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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

29卷

期数:

2023年02期

页码:

153-160

栏目:

基础医学

出版日期:

2023-03-20

文章信息/Info

Title:

Mechanism of FushenJiangzhuo recipe and lumbrokinase on renal interstitial fibrosis based on RNA-seq

文章编号:

1006-8147（2023）02-0153-08

作者:

王超坤; 刘婷婷; 帅一尘; 李春雨; 苏玮莲; 宁志芬; 李国霞

天津医科大学基础医学院药 理学系，天津300070

Author(s):

WANG Chao-kun; LIU Ting-ting; SHUAI Yi-chen; LI Chun-yu; SU Wei-lian; NING Zhi-fen; LI Guo-xia

Department of Pmarmacology，School of Basic Medical Sciences，Tianjin Medical University，Tianjin 300070，China

关键词:

扶肾降浊方; 蚓激酶; 肾间质纤维化; Flna; ERK

Keywords:

FushenJiangzhuo recipe; lumbrokinase; renal interstitial fibrosis; Flna; ERK

分类号:

R692.9

DOI:

-

文献标志码:

A

摘要:

目的：探讨扶肾降浊方、蚓激酶及联合用药对肾间质纤维化的作用机制。方法：雄性Sprauge-Dawley（SD）大鼠60只，随机分为假手术组、模型组、扶肾降浊方组、蚓激酶组和联合用药组，每组12只，术后第2天开始，每天给药1次，扶肾降浊方灌胃给药：29g/（kg·d），蚓激酶腹腔注射给药：36万U/（kg·d），假手术组、模型组给予等体积生理盐水，于第7、14天剖杀。HE和Masson染色评价纤维化改变；RNA测序（RNA-seq）检测第7天模型组和治疗组的基因表达；Western印迹检测第7、14天肾脏中纤维化标志蛋白[纤溶酶原激活物抑制剂-1（PAI-1）、Collagen-1、纤连蛋白（Fibronectin）]、上皮-间充质转化（EMT）标志蛋白（E-cad、-SMA、Vimentin）、Flna、磷酸化细胞外信号调节激酶（p-ERK）和细胞外信号调节激酶（ERK）的蛋白水平。结果：病理显示造模7天后肾脏出现明显纤维化改变（P<0.01），随造模时延长而加重（P<0.01）；扶肾降浊方、蚓激酶及联合用药对纤维化有明显抑制作用（P<0.05或P<0.01），第7天，扶肾降浊方较蚓激酶作用更为明显（P<0.01）；第14天，蚓激酶较扶肾降浊方作用更为明显（P<0.05），两个时期均以联合用药效果最佳。同时，造模后肾脏中的纤维化和EMT标志蛋白显著升高（均P<0.01），扶肾降浊方、蚓激酶及联合用药对纤维化和EMT标志蛋白均有明显抑制作用（P<0.05或P<0.01），第7天，扶肾降浊方较蚓激酶作用更为明显（P<0.05或P<0.01）；第14天，蚓激酶较扶肾降浊方作用更为明显（P<0.05或P<0.01），两个时期均以联合用药作用最明显（P<0.05或P<0.01）。机制上，第7、14天，造模后各组Flna和p-ERK蛋白表达均明显升高（均P<0.01），扶肾降浊方、蚓激酶及联合用药均可抑制Flna和p-ERK的表达（P<0.05或P<0.01）。结论：扶肾降浊方、蚓激酶及联合用药均能抑制肾间质纤维化，病变早期应用扶肾降浊方效果优于蚓激酶且联合用药优于单独用药，其共同机制可能通过抑制Flna基因及ERK的磷酸化，抑制表型转化，减少细胞外基质的产生而起作用。

Abstract:

Objective：To investigate the mechanisms of FushenJiangzhuo Recipe（FSJZ），lumbrokinase and their combination on renal interstitial fibrosis（RF）. Methods：Sixty male Sprauge-Dawley（SD）rats were randomly divided into sham operation group，model group， FSJZ group，lumbrokinase group and combined group，with 12 rats in each group. Starting from the second day after surgery，the drug was given once a day. The FSJZ was given intragastric administration：29 g/（kg·d），and the lumbrokinase intrabitoneal injection：360 000 U/（kg·d）. The sham group and model group were given the same volume of normal saline and dissected on the 7th and 14th day. HE and Masson staining were used to evaluating the fibrosis changes. RNA sequencing（RNA-seq）was used to detect gene expression in the model group and treatment group on day 7. Western blotting was used to detect the protein levels of RF marker proteins [plasminogen activator inhibitor-1（PAI-1），Collagen-1，fibronectin]，epithelial-mesenchymal transition（EMT）marker proteins（E-cad，α-SMA，Vimentin）， Flna，phosphorylated extracellular signal-regulated kinase（p-ERK）and extracellular signal-regulated kinase（ERK）in the kidney on the 7，14 days. Results：Pathological findings showed obvious RF changes after modeling（P<0.01）and increased with the lengthening of mold-making time（P<0.01）.FSJZ，lumbrokinase and their combination could significantly inhibit fibrosis（P<0.05 or P<0.01）on the 7th day，the effect of FSJZ was more obvious than that of lumbrokinase（P<0.01）.On day 14，the effect of lumbrokinase was more obvious than that of FSJZ （P<0.05），the best effect was found in combination therapy in both periods. At the same time，RF and EMT marker proteins were significantly increased after modeling（both P<0.01），FSJZ，lumbrokinase and their combination had significant inhibitory effects on fibrosis and EMT marker protein（P<0.05 or P<0.01），on the 7th day，the effect of FSJZ was more obvious than that of lumbrokinase （P<0.05 or P<0.01）；On day 14，the effect of lumbrokinase was more obvious than that of FSJZ（P<0.05 or P<0.01），the most obvious effect was found in combination therapy（P<0.05 or P<0.01）.In terms of mechanism，on the 7th and 14th day，the expression of Flna and p-ERK protein increased significantly in each group after modeling（both P<0.01），FSJZ，lumbrokinase and their combination could inhibit the expression of Flna and p-ERK（P<0.05 or P<0.01）. Conclusion：FSJZ，lumbrokinase and combination can inhibit RF. The effect of FSJZ is better than lumbrokinase at the early stage of lesions，and the combination is better than single drug. The common mechanism may be through inhibiting Flna and ERK phosphorylation，inhibiting EMT，and reducing the production of extracellular matrix.

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相似文献/References:

[1]王晓敏,申珅,刘婷婷,等.蚓激酶对UUO大鼠肾组织NOX4、FAK、Src的影响[J].天津医科大学学报,2021,27(03):234.
　WANG Xiao-min,SHEN Shen,LIU Ting-ting,et al.Effects of lumbrokinase on expression of NOX4, FAK, Src in renal interstitial fibrosis of unilateral ureteral obstruction（UUO）rats[J].Journal of Tianjin Medical University,2021,27(02):234.

备注/Memo

备注/Memo:

基金项目： 天津市卫生健康委员会中医中西医结合科研项目（2021032）
作者简介： 王超坤（1997-），女，硕士在读，研究方向：中西医结合治疗肾纤维化；
通信作者：李国霞，E-mail：liguoxia96@163.com。

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更新日期/Last Update: 2023-04-30