Renalase is localized to the small intestine crypt and expressed upon the activation of NF-κB p65 in mice model of fasting-induced oxidative stress
Renalase is localized to the small intestine crypt and expressed upon the activation of NF-κB p65 in mice model of fasting-induced oxidative stress
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DOI：
10.1016/j.lfs.2020.118904
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发表时间：
2021-02-15
期刊：
影响因子：
6.1
通讯作者：
Takekoshi, Kazuhiro
中科院分区：
文献类型：
--
作者：
Aoki, Kai;Yanazawa, Koki;Takekoshi, Kazuhiro
目的：肾氨酸酶的表达受核因子-kappaB和低氧诱导因子-1α的调节，有关肾细胞的抗氧化应激功能已有报道。然而，肾氨酸酶的动态和在内的定位仍然未知。采用禁食诱导的氧化应激模型和Caco-2细胞模型，研究氧化应激对肾酶表达和定位的影响，并探讨其生理作用。主要方法：24只雄性小鼠随机分为对照组(CON)、禁食72 h组(Fast)和禁食后24 h组(Refeed)。分别取空肠和回肠。苏木精-伊红(HE)染色观察空肠和回肠的结构。免疫印迹法检测脑组织碳化蛋白、肾酶、核因子-kappaBp65和缺氧诱导因子-1α的表达水平。免疫荧光法观察肾素酶的定位。用Caco-2细胞进行体外实验。用腺病毒过表达肾氨酸酶。然后，用2 mM H_2O_2处理Caco-2细胞30 mm或24 h。主要结果：肾酶在Fast中升高，定位于隐窝。HIF-1α无明显升高，但核因子-kappaBp65快速升高。肾氨酸酶的过表达保护Caco-2细胞免受H_2O_2诱导的氧化应激。意义：肾氨酸酶定位于隐窝并快速增加。这种增加提示了对氧化应激的保护性反应，因为未分化的细胞定位于隐窝，需要保护。事实上，肾酶保护Caco-2细胞免受过氧化氢诱导的氧化应激。小肠肾氨酸酶的表达受核因子-kappa B p65的调节，被认为是抵抗氧化应激的一种防御机制。
Aims: Renalase expression is regulated by Nuclear Factor (NF)-kappa B and hypoxia inducible factor (HIF)-1 alpha, and antioxidative stress function in renal cells were reported. However, dynamics of renalase and localizes in in-testine were remain unknown. We evaluated the effects of oxidative stress on renalase expression and locali-zation using model of fasting induced oxidative stress and Caco-2 cell, and examined the its physiological effects.Main methods: 24 male mice were divided into three groups: Control (Con), 72 h fasting (Fast), and 24 h refeeding after fasting (Refeed). Jejunum and ileum were collected respectively. The structure of jejunum and ileum were observed by hematoxylin and eosin (HE) stain. The expression levels of carbonylated protein, renalase, NF-kappa B p65 and HIF-1 alpha were measured by immunoblotting. Localization of renalase was observed by immunofluores-cent. in vitro assay was performed using Caco-2 cell. Renalase was overexpressed using adenovirus. After that, Caco-2 cell was treated with 2 mM H2O2 for 30 mM or 24 h.Key findings: Renalase was increased in Fast and it was localized in crypt. HIF-1 alpha did not increase, but NF-kappa B p65 increased in Fast. Renalase overexpression protects the Caco-2 cells against H2O2 induced oxidative stress.Significance: Renalase was localized in crypt and increased in Fast. This increase suggested protect response to oxidative stress because undifferenced cells were localized in crypt and need to be protected. Actually, renalase protected Caco-2 cells against H2O2 induced oxidative stress. Small intestinal renalase expression was regulated by NF-kappa B p65 and was considered to be a defense mechanism against oxidative stress.