Effect and Mechanism of Baoshen Prescription in Alleviating Renal Fibrosis in UUO Mice by Regulating ERK/p38 MAPK Signaling Pathway

WANG Zhen; WANG Min; DENG Qiqi; ZHU Qing; GUO Jiao

doi:10.13422/j.cnki.syfjx.20231042

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Effect and Mechanism of Baoshen Prescription in Alleviating Renal Fibrosis in UUO Mice by Regulating ERK/p38 MAPK Signaling Pathway

更新时间：2023-10-13

- Effect and Mechanism of Baoshen Prescription in Alleviating Renal Fibrosis in UUO Mice by Regulating ERK/p38 MAPK Signaling Pathway
  增强出版
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 21, Pages: 69-78(2023)
- 作者机构：
  
  广东药科大学中医药研究院，广东省代谢病中西医结合研究中心，广东省代谢性疾病中医药防治重点实验室，广州 510006
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20231042
  CLC： R2-0;R33;R692;R289
- Published：05 November 2023，
  
  Published Online：14 August 2023，
  
  Received：04 May 2023，
- 稿件说明：
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王臻,王敏,邓琪琪等.保肾方调控ERK/p38 MAPK信号通路减轻UUO小鼠肾脏纤维化的作用及机制[J].中国实验方剂学杂志,2023,29(21):69-78.

WANG Zhen,WANG Min,DENG Qiqi,et al.Effect and Mechanism of Baoshen Prescription in Alleviating Renal Fibrosis in UUO Mice by Regulating ERK/p38 MAPK Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(21):69-78.
王臻,王敏,邓琪琪等.保肾方调控ERK/p38 MAPK信号通路减轻UUO小鼠肾脏纤维化的作用及机制[J].中国实验方剂学杂志,2023,29(21):69-78. DOI： 10.13422/j.cnki.syfjx.20231042.

WANG Zhen,WANG Min,DENG Qiqi,et al.Effect and Mechanism of Baoshen Prescription in Alleviating Renal Fibrosis in UUO Mice by Regulating ERK/p38 MAPK Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(21):69-78. DOI： 10.13422/j.cnki.syfjx.20231042.

摘要

目的

观察保肾方对肾脏纤维化的保护作用，并通过网络药理学、分子对接及体内实验探讨其可能的作用机制。

方法

将所有小鼠随机分为假手术组、模型组、保肾方低、中、高剂量组和盐酸贝那普利组。采用单侧输尿管结扎手术（UUO）制备肾脏纤维化模型，模型制备当天开始灌胃给药或蒸馏水，保肾方低、中、高剂量分别予以0.455、0.91、1.82 g·kg

-1

，盐酸贝那普利1.68 mg·kg

-1

，模型组及假手术组使用等体积蒸馏水灌胃，每天1次，连续干预14 d。检测小鼠血清尿素氮（BUN）和肌酐（Cr）水平；苏木素-伊红（HE）染色和马松（Masson）染色观察肾脏病理变化；蛋白免疫印迹法（Western blot）和免疫组化法观察肾脏纤维化相关蛋白纤维连接蛋白（FN）、

-平滑肌肌动蛋白（

-SMA）和E-钙黏蛋白（E-cadherin）的表达；实时荧光定量聚合酶链式反应（Real-time PCR）检测肾脏组织中炎症转化生长因子-

（TGF-

）、肿瘤坏死因子-

（TNF-

）、NOD样受体蛋白3（NLRP3）和单核细胞趋化蛋白-1（MCP-1）mRNA的表达；通过网络药理学、分子对接及Western blot实验探究保肾方改善肾脏纤维化的作用机制。

结果

与假手术组比较，模型组小鼠BUN、Cr含量水平显著升高（

0.01）；模型组肾小球形态结构异常，肾小管刷状缘缺失及小管扩张，蓝色胶原纤维面积明显增多；模型组小鼠FN、

-SMA含量显著升高（

0.01），E-cadherin表达显著下调（

0.01）；模型组小鼠TGF-

、TNF-

、NLRP3和MCP-1 mRNA水平表达明显增强（

0.05，

0.01）。与模型组比较，保肾方干预后，BUN、Cr含量显著降低（

0.01）；肾脏病理损害减轻，纤维化改善；FN、

-SMA表达显著下降（

0.01），E-cadherin表达显著升高（

0.01）；TGF-

、TNF-

、NLRP3、MCP-1 mRNA表达明显下降（

0.05，

0.01）。网络药理学及分子对接预测保肾方通过细胞外调节蛋白激酶（ERK）/p38丝裂原活化蛋白激酶（MAPK）信号通路发挥改善肾脏纤维化作用，药理研究表明，与假手术组比较，模型组小鼠磷酸化（p）-ERK、p-p38表达明显升高（

0.05，

0.01）；与模型组比较，保肾方中、高剂量给药处理后明显下调p-ERK、p-p38蛋白表达（

0.05，

0.01）。

结论

保肾方有效改善UUO诱导的小鼠肾脏纤维化，其作用机制可能与ERK/p38 MAPK信号通路有关。

Abstract

Objective

To observe the protective effect of Baoshen prescription against renal fibrosis and explore its underlying mechanism through network pharmacology， molecular docking， and

in vivo

experiments.

Method

All mice were randomly divided into sham surgery group， model group， low-， medium-， and high-dose Baoshen prescription groups， and a benazepril hydrochloride group. Unilateral ureteral obstruction （UUO） was performed to establish a renal fibrosis model， and the administration of Baoshen prescription at low， medium， and high doses （0.455， 0.91， and 1.82 g·kg

-1

）， and benazepril hydrochloride （1.68 mg·kg

-1

） or distilled water began on the same day as model preparation. Mice in the model group and the sham surgery group were given an equal volume of distilled water. The intervention was carried out once daily for 14 days. Mouse serum levels of blood urea nitrogen （BUN） and creatinine （Cr） were measured. Hematoxylin-eosin （HE） staining and Masson staining were used to observe renal pathological changes. Western blot and immunohistochemistry were used to assess the expression of fibronectin （FN），

-smooth muscle actin （

-SMA）， and E-cadherin， which are related to renal fibrosis. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to measure the mRNA expression of transforming growth factor-

（TGF-

）， tumor necrosis factor-

（TNF-

）， NOD-like receptor protein 3 （NLRP3）， and monocyte chemoattractant protein-1 （MCP-1） in renal tissues. The mechanism of Baoshen prescription in improving renal fibrosis was explored through network pharmacology， molecular docking， and Western blot experiments.

Result

Compared with the sham surgery group， the model group showed significantly increased levels of BUN and Cr （

0.01）. The model group exhibited abnormal renal glomerular morphology， loss of tubular brush borders， tubular dilation， and an enlarged area of blue collagen fibers. Mice in the model group showed significantly elevated levels of FN and

-SMA （

0.01）， significantly decreased expression of E-cadherin （

0.01）， and significantly increased expression of TGF-

， TNF-

， NLRP3， and MCP-1 mRNA （

0.05，

0.01）. Compared with the model group， the Baoshen prescription groups showed significantly reduced BUN and Cr levels （

0.01）， alleviated renal pathological damage， improved fibrosis， reduced expression of FN and

-SMA （

0.01）， increased E-cadherin expression （

0.01）， and downregulated mRNA expression of TGF-

， TNF-

， NLRP3， and MCP-1 （

0.05，

0.01）. Network pharmacology and molecular docking predicted that Baoshen prescription could potentially improve renal fibrosis through the extracellular signal-regulated kinase （ERK）/p38 mitogen-activated protein kinase （MAPK） signaling pathway. Pharmacological research showed that compared with the sham surgery group， the model group exhibited significantly increased expression of phosphorylated （p）-ERK and p-p38 （

0.05，

0.01）. Compared with the model group， medium- and high-dose Baoshen prescription groups showed significantly downregulated expression of p-ERK and p-p38 proteins （

0.05，

0.01）.

Conclusion

Baoshen prescription can effectively improve renal fibrosis induced by UUO in mice， and its mechanism of action may be related to the ERK/p38 MAPK signaling pathway.

关键词

肾脏纤维化中药复方单侧输尿管梗阻（UUO）网络药理学炎症丝裂原活化蛋白激酶信号通路

Keywords

renal fibrosisChinese herbal compound prescriptionunilateral ureteral obstruction （UUO）network pharmacologyinflammationmitogen-activated protein kinase（MAPK） signaling pathway

references

RUIZ-ORTEGA M，RAYEGO-MATEOS S，LAMAS S，et al.Targeting the progression of chronic kidney disease［J］.Nat Rev Nephrol，2020，16（5）：269-288.

LV J C，ZHANG L X.Prevalence and disease burden of chronic kidney disease［J］.Adv Exp Med Biol，2019，1165：3-15.

LIYANAGE T， TOYAMA T， HOCKHAM C， et al. Prevalence of chronic kidney disease in Asia：A systematic review and analysis［J］. BMJ Glob Health， 2022， 7（1）： e007525.

MACK M，YANAGITA M.Origin of myofibroblasts and cellular events triggering fibrosis［J］.Kidney Int，2015，87（2）：297-307.

LIN Y T，WU K J.Epigenetic regulation of epithelial-mesenchymal transition：Focusing on hypoxia and TGF-β signaling［J］.J Biomed Sci，2020，27（1）：39.

郭姣.治疗肾病的中药组合物、提取物、制备方法及应用和制剂：中国专利，CN202210947482.6［P］.2022-11-04.

BAI Y，WANG W，YIN P，et al.Ruxolitinib alleviates renal interstitial fibrosis in UUO mice［J］.Int J Biol Sci，2020，16（2）：194-203.

晏子友，江坚青，罗富里，等.肾衰泄浊汤及其补虚、祛邪组分对UUO大鼠肾间质纤维化的影响［J］.中国实验方剂学杂志，2020，26（16）：71-78.

WANG S，ZHOU Y，ZHANG Y，et al.Roscovitine attenuates renal interstitial fibrosis in diabetic mice through the TGF-β1/p38 MAPK pathway［J］.Biomed Pharmacother，2019，115：108895.

LIU X，TANG Z，JIANG X，et al.Cyclophilin A/CD147 signaling induces the epithelial-to-mesenchymal transition and renal fibrosis in chronic allograft dysfunction by regulating p38 MAPK signaling［J］.Ren Fail，2022，44（1）：1585-1594.

GENG X Q，MA A，HE J Z，et al.Ganoderic acid hinders renal fibrosis via suppressing the TGF-β/Smad and MAPK signaling pathways［J］.Acta Pharmacol Sin，2020，41（5）：670-677.

LEVIN A，TONELLI M，BONVENTRE J，et al.Global kidney health 2017 and beyond：A roadmap for closing gaps in care， research， and policy［J］.Lancet，2017，390（10105）：1888-1917.

WEBSTER A C，NAGLER E V，MORTON R L，et al.Chronic kidney disease［J］.Lancet，2017，389（10075）：1238-1252.

刘瑶，李伟.基于络病理论的肾间质纤维化病机及治疗初探［J］.中国中医基础医学杂志，2019，25（11）：1521-1524.

姚宁顺，王庆胜，米盼盼，等.基于“久病在络，气血皆塞”理论论治慢性肾脏病肾间质纤维化［J］.中国中医基础医学杂志，2023，29（4）：652-655.

李祥，杨文明，杨悦，等.肝豆扶木汤通过JAK/STAT信号通路对TX小鼠肾纤维化的干预作用［J］.中国实验方剂学杂志，2023，29（18）：26-35.

何珍，张孟娟，徐贺朋，等.益气活血通络方对单侧输尿管梗阻大鼠肾脏线粒体动力学的影响［J］.中国中医基础医学杂志，2023，29（4）：566-571.

蔺亚东，高文静，侯敏，等.基于网络药理学探讨芪术颗粒治疗糖尿病肾病的作用机制［J］.中国实验方剂学杂志，2020，26（24）：161-168.

杨帆，张晓云，张亚京，等. 补阳还五汤加味通过抑制炎症和纤维化改善糖尿病肾病小鼠肾脏损伤［J］. 中国实验方剂学杂志， 2022， 28（24）： 114-121.

TAN W，WANG Y，DAI H，et al.Potential therapeutic strategies for renal fibrosis：Cordyceps and related products［J］.Front Pharmacol，2022，13：932172.

ZHANG X，RITTER J K，LI N.Sphingosine-1-phosphate pathway in renal fibrosis［J］.Am J Physiol Renal Physiol，2018，315（4）：F752-F756.

HUMPHREYS B D.Mechanisms of renal fibrosis［J］. Annu Rev Physiol， 2018， 80（1）： 309-326.

CHEN L，YANG T，LU D W，et al.Central role of dysregulation of TGF-β/Smad in CKD progression and potential targets of its treatment［J］.Biomed Pharmacother，2018，101：670-681.

SUN Y B，QU X，CARUANA G，et al.The origin of renal fibroblasts/myofibroblasts and the signals that trigger fibrosis［J］.Differentiation，2016，92（3）：102-107.

HERRERA J，HENKE C A，BITTERMAN P B.Extracellular matrix as a driver of progressive fibrosis［J］.J Clin Invest，2018，128（1）：45-53.

REN J，LI J，LIU X，et al.Quercetin inhibits fibroblast activation and kidney fibrosis involving the suppression of mammalian target of rapamycin and β-catenin signaling［J］.Sci Rep，2016，6：23968.

娄原，姜雅楠，徐浩群，等.槲皮素抗脑缺血再灌注损伤作用机制研究进展［J］.中国实验方剂学杂志，2022，28（17）：248-257.

PARK Y J，BANG I J，JEONG M H，et al.Effects of β-Sitosterol from corn silk on TGF-β1-induced epithelial-mesenchymal transition in lung alveolar epithelial cells［J］.J Agric Food Chem，2019，67（35）：9789-9795.

KOC K，GEYIKOGLU F，CAKMAK O，et al.The targets of β-sitosterol as a novel therapeutic against cardio-renal complications in acute renal ischemia/reperfusion damage［J］.Naunyn Schmiedebergs Arch Pharmacol，2021，394（3）：469-479.

GUAN Y，QUAN D，CHEN K，et al.Kaempferol inhibits renal fibrosis by suppression of the sonic hedgehog signaling pathway［J］.Phytomedicine，2023，108：154246.

JIE F，YANG X，YANG B，et al.Stigmasterol attenuates inflammatory response of microglia via NF-κB and NLRP3 signaling by AMPK activation［J］.Biomed Pharmacother，2022，153：113317.

FENG S，DAI Z，LIU A B，et al.Intake of stigmasterol and β-sitosterol alters lipid metabolism and alleviates NAFLD in mice fed a high-fat western-style diet［J］.Biochim Biophys Acta Mol Cell Biol Lipids，2018，1863（10）：1274-1284.

LI R， GUO Y， ZHANG Y， et al. Salidroside ameliorates renal interstitial fibrosis by inhibiting the TLR4/NF-κB and MAPK signaling pathways［J］. Int J Mol Sci， 2019， 20（5）： 1103.

MA L，WU F，SHAO Q，et al.Baicalin alleviates oxidative stress and inflammation in diabetic nephropathy via Nrf2 and MAPK signaling pathway［J］.Drug Des Devel Ther，2021，15：3207-3221.

WU Y，WANG L，DENG D，et al.Renalase protects against renal fibrosis by inhibiting the activation of the ERK signaling pathways［J］.Int J Mol Sci，2017，18（5）： 855.

明瑞蕊，方罗昌婷，王潇潇，等.从MAPKs信号通路探讨健脾通络方的消肿镇痛作用机制［J］.中国实验方剂学杂志，2023，29（12）：85-93.

ZHANG Y，JIN D，KANG X，et al.Signaling pathways involved in diabetic renal fibrosis［J］.Front Cell Dev Biol，2021，9：696542.

LEE J，AN J N，HWANG J H，et al.p38 MAPK activity is associated with the histological degree of interstitial fibrosis in IgA nephropathy patients［J］.PLoS One，2019，14（3）：e0213981.

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