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肾脏病与透析肾移植杂志 ›› 2022, Vol. 31 ›› Issue (6): 537-543.DOI: 10.3969/j.issn.1006-298X.2022.06.007

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鸢尾素通过上调线粒体解偶联蛋白2表达保护脓毒症相关急性肾损伤

GUO Cheng,YE Fangxiong,YANG Xueyan,LI Wei,WAN Yuhan,YANG Dingping


  

  • 出版日期:2022-12-28 发布日期:2023-01-04

Irisin plays a protective role in sepsis associated acute kidney injury by upregulating the expression of mitochondrial UCP2

  • Online:2022-12-28 Published:2023-01-04

摘要: 目的:研究鸢尾素对脓毒症相关急性肾损伤(SAAKI)是否具有保护作用及其作用机制。
方法:采用脂多糖(LPS)建立了体内和体外动物模型。用全自动生化分析仪测定血清肌酐(SCr)和血尿素氮(BUN),苏木精伊红(HE)染色观察肾脏病理改变。利用DNA 断裂的原位末端标记法(TUNEL法)检测组织细胞凋亡。免疫印迹法检测肾组织解偶联蛋白2(UCP2)、bcl2原癌基因的编码蛋白(Bcl2)、激活半胱氨酸天冬氨酸蛋白水解酶3(ccaspase3)、Bcl2样蛋白4(Bax)、白细胞介素(IL)1β、IL6、肿瘤坏死因子α(TNFα)、肾损伤分子1(KIM1)、腺苷酸活化蛋白激酶(AMPK)、磷酸化腺苷酸活化蛋白激酶(pAMPK)和甘油醛3磷酸脱氢酶(GAPDH)的表达。ELISA检测血清IL1β、IL6、TNFα的水平。RTqPCR检测UCP2、纤维连接蛋白Ⅲ型含结构域蛋白5(FNCD5)和pAMPK mRNA水平。
结果:体内实验,腹腔注射LPS导致小鼠肾功能受损、肾小管损伤评分水平升高,在补充鸢尾素后,能显著改善肾功能并降低损伤评分。研究发现鸢尾素显著降低肾组织和血清炎症因子(TNFα,IL1β和IL6)水平,改善SAAKI的炎症反应。分析Bcl2、ccaspase3、Bax的水平和TUNEL法检测结果表明,鸢尾素显著改善细胞凋亡。体外实验:测定肾近端小管上皮细胞(HK2)内ROS水平,表明鸢尾素显著改善SAAKI的氧化应激反应。通过体外实验研究鸢尾素与AMPK和UCP2的线性关系,发现鸢尾素可能是通过AMPK/UCP2信号转导途径上调SAAKI中UCP2的表达从而发挥作用。
结论:鸢尾素通过上调线粒体UCP2的表达在SAAKI中发挥保护作用。鸢尾素有望成为治疗LPS相关AKI的潜在药物。


关键词: 脓毒症, 急性肾损伤, 鸢尾素, 解偶联蛋白2, 腺苷酸活化蛋白激酶

Abstract: Objective:To investigate whether irisin has a protective effect on sepsis associated acute kidney injury (SAAKI) with sepsis and its mechanism.
Methodology:In vivo and in vitro animal models were established using LPS. Serum creatinine and blood urea nitrogen were measured by automatic biochemical analyzer. Hematoxylin eosin staining was used to observe renal pathological changes. Apoptosis was detected by TUNEL assay. The expressions of UCP2, Bcl2, ccaspase3, Bax, IL1β, IL6, TNFα, KIM1, AMPK, pAMPK and GAPDH were detected by Western blotting. The levels of IL1β, IL6 and TNFα were detected by ELISA. The mRNA levels of UCP2, FNCD5 and AMPK were detected by RTqPCR.
Results:In vivo: Intraperitoneal injection of LPS resulted in renal dysfunction and increased renal tubule injury score in mice. Irisin supplementation could significantly improve renal function and reduce the injury score. Studies showed that irisin significantly reduced the levels of inflammatory factors (TNFα, IL1β and IL6) and improved the inflammatory response of SAAKI. The levels of Bcl2, ccaspase3 and Bax and TUNEL assay showed that irisin significantly improved cell apoptosis. In vitro: ROS levels in HK2 cells were determined, indicating that irisin significantly improved the oxidative stress response of SAAKI. By studying the linear relationship between irisin and AMPK and UCP2, we found that irisin may play a role by upregulating UCP2 expression in SAAKI through AMPK/UCP2 signal transduction pathway.
Conclusion:Irisin plays a protective role in SAAKI by upregulating the expression of mitochondrial UCP2. Irisin may be a potential therapeutic agent for LPSinduced AKI.

Key words: sepsis, acute kidney injury, irisinuncoupling protein 2, AMP-activated protein kinase