切换至 "中华医学电子期刊资源库"

中华肾病研究电子杂志 ›› 2019, Vol. 08 ›› Issue (01) : 19 -24. doi: 10.3877/cma.j.issn.2095-3216.2019.01.005

所属专题: 文献

论著

Apocynin和DPI通过抑制NOD1信号通路在缺血性AKI中的保护作用
王佳丽1, 夏田雨1, 张晓林1, 刘文丽2, 李光远2, 王军霞2, 马芳2, 覃志成1,()   
  1. 1. 030001 山西医科大学第二医院肾内科
    2. 831100 新疆五家渠人民医院肾内科
  • 收稿日期:2018-05-02 出版日期:2019-02-28
  • 通信作者: 覃志成
  • 基金资助:
    新疆兵团应用基础研究项目(2015AG017)

The protective effect of apocynin and DPI in ischemic AKI by inhibiting NOD1 signaling pathway

Jiali Wang1, Tianyu Xia1, Xiaolin Zhang1, Wenli Liu2, Guangyuan Li2, Junxia Wang2, Fang Ma2, Zhicheng Tan1,()   

  1. 1. Department of Nephrology, The Second Hospital of Shanxi Medical University, Taiyuan 030001, Shanxi Province
    2. Department of Nephrology, Wujiaqu People′s Hospital, Urumqi 831100, Xinjiang Uygur Autonomous Region; China
  • Received:2018-05-02 Published:2019-02-28
  • Corresponding author: Zhicheng Tan
  • About author:
    Corresponding author: Tan Zhicheng, Email:
引用本文:

王佳丽, 夏田雨, 张晓林, 刘文丽, 李光远, 王军霞, 马芳, 覃志成. Apocynin和DPI通过抑制NOD1信号通路在缺血性AKI中的保护作用[J/OL]. 中华肾病研究电子杂志, 2019, 08(01): 19-24.

Jiali Wang, Tianyu Xia, Xiaolin Zhang, Wenli Liu, Guangyuan Li, Junxia Wang, Fang Ma, Zhicheng Tan. The protective effect of apocynin and DPI in ischemic AKI by inhibiting NOD1 signaling pathway[J/OL]. Chinese Journal of Kidney Disease Investigation(Electronic Edition), 2019, 08(01): 19-24.

目的

NOD样受体可促发炎症反应,夹竹桃麻素(apocynin)和二苯基碘鎓(DPI)均为氧化酶抑制剂。本研究观察在缺血性急性肾损伤中抑制氧化应激产生是否能通过NOD1信号通路减轻肾间质炎症反应与细胞凋亡。

方法

将雄性Wistar大鼠随机分为4组:假手术(Sham)组、肾脏缺血再灌注(I/R)组、I/R +夹竹桃麻素(apocynin)组、I/R +二苯基碘鎓(DPI)组。通过Western印记法分别对肾组织核苷酸结合寡聚域样受体1(NOD1)、半胱天冬酶(caspase-1)及细胞核因子-κB(NF-κB)蛋白的表达进行检测;实时定量PCR法对NOD1mRNA的表达进行检测;HE染色法观察肾脏组织学改变;免疫组织化学法检测肾组织肿瘤坏死因子(TNF-ɑ)的表达;TUNEL法检测肾组织细胞凋亡。采用SPSS 22.0统计软包对实验数据进行统计学处理。

结果

与Sham组比较,I/R组大鼠肾组织NOD1、caspase-1、NF-κB、TNF-ɑ蛋白表达增加(t=16.81, t= 7.28, t= 11.08, t= 10.11;P<0.05);NOD1mRNA表达增加(t=-7.93, P<0.05);HE染色表现为急性肾小管坏死,肾小管损伤评分明显增加(t=-11.0, P<0.05);TUNEL染色显示缺血区凋亡细胞数目增加(t=-18.38, P<0.05)。与I/R组比较,I/R+ apocynin组的NOD1、caspase-1、NF-κB、TNF-ɑ蛋白表达减少(t=-10.9, t=-7.6, t=-4.9, t=-9.7;P<0.05);NOD1mRNA表达减少(t=8.49, P<0.05);HE染色后者较前者急性肾小管坏死减轻,肾小管损伤评分减低(t=-12, P<0.05);TUNEL染色显示缺血区凋亡细胞数目减少(t=-11.3, P<0.05)。与I/R组比较,I/R+DPI组的NOD1、caspase-1、NF-κB、TNF-ɑ蛋白表达减少(t=-11.4, t=-6.8, t=-5.4, t=-10.6, P<0.05);NOD1mRNA表达减少(t=7.5, P<0.05);HE染色后者较前者急性肾小管坏死减轻,肾小管损伤评分减低(t=-11, P<0.05);TUNEL染色显示缺血区凋亡细胞数目减少(t=-10.8, P<0.05)。

结论

抑制氧化应激可阻断NOD1样受体依赖的炎症途径与细胞凋亡,从而减轻肾缺血再灌注损伤。

Objective

NOD-like receptors can promote inflammation, and apocynin and diphenylene iodonium (DPI) are both oxidase inhibitors. The aim of this study was to investigate whether inhibition of oxidative stress in ischemic acute kidney injury could attenuate renal interstitial inflammatory responses and apoptosis through the NOD1 signaling pathway.

Methods

Male Wistar rats were randomly divided into 4 groups: sham operation group, renal ischemia-reperfusion (I/R) group, I/R+ apocynin group, and I/R+ DPI group. Western blotting was used to detect the renal protein expression of nucleotide-binding oligomeric domain like receptor 1 (NOD1), caspase-1, and nuclear factor-κB (NF-κB). Real-time quantitative PCR was used to detect NOD1 mRNA. The histological changes of the kidney were observed by HE staining. The expression of tumor necrosis factor-ɑ (TNF-ɑ) in renal tissue was detected by immunohistochemistry. The apoptosis of renal cells was detected by TUNEL method. Statistical analysis of experimental data was made with the SPSS 22.0 statistical soft pack.

Results

Compared with the sham group, the I/R group had higher protein expression of NOD1, caspase-1, NF-κB, and TNF-ɑ in the kidney (t=16.81, t=7.28, t=11.08, t=10.11; P<0.05), as well as higher mRNA expression of NOD1 (t=-7.93, P<0.05); HE staining showed renal lesions of acute tubular necrosis, and the renal tubular injury score increased significantly (t=-11.0, P<0.05); TUNEL staining showed that apoptotic cells increased significantly in the renal ischemia zone (t=-18.38, P<0.05). Compared with the I/R group, the I/R+ apocynin group had lower protein expression of NOD1, caspase-1, NF-κB, and TNF-ɑ in the kidney (t=-10.9, t=-7.6, t=-4.9, t=-9.7; P<0.05), as well as lower mRNA expression of NOD1 (t=8.49, P<0.05); HE staining showed lower renal lesions of acute tubular necrosis, and the renal tubular injury score reduced significantly (t=-12, P<0.05); TUNEL staining showed that apoptotic cells reduced significantly in the renal ischemia zone (t=-11.3, P<0.05). Compared with the I/R group, the I/R+ DPI group had lower protein expression of NOD1, caspase-1, NF-κB, and TNF-ɑ in the kidney (t=-11.4, t=-6.8, t=-5.4, t=-10.6, P<0.05), as well as lower mRNA expression of NOD1 (t=7.5, P<0.05); HE staining showed lower renal lesions of acute tubular necrosis, and the renal tubular injury score reduced significantly (t=-11, P<0.05); TUNEL staining showed that apoptotic cells reduced significantly in the renal ischemia zone (t=-10.8, P<0.05).

Conclusion

Inhibition of oxidative stress might block NOD1-like receptor-dependent inflammatory pathways and cellular apoptosis, thereby reducing the renal ischemia-reperfusion injury.

图1 各组大鼠肾组织NOD-1、NF-κB、Caspase-1蛋白表达(Western印迹)
图2 各组大鼠肾组织NOD1mRNA表达(实时定量PCR法)
图3 各组大鼠肾组织病理学改变(HE×200)
图4 各组大鼠肾组织TNF-ɑ表达(免疫组化×200)
图5 各组大鼠肾组织细胞凋亡比较(TUNEL染色×400)
[1]
Simonian NA,Coyle JT. Oxidative stress in neurodegenerative diseases [J]. Annual Rev Pharmacol Toxicol,2013, 36(1): 36-83.
[2]
Kleikers PW,Wingler K,Hermans JJ, et al. NADPH oxidases as a source of oxidative stress and molecular target in ischemia/reperfusion injury [J]. J Mol Med (Berl), 2012, 90(12): 1391-406.
[3]
Lambeth JD. Nox enzymes, ROS, and chronic disease: an example of antagonistic pleiotropy [J]. Free Radic Biol Med, 2007, 43(3): 332-347.
[4]
Rhoden EL,Rhoden CR,Luca ML, et al. The pathway in the renal ischemia reperfusion injury in rats [J]. Transpl Immuol, 2002, 10: 277-279.
[5]
Shigeoka AA,Kambo A,Mathison JC, et al. Nod1 and nod2 are expressed in human and murine renal tubular epithelial cells and participate in renal ischemia reperfusion injury [J]. J Immunol, 2010, 184(5): 2297-2304.
[6]
Muzaffar S,Jeremy JY,Sparatore A, et al. H2S-donating sildenafil (ACS6) inhibits superoxide formation and gp91phox expression in arterial endothelial cells: role of protein kinases A and G [J]. Br J Pharmacol, 2008, 155(7): 984-94.
[7]
Tsai PY,Ka SM,Chang JM,et al. Epigallocatechin-3-gallate prevents lupus nephritis development in mice via enhancing the Nrf2 antioxidant pathway and inhibiting NLRP3 infammasome activation [J]. Free Radic Biol Med, 2011, 51(3): 744-754.
[8]
覃志成,石媛媛,闫燕,等.硫化氢通过抑制NOD样受体途径减轻肾脏缺血再灌注损伤[J].中华肾病杂志,2014,30(8):604-608.
[9]
张日东,白瑞苗,魏敬. 盐酸小檗碱对大鼠肾小管上皮细胞缺血再灌注损伤的保护作用[J].中国实验方剂学杂志,2011,4(11):165-168.
[10]
覃志成,闫燕,李荣山.外源性硫化氢在大鼠肾脏缺血再灌注损伤中的保护作用[J]. 中华肾脏病杂志,2012,28(8):639-642.
[1] 李嘉兴, 孙乙文, 李文星. NLRP3炎性小体在急性胰腺炎中作用的研究进展[J/OL]. 中华普通外科学文献(电子版), 2024, 18(04): 300-304.
[2] 唐亦骁, 陈峻, 连正星, 胡海涛, 鲁迪, 徐骁, 卫强. 白果内酯对小鼠肝缺血再灌注损伤保护作用研究[J/OL]. 中华移植杂志(电子版), 2024, 18(05): 278-282.
[3] 王雪玲, 曹欢, 顾劲扬. 肠道菌群在器官缺血再灌注损伤中的作用及机制研究进展[J/OL]. 中华移植杂志(电子版), 2024, 18(04): 247-250.
[4] 彭瑞, 杨瑞文, 魏澹宁, 夏永良. 琥珀酸受体1加重肾脏缺血再灌注损伤的作用研究[J/OL]. 中华移植杂志(电子版), 2024, 18(03): 159-164.
[5] 张璇, 高杨, 房雅君, 姚艳玲. 保护性机械通气在肺癌胸腔镜肺段切除术中的临床应用[J/OL]. 中华肺部疾病杂志(电子版), 2024, 17(04): 563-567.
[6] 殷国青, 曾莉, 贺斌峰, 孙芬芬. Rab26负性调控Nrf2增强肺癌耐药细胞对奥希替尼的敏感性[J/OL]. 中华肺部疾病杂志(电子版), 2024, 17(03): 349-355.
[7] 孙璐, 蒋亚玲, 陈凌君. 布托啡诺对脑缺血再灌注损伤大鼠神经炎症和JAK2/STAT3信号通路的影响[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(06): 344-350.
[8] 杜霞, 马梦青, 曹长春. 造影剂诱导的急性肾损伤的发病机制及干预靶点研究进展[J/OL]. 中华肾病研究电子杂志, 2024, 13(05): 279-282.
[9] 张一绚, 韩冰, 刘超, 李思晨, 孙雪峰. 年轻化内环境改善老年小鼠肾缺血再灌注损伤诱导的肾间质纤维化[J/OL]. 中华肾病研究电子杂志, 2024, 13(03): 129-133.
[10] 李佳曦, 刘子源, 李学民. 二甲双胍对年龄相关性白内障影响的研究进展[J/OL]. 中华眼科医学杂志(电子版), 2024, 14(04): 252-256.
[11] 王子琪, 李萍, 蔡标, 杨秀敏. 雌激素在糖尿病性视网膜病变中作用机制的研究进展[J/OL]. 中华眼科医学杂志(电子版), 2024, 14(03): 187-192.
[12] 李京, 牛博, 刘晓蓓, 魏新雪, 黄荣. circ-SESN2 沉默靶向调控miRNA-23a-5p/ULK1 在神经细胞氧化应激损伤中的作用机制研究[J/OL]. 中华神经创伤外科电子杂志, 2024, 10(05): 263-272.
[13] 冯铭, 孙洪涛. 动脉瘤性蛛网膜下腔出血的颅内压监测与管理[J/OL]. 中华神经创伤外科电子杂志, 2024, 10(04): 248-253.
[14] 梁铭垚, 袁健瑜, 关雨霏, 左思程, 阳新明, 陈巧燕. 多壳扩散磁共振成像在缺血性卒中模型的应用[J/OL]. 中华介入放射学电子杂志, 2024, 12(04): 362-368.
[15] 刘俊彬, 张晓婷, 郭镜培, 刘佳妮, 于本帅, 张可, 周斌. 熊果酸通过抑制NLRP3介导的小胶质细胞焦亡减轻脑缺血再灌注损伤的研究[J/OL]. 中华介入放射学电子杂志, 2024, 12(03): 221-227.
阅读次数
全文


摘要