青蒿琥酯通过NLRP3炎症小体抑制细胞焦亡减轻大鼠肾缺血-再灌注损伤

袁强; 申开文; 张瑞波; 王强; 吴玉婷; 沈俊

doi:10.3969/j.issn.1674-7445.2021.06.014

青蒿琥酯通过NLRP3炎症小体抑制细胞焦亡减轻大鼠肾缺血-再灌注损伤

doi: 10.3969/j.issn.1674-7445.2021.06.014

550004 贵阳，贵州医科大学附属医院泌尿外科（袁强、申开文、张瑞波、王强、沈俊），病理科（吴玉婷）

基金项目:

贵州省卫生健康委科学技术基金 gzwkj2021-220

贵州省科技计划项目（2018）5779-6

国家自然科学基金培育项目（贵州医科大学附属医院） gyfynsfc（2020）-30

详细信息

作者简介:
袁强，男，1993年生，硕士研究生，研究方向为肾移植相关缺血-再灌注损伤与保护，Email: 411057131@qq.com

通讯作者:
沈俊，男，1973年生，博士，副主任医师，研究方向为肾移植相关缺血-再灌注损伤与保护，Email: shenjun@gmc.edu.cn

中图分类号: R617
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出版历程
- 收稿日期: 2021-07-22
- 刊出日期: 2021-11-15

Artesunate alleviates renal ischemia-reperfusion injury in rats by inhibiting pyroptosis via NLRP3 inflammasome

Department of Urology Surgery, the Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China

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Corresponding author: Shen Jun, Email: shenjun@gmc.edu.cn

摘要

摘要: 目的探究青蒿琥酯对大鼠肾缺血-再灌注损伤（IRI）的作用及机制。方法将25只SD大鼠随机分为假手术组（Sham组）、模型组（IRI组）、低剂量青蒿琥组（ART-L组）、高剂量青蒿琥酯组（ART-H组）和NLRP3炎症小体抑制剂组（INF39组），每组5只。分析各组大鼠血清肌酐（Scr）、血尿素氮（BUN）水平及肾组织病理损伤情况；检测各组大鼠血清肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6水平；用免疫组织化学染色法检测各组大鼠肾组织IL-1β的表达；免疫荧光染色和蛋白质免疫印迹法检测焦亡相关蛋白的表达。结果与Sham组比较，IRI组Scr和BUN水平升高，肾组织损伤较重，炎症因子TNF-α、IL-6、IL-1β表达水平升高，肾损伤分子（KIM）-1及焦亡相关蛋白NOD样受体蛋白3（NLRP3）、半胱氨酸天冬氨酸蛋白酶（Caspase）-1、Gasdermin D（GSDMD）、IL-1β蛋白表达水平均升高；与IRI组比较，ART-L组、ART-H组和INF39组Scr和BUN水平均下降，肾组织损伤均较轻，TNF-α、IL-6、IL-1β表达水平均降低，KIM-1、NLRP3、Caspase-1、GSDMD、IL-1β蛋白表达水平均下降。结论青蒿琥酯可抑制NLRP3炎症小体诱导的细胞焦亡，减少肾IRI后焦亡相关蛋白的表达及炎症因子的释放，减轻肾IRI。
- 青蒿琥酯 /
- 肾移植 /
- 缺血-再灌注损伤 /
- NOD样受体蛋白3 /
- 细胞焦亡 /
- 炎症 /
- 血清肌酐 /
- 血尿素氮
Abstract: Objective To investigate the effect and mechanism of artesunate on renal ischemia-reperfusion injury (IRI) in rat. Methods Twenty-five SD rats were randomly divided into the sham operation group (Sham group), model group (IRI group), low-dose artesunate group (ART-L group), high-dose artesunate group (ART-H group) and NLRP3 inflammasome inhibitor group (INF39 group), with 5 rats in each group. The levels of serum creatinine (Scr), blood urea nitrogen (BUN) and pathological damage of renal tissue were analyzed. The levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in serum were quantitatively measured. The expression level of IL-1β in renal tissues was determined by immunohistochemical staining. The expression levels of pyroptosis-related proteins were detected by fluorescent staining and Western blot. Results Compared with the Sham group, the levels of Scr and BUN were higher, the renal tissue injury was aggravated, the expression levels of TNF-α, IL-6 and IL-1βwere higher, and the expression levels of kidney injury molecule (KIM-1), pyroptosis-related protein NOD-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase (Caspase-1), Gasdermin D (GSDMD) and IL-1β proteins were higher in the IRI group. Compared with the IRI group, the levels of Scr and BUN were decreased, the renal tissue injury was mitigated, the expression levels of TNF-α, IL-6 and IL-1β were down-regulated, and the expression levels of KIM-1, NLRP3, Caspase-1, GSDMD and IL-1β proteins were down-regulated in the ART-L, ART-H and INF39 groups. Conclusions Artesunate may inhibit pyroptosis induced by NLRP3 inflammasome, down-regulate the expression levels of pyroptosis -related proteins, reduce the release of inflammatory factors after renal IRI and alleviate renal IRI.
- Artesunate /
- Kidney transplantation /
- Ischemia-reperfusion injury /
- NOD-like receptor protein 3 /
- Pyroptosis /
- Inflammation /
- Serum creatinine /
- Blood urea nitrogen

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图 1 各组大鼠肾组织病理学表现

注: A图为各组大鼠肾IRI后24 h肾组织病理学结果(HE，x400); B图为各组大鼠肾IRI后24 h肾组织病理学结果(PAS，x200)。

Figure 1. Renal histopathological features of rats in each group

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图 2 各组大鼠肾功能血清学指标

注: A图为各组大鼠肾IRI后24hBUN水平; B图为各组大鼠肾IRI后24hScr水平; 与Sham组比较，^aP<0.05;与IRI组比较，^bP<0.05;与ART-L组比较，^cP<0.05。

Figure 2. Serum indexes of renal function of rats in each group

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图 3 各组大鼠肾组织KIM-1蛋白表达水平

注: 与Sham组比较，^aP<0.05; 与IRI组比较，^bP<0.05; 与ART-L组比较，^cP<0.05; 与ART-H组比较，^dp<0.05。

Figure 3. The expression levels of KIM-1 protein in renal tissues of rats in each group

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图 4 各组大鼠肾组织IL-1β的表达情况

注: 图示各组大鼠肾IRI后24 h肾组织IL-β的表达(免疫组化，x 200)

Figure 4. Expression of IL-1 in renal tissues of rats in each group

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图 5 各组大鼠血清炎症因子的水平

注: A图示各组大鼠肾IRI后24 h血清TNF-ax水平; B图示各组大鼠肾IRI后24 h血清IL-6水平; 与Sham组比较，^aP<0.05; 与IRI组比较，^bP<0.05; 与ART-L组比较，^cP<0.05。

Figure 5. The levels of inflammatory factors in serum of rats in each group

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图 6 各组大鼠肾组织焦亡相关蛋白的表达情况

注: A图为各组大鼠肾组织焦亡相关蛋白表达情况，与Sham组比较，^aP<0.05, 与IRI组比较，^bP<0.05，与ART-L组比较，^cP<0.05;B图为各组大鼠肾IRI后24 h肾组织NLRP3. Caspase-1的表达情况(免疫荧光，x200); C图为各组大鼠肾IRI后24 h肾组织GSDMD的表达情况(免疫荧光，x200)。

Figure 6. Expression of pyroptosis related proteins in renal tissues of rats in each group

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