mg·kg

-1

）组，分别给予各组相应剂量的药物灌胃，1次/d，连续4周。分别于造模前、造模后及治疗后检测24 h尿蛋白定量。给药后，腹主动脉取血检测血尿素氮（BUN），血清肌酐（SCr），甘油三酯（TG），总胆固醇（TC），总蛋白（TP），白蛋白（Alb）水平，肾组织行苏木素-伊红（HE），马松（Masson），过碘酸-六胺银（PASM）染色，光镜下观察小鼠肾组织病理形态变化，免疫荧光检测肾组织免疫球蛋白G（IgG）沉积情况，采用免疫荧光法检测肾组织中活性氧（ROS）表达情况。蛋白免疫印迹法（Western blot）观察肾组织Nrf2蛋白在细胞质和细胞核的表达情况，以及其下游因子血红素氧合酶-1（HO-1），醌还原型辅酶Ⅰ（NADH）脱氢酶1（NQO1）蛋白表达水平。

结果

模型组小鼠24 h尿蛋白水平显著升高（

0.01），血清TG，TC水平显著升高（

0.01），TP，Alb水平显著降低（

0.01）；光镜下观察肾脏病理改变，可见肾小球体积增大，基底膜增厚，嗜复红蛋白沉积，“钉突”形成；荧光显微镜下可见IgG沿毛细血管襻弥漫性沉积；肾组织ROS表达水平显著升高（

0.01）；肾组织细胞核内Nrf2蛋白表达显著升高（

0.01），细胞质Nrf2蛋白表达显著降低（

0.01），HO-1和NQO1蛋白表达显著升高（

0.01）；给药后，各治疗组小鼠24 h尿蛋白水平显著降低（

0.01），血清TG，TC水平显著降低（

0.01），TP，Alb水平明显升高（

0.05，

0.01）；各治疗组肾组织病理损害明显改善；肾组织ROS表达水平显著降低（

0.01）；肾组织细胞核内Nrf2蛋白表达显著降低（

0.01），细胞质Nrf2蛋白表达不同程度升高（

0.05，

0.01），HO-1和NQO1蛋白表达显著升高（

0.01）。

结论

三焦祛湿方可能通过调控膜性肾病肾组织Nrf2/HO-1信号表达，缓解肾组织氧化应激，进而降低24 h尿蛋白定量，降低血脂水平，提高血清蛋白含量，减轻肾脏病理损害，起到明显的肾脏保护及延缓病程进展的作用。

Abstract

Objective

To investigate the renoprotective effects that Sanjiao Qushi prescription ameliorates cationic bovine serum albumin （C-BSA） induced membranous nephropathy（MN） in mice model and its influence on nuclear factor erythroid-2-related factor-2（Nrf2）/heme oxygenase-1（HO-1） signaling pathway.

Method

Sixty female BALB/c mice were randomly divided into the normal group（

=10） and the model group（

=50）. The mice in the model group received C-BSA injection via tail vein （6.5 mg·kg

-1

）. The mice that were successfully modeled were randomized into the model group， the low dose Sanjiao Qushi prescription group（3.71 g·kg

-1

）， the high dose Sanjiao Qushi prescription group（7.42 g·kg

-1

） and benazepril hydrochloride group（1.3 mg·kg

-1

）. And they were administered with the corresponding medicine by gavage once a day for four consecutive weeks. 24 hour-urine protein quantitation were performed before C-BSA injection and after C-BSA injection as well as the medicine gavage. When the treatment was finished， all of the mice were sacrificed and the biochemical indicators such as serum creatinine（SCr）， blood urea nitrogen（BUN）， triglyceride（TG）， total cholesterol（TC）， total protein（TP） and albumin（Alb） were measured. And the renal pathological morphology changes were observed by light microscope with hematoxylineosin（HE）， Masson and periodic acid-silver metheramine（PASM） staining. The deposition of immunoglobulin G（IgG） in the glomerulus was detected by fluorescence microscope. The expression of reactive oxygen species （ROS） of kidney was detected by fluorescence immunoassay. The protein expression levels of Nrf2 in cell nucleus and cytoplasm and the downstream protein factors HO-1 and NADH quinone acceptor oxidoreductase 1（NQO1） were detected by Western blot.

Result

Compare to normal group， the levels of 24 hour-urine protein quantitation， TG and TC significantly increased in model group（

0.01）， while TP and Alb levels significantly decreased（

0.01）. The model group exhibited enlarged volume of glomerular， significantly thickened glomerular basement membrane（GBM）， fuchsinophilic protein deposition and spike formation through light microscope. Immunofluorescence staining for the model group exhibited granular deposition of IgG along the capillary wall. The expression of ROS in kidney significantly increased（

0.01）. The protein expression levels of Nrf2 in cell nucleus significantly increased（

0.01）， while Nrf2 in cytoplasm significantly decreased（

0.01）.The protein expression levels of HO-1 and NQO1 significantly increased（

0.01）. Compared to model group， the levels of 24 hour-urine protein quantitation， TG and TC significantly decreased in each treated group（

0.01）， TP and Alb levels significantly increased（

0.05，

0.01）. The pathological damages alleviated obviously. The expression of ROS in kidney significantly decreased（

0.01）. The protein expression levels of Nrf2 in cell nucleus significantly decreased（

0.01）， while Nrf2 in cytoplasm significantly increased（

0.05，

0.01）. The protein expression levels of HO-1 and NQO1 significantly increased（

0.01）.

Conclusion

Sanjiao Qushi prescription worked on MN mice possibly by regulating related proteins in the Nrf2/HO-1 signaling pathway and relieving oxidative stress， thus decreasing 24 hour-urine protein and blood lipid， increasing serum protein， and alleviating the pathological damages to protect renal function and delay progress of the disease.

关键词

Keywords

references

THOMPSON A ， CATTRAN D C ， BLANK M ， et al . Complete and partial remission as surrogate end points in membranous nephropathy ［J］. Jam Soc Nephrol ， 2015 ， 26 （ 12 ）： 2930 - 2937 ．

HOU J H ， ZHU H X ， ZHOU M L ， et al . Changes in the spectrum of kidney diseases：an analysis of 40759 biopsy-proven cases from 2003 to 2014 in China ［J］. Kidney Dis ， 2018 ， 4 （ 1 ）： 10 - 19 .

NANGAKU M ， SHANKLAND S J ， COUSER W G . Cellular response to injury in membranous nephropathy ［J］. J Am Soc Nephrol ， 2005 ， 16 （ 5 ）： 1195 - 204 .

KEUM Y S ， YU S ， CHANG P P ， et al . Mechanism of action of sul-foraphane： inhibition of p38 mitogen-activated protein kinase isoformscontributing to the induction of antioxidant response element-mediatedheme oxygenase-1 in human hepatoma HepG2 cells ［J］. Cancer Res ， 2006 ， 66 （ 17 ）： 8804 - 8813 .

秦卫松，刘志红，曾彩虹，等 . 雷公藤甲素对Heymann肾炎模型足细胞病变的影响［J］. 肾脏病与透析肾移植杂志， 2007 ， 6 （ 2 ）： 101 - 109 .

陈文军，陈素枝，靳晓华，等 . 降脂通络软胶囊对膜性肾病大鼠肾保护作用及对肾组织Bcl-2和Bad表达的影响［J］. 中草药， 2016 ， 47 （ 18 ）： 3263 - 3268 .

汪保和，皮亦华，黄畅，等 . 补肾通络方对阳离子化牛血清白蛋白致膜性肾病大鼠高凝状态的影响［J］. 中草药， 2009 ， 40 （ 7 ）： 1095 - 1098 .

黄谷香，刘瑞洪 . 重楼对膜性肾病大鼠肾脏的保护作用［J］. 广东医学， 2007 ， 28 （ 4 ）： 521 - 529 .

尼日特，杨巧芳，薛昕，等 . 加味过敏煎拆方组分对膜性肾病大鼠白细胞介素-4、白细胞介素-10等细胞因子的影响［J］. 中华中医药杂志， 2016 ， 31 （ 12 ）： 5277 - 5280 .

张芬芳，艾雨，陈志强，等 . 宣通三焦、活血通络方治疗特发性膜性肾病的临床疗效观察［J］. 中华中医药杂志， 2020 ， 35 （ 3 ）： 1596 - 1598 .

WU C C ， CHEN J S ， LIN S H ， et al . Experimental model of membranous nephropathy in mice：sequence of histological and biochemical events ［J］. Lab Anim ， 2008 ， 42 （ 3 ）： 350 - 359 .

秦川 . 实验动物学［M］. 北京：人民卫生出版社， 2010 ： 419 .

刘璐 . 宣通三焦、活血通络方对膜性肾病大鼠肾脏的保护作用及对肾组织中CD2AP和Podocin的影响［D］. 石家庄：河北医科大学， 2017 .

邹万忠 . 肾活检病理诊断标准指导意见［J］. 中华肾脏病杂志， 2001 （ 4 ）： 270 - 275 .

XU X ， WANG G ， CHEN N ， et al . Long⁃term exposure to air pollution and increased risk of membranous nephropathy in China ［J］. J Am Soc Nephrol ， 2016 ， 27 （ 12 ）： 3739 - 3746 .

TAKANO T ， ELIMAM H ， CYBULSKY A V . Complement-mediated cellular injury ［J］. Semin Nephrol ， 2013 ， 33 （ 6 ）： 586 - 601 ．

WHITE K E ， BILOUS R W ， MARSHALL S M ， et al . Podocyte number in normotensive type 1 diabetic patients with albuminuria ［J］. Diabetes ， 2002 ， 51 （ 10 ）： 3083 - 3089 ．

KERJASCHKI D ， NEALE T J . Molecular mechanisms of glomerular injury in rat experimental membranous nephropathy（Heymann nephritis）［J］. Am Soc Nephrol ， 1996 ， 7 （ 12 ）： 2518 - 2526 ．

ZHANG J ， WANG X ， VIKASH V ， et al . ROS and ROS-mediated cellular signaling ［J］. Oxid Med Cell Longev ， 2016 ， 2016 ： 4350965 .

KOBAYASHI A ， OHTA T ， YAMAMOTO M . Unique function of the Nrf2-Keap1 pathway in the inducible expression of antioxidant and detoxifying enzymes ［J］. Methods Enzymol ， 2004 ， 378 ： 273 - 286 .

MCCUBREY J A ， LAHAIR M M ， FRANKLIN R A . Reactive oxygen species-induced activation of the MAP kinase signaling pathways ［J］. Antioxid Redox Signal ， 2006 ， 8 （ 9/10 ）： 17 - 89 .

CAMP N D ， JAMES R G ， DAWSON D W ， et al . Wilms tumor gene on X chromosome （WTX） inhibits degradation of NRF2 protein through competitive binding to KEAP1 protein ［J］. J Biol Chem ， 2012 ，24， 287 （ 9 ）： 6539 - 6550 .

SO H ， KIM H ， KIM Y ， et al . Evidence that cisplatin-induced auditory damage is attenuated by downregulation of pro-inflammatory cytokines via Nrf2/HO-1 ［J］. J Assoc Res Otolaryngol ， 2008 ， 9 （ 3 ）： 290 - 306 .

KENSLER T W ， WAKABAYASHI N ， BISWAL S ， et al . Cell survival responses to environmental stresses via the Keap1-Nrf2-ARE pathway ［J］. Annu Rev Pharmacol ， 2007 ， 47 （ 1 ）： 89 - 116 .

YE T ， ZHEN J ， DU Y ， et al . Green tea polyphenol（-）-epigallocatechin-3-gallate restores Nrf2 activity and ameliorates crescentic glomerulonephritis ［J］. PLoS One ， 2015 ， 10 （ 3 ）： e0119543 .

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