上海交通大学学报（医学版）

上海交通大学学报（医学版） >

2024 , Vol. 44 >Issue 2: 183 - 195

DOI: https://doi.org/10.3969/j.issn.1674-8115.2024.02.004

论著 · 基础研究

甲基莲心碱调节SDF-1/CXCR4信号通路对糖尿病肾病的影响

  • 王莹 ,
  • 平立风 ,
  • 刘彤彤 ,
  • 刘珊珊 ,
  • 刘磊
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  • 1.山东第一医科大学第二附属医院内分泌科，泰安 271000
    2.山东第一医科大学第二附属医院全科医学科，泰安 271000
    3.山东第一医科大学第二附属医院心血管内科，泰安 271000
    4.山东省泰安市中心医院妇科，泰安 271000
王 莹（1986—），女，副主任医师，硕士；电子信箱：taiyiwangying2009@163.com。
刘 磊，电子信箱：2625260425@qq.com。

收稿日期: 2023-04-13

  录用日期: 2023-11-30

  网络出版日期: 2024-02-28

基金资助

山东省高等学校科技计划项目(J17KA246)

收起

Effect of neferine on diabetic nephropathy by regulating SDF-1/CXCR4 signal pathway

  • Ying WANG ,
  • Lifeng PING ,
  • Tongtong LIU ,
  • Shanshan LIU ,
  • Lei LIU
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  • 1.Endocrinology Department, The Second Affiliated Hospital of Shandong First Medical University, Tai'an 271000, China
    2.Medical Category, The Second Affiliated Hospital of Shandong First Medical University, Tai'an 271000, China
    3.Cardiovascular Department, The Second Affiliated Hospital of Shandong First Medical University, Tai'an 271000, China
    4.Department of Gynaecology, Tai'an City Central Hospital of Shandong Province, Tai'an 271000, China
LIU Lei, E-mail: 2625260425@qq.com.

Received date: 2023-04-13

  Accepted date: 2023-11-30

  Online published: 2024-02-28

Supported by

Project of Science and Technology Plan of Institution of Higher Learning in Shandong Province(J17KA246)

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摘要

目的·探讨甲基莲心碱（neferine，Nef）对糖尿病肾病（diabetic nephropathy，DN）大鼠肾组织的作用及其相关机制。方法·采用高脂饲料喂食联合腹腔注射链脲佐菌素的方法构建DN模型大鼠，并将造模成功的大鼠随机分为DN组、Nef（低、中、高）剂量组、Nef高剂量+通路拮抗剂（AMD3100）组，每组10只。同时，选10只普通大鼠作为正常组。检测6组大鼠的空腹血糖（fasting blood glucose，FBG）、24 h尿蛋白、血清糖化血红蛋白（glycosylated hemoglobin，HbA1c）、血清肌酐（serum creatinine，Scr）、尿素氮（blood urea nitrogen，BUN）水平及肾指数。分别采用苏木精-伊红（hematoxylin-eosin，H-E）染色、马松（Masson）染色观察6组大鼠的肾组织的病理变化。采用硫代巴比妥酸（thiobarbituric acid，TBA）法检测肾组织丙二醛（malondialdehyde，MDA）含量，分别采用水溶性四氮唑（water soluble tetrazolium，WST-1）法、钼酸铵法检测肾组织超氧化物歧化酶（superoxide dismutase，SOD）、过氧化氢酶（catalase，CAT）的活性。分别采用实时荧光定量PCR（quantitative real-time PCR，qPCR）和蛋白质印迹法（Western blotting）检测肾组织中基质细胞衍生因子-1（stromal cell-derived factor-1，SDF-1）、CXC趋化因子受体4（CXC chemokine receptor 4，CXCR4）的mRNA以及蛋白表达。采用高糖（30 mmol/L葡萄糖）诱导大鼠肾小管上皮细胞NRK-52E，以建立DN细胞模型。将该细胞分为对照组、高糖（HG）组、HG+Nef（低、中、高）剂量组（即HG+Nef-L、M、H组）、HG+Nef-H+AMD3100组。分别采用WST-1法、钼酸铵法检测模型细胞中SOD、CAT活性，采用TBA法检测MDA含量，分别采用qPCR、Western blotting检测SDF-1、CXCR4的mRNA及蛋白表达，采用CCK-8法、流式细胞术检测细胞活力和凋亡率。结果·与DN组比较，Nef（低、中、高）剂量组和Nef高剂量+AMD3100组大鼠的FBG、24 h尿蛋白、HbA1c、Scr、BUN水平以及肾指数、MDA水平均较低，SDF-1、CXCR4的mRNA和蛋白表达以及SOD、CAT活性均较高（均P<0.05），肾组织病理损伤、纤维化程度有所减轻，且均呈剂量依赖性；AMD3100能减弱高剂量Nef对DN大鼠的肾保护作用。与HG组比较，HG+Nef-L、M、H组NRK-52E细胞的活力，SOD、CAT活性，SDF-1、CXCR4的mRNA和蛋白表达均较高，MDA含量及凋亡率均较低（均P<0.05）；AMD3100可逆转Nef-H对NRK-52E细胞损伤的保护作用。结论·Nef可能通过激活SDF-1/CXCR4信号通路来控制DN大鼠的血糖水平并提高其抗氧化能力，从而发挥肾保护作用。

关键词： 糖尿病肾病; 甲基莲心碱; 肾脏; 基质细胞衍生因子-1/CXC趋化因子受体4信号通路

本文引用格式

王莹 , 平立风 , 刘彤彤 , 刘珊珊 , 刘磊 . 甲基莲心碱调节SDF-1/CXCR4信号通路对糖尿病肾病的影响[J]. 上海交通大学学报（医学版）, 2024 , 44(2) : 183 -195 . DOI: 10.3969/j.issn.1674-8115.2024.02.004

Abstract

Objective ·To investigate the effect of neferine (Nef) on renal tissues of diabetic nephropathy (DN) rats and its related mechanism. Methods ·DN model rats were constructed by feeding high-fat diet combined with intraperitoneal injection of streptozotocin, and the successfully constructed rats were randomly divided into DN group, Nef (low, medium and high) dose groups and Nef high-dose+pathway antagonist (AMD3100) group, with 10 rats in each group. At the same time, 10 common rats were selected as the normal group. The levels of fasting blood glucose (FBG), 24 h urinary protein, serum glycosylated hemoglobin (HbA1c), serum creatinine (Scr), blood urea nitrogen (BUN) and renal index of rats in the six groups were measured. Hematoxylin-eosin (H-E) and Masson staining were used to observe the pathological changes of renal tissues. The content of malondialdehyde (MDA) in renal tissues was determined by thiobarbituric acid (TBA) method, and the activities of superoxide dismutase (SOD) and catalase (CAT) in renal tissues were determined by water soluble tetrazolium (WST-1) method and ammonium molybdate method, respectively. The mRNA and protein expressions of stromal cell-derived factor-1 (SDF-1) and CXC chemokine receptor 4 (CXCR4) in renal tissues were detected by quantitative real-time PCR (qPCR) and Western blotting, respectively. Rat renal tubular epithelium cells NRK-52E were induced by high glucose (30 mmol/L glucose) to establish DN cell model. The cells were divided into control group, high glucose (HG) group, HG+Nef (low, medium and high) dose (i.e.HG+Nef-L, M and H) group, and HG+Nef-H +AMD3100 group. SOD and CAT activities were detected by WST-1 method and ammonium molybdate method, respectively. MDA content was detected by TBA method. The mRNA and protein expressions of SDF-1 and CXCR4 were detected by qPCR and Western blotting, respectively. CCK-8 method and flow cytometry were used to detect cell viability and apoptosis rate, respecti-vely. Results ·Compared with the DN group, the levels of FBG, 24 h urinary protein, HbA1c, Scr, BUN, renal index and MDA content in Nef (low, medium and high) dose groups and Nef high-dose+AMD3100 group were decreased, the mRNA and protein expressions of SDF-1 and CXCR4 were increased, and the activities of SOD and CAT were increased (all P<0.05). The degree of pathological damage and fibrosis of renal tissues was reduced; all of the above changes were dose-dependent. AMD3100 could weaken the renal protective effect of high-dose Nef on DN rats. Compared with the HG group, NRK-52E cell viability, SOD and CAT activities, and the mRNA and protein expressions of SDF-1 and CXCR4 were increased in HG+Nef-L, M and H groups, while apoptosis rate and MDA content were decreased (all P<0.05). AMD3100 could reverse the protective effect of Nef-H on NRK-52E cell damage. Conclusion ·Nef may control blood glucose levels on DN rats and improve antioxidant capacity by activating the SDF-1/CXCR4 signal pathway, playing a renal protective role.

Key words： diabetic nephropathy (DN); neferine (Nef); renal; stromal cell-derived factor-1/CXC chemokine receptor 4 signal pathway

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