川芎嗪对肾结石模型大鼠肾组织氧化损伤的改善作用

doi:10.13481/j.1671-587X.20230109

摘要/Abstract

摘要：

目的探讨川芎嗪（TMP）对肾结石模型大鼠肾组织氧化损伤的改善作用，阐明其可能的作用机制。方法健康雄性SD大鼠40只，随机分为对照组、模型组、TMP组和阳性对照组，经预实验后，除对照组外其余各组大鼠采用乙醛酸盐原液80 mg·kg^－1腹腔注射构建肾结石大鼠模型，同时TMP组大鼠采用盐酸TMP注射液100 mg·kg^－1腹腔注射，阳性对照组大鼠采用肾石通颗粒3.12 g·kg^－1灌胃，对照组大鼠采用0.9%氯化钠注射液2.5 mL·kg^－1腹腔注射，连续用药10 d。取各组大鼠肾组织切片，Von Kossa染色和HE染色观察各组大鼠肾组织中钙盐沉积情况和病理形态表现，检测各组大鼠肾组织中丙二醛（MDA）水平和超氧化物歧化酶（SOD）及谷胱甘肽过氧化物酶（GSH-Px）活性。Western blotting法检测各组大鼠肾组织中核因子E2相关因子2（Nrf2）和血红素氧合酶1（HO-1）及NAD（P）H醌：氧化还原酶1（NQO1）蛋白表达水平。结果与对照组比较，模型组大鼠肾组织Von Kossa染色可见明显的钙盐沉积，结晶量化分级评分明显升高（P<0.01），HE染色可见肾组织细胞出现明显的病理损伤，肾组织中MDA水平明显升高（P<0.01），SOD和GSH-Px活性明显降低（P<0.01），肾组织中Nrf2、HO-1和NQO1蛋白表达水平明显降低（P<0.01）。与模型组比较，TMP组和阳性对照组大鼠肾组织Von Kossa染色可见钙盐沉积，结晶量化分级评分明显降低（P<0.05），HE染色可见肾组织细胞病理损伤减轻，肾组织中MDA水平明显降低（P<0.01），SOD和GSH-Px活性明显升高（P<0.05），肾组织中Nrf2、HO-1和NQO1蛋白表达水平明显升高（P<0.05）。结论肾结石可引起大鼠肾组织氧化应激损伤，导致肾组织中氧化应激指标改变和Nrf2/抗氧化反应原件（ARE）信号通路蛋白表达水平变化，TMP干预处理后可激活该信号通路，从而发挥对肾组织氧化应激损伤的改善作用。

关键词: 川芎嗪, 肾结石, 氧化应激反应, 核因子E2相关因子2, 抗氧化反应原件, 抗氧化酶

Abstract:

Objective To investigate the improvement effect of tetramethylpyrazine（TMP） on oxidative damage of kidney tissue in the nephrolithiasis model rats， and to clarify its possible mechanism. Methods A total of 40 healthy male SD rats were randomly divided into control group， model group， TMP group and positive control group. After pre-experiment，except control groups，the rats in other groups were intraperitoneally injected with 80 mg·kg^－1 glyoxylate stock solution to establish the rat models of kidney stones； the rats in TMP group were intraperitoneally injected with TMP hydrochloride injection 100 mg·kg^－1，the rats in positive control group were intragastrically perfused with 3.12 g·kg^－1 of Shenshitong granules， and the rats in control group were intraperitoneally injected with 2.5 mL·kg^－1 of 0.9% sodium chloride injection，lasted for 10 d.The rat kidney tissue sections were taken， Von Kossa staining and HE staining were performed to observe the calcium deposition and the pathomorphology of kidney tissue， and the levels of malondialdehyde（MDA） and the activities of superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px） in kidney tissue of the rats in various groups were detected. Western blotting method was used to detect the expression levels of nuclear protein nuclear factor E2-related factor 2 （Nrf2）， holoprotein heme oxygenase-1 （HO-1） and NAD（P）H quinone oxidoreductase 1 （NQO1） proteins in kidney tissue of the rats in various groups. Results Compared with control group，obvious calcium deposition was seen in the Von Kossa staining of kidney tissue of the rats in model group，and the quantitative grading score of crystal was significantly increased （P<0.01）；the HE staining results showed obvious pathological damage in kidney tissue cells，the level of MDA in kidney tissue was significantly increased （P<0.01）， the activities of SOD and GSH-Px were significantly decreased （P<0.01），and the expression levels of Nrf2， HO-1 and NQO1 proteins in kidney tissue were significantly decreased （P<0.01）. Compared with model group， calcium deposition was seen in the Von Kossa staining of kidney tissue of the rats in TMP group and positive control group， and the quantitative grading scores of crystal were decreased （P<0.05）；the HE staining results showed that the pathological damage of kidney tissue cells was alleviated， the levels of MDA were significantly decreased （P<0.05），the activities of SOD and GSH-Px were significantly increased （P<0.05）， and the expression levels of Nrf2， HO-1 and NQO1 proteins in kidney tissue were significantly increased （P<0.05）. Conclusion Kidney stones can cause oxidative stress injury in the rat kidney tissue， resulting in the changes in oxidative stress indicators and Nrf2/antioxidant response element（ARE） signaling pathway protein expression levels in kidney tissue. TMP can activate this signaling pathway after intervention， so as to improve the oxidative stress injury of kidney tissue.

Key words: Tetramethylpyrazine, Kidney stones, Oxidative stress response, Nuclear factor E2-related factor 2, Antioxidant response element, Antioxidase

中图分类号:

R692.4

程越,倪可可,傅德望,张阳,田洪阳,姜华茂. 川芎嗪对肾结石模型大鼠肾组织氧化损伤的改善作用[J]. 吉林大学学报(医学版), 2023, 49(1): 67-73.

Yue CHENG,Keke NI,Dewang FU,Yang ZHANG,Hongyang TIAN,Huamao JIANG. Improvement effect of tetramethylpyrazine on oxidative damage of kidney tissue in nephrolithiasis model rats[J]. Journal of Jilin University(Medicine Edition), 2023, 49(1): 67-73.

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