UBC9 介导 SUMO 化修饰在同型半胱氨酸诱导巨噬细胞焦亡中的作用研究
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1.宁夏医科大学总医院老年与特需医学科,银川 750004;2.国家卫生健康委员会代谢性心血管疾病研究重点实验室,银川 750004;3.宁夏医科大学检验学院,银川 750004;4.宁夏医科大学基础医学院,银川 750004

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R-33

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Role of UBC9-mediated SUMO modification in homocysteine-induced pyroptosis of macrophages
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1. Department of Geriatrics and Special Needs Medicine, General Hospital of Ningxia Medical University, Yinchuan 750004, China.2. Key Laboratory of Metabolic Cardiovascular Disease Research of National Health Commission, Yinchuan 750004.3. School of Laboratory Medicine, Ningxia Medical University, Yinchuan 750004. 4. Basic School of Medicine,Ningxia Medical University, Yinchuan 750004

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    摘要:

    目的 探讨泛素结合酶 9(ubiquitin conjugating enzyme 9,UBC9)介导 SUMO 化修饰在同型半胱氨酸诱导巨噬细胞焦亡中的作用。 方法 首先采用细胞计数试剂( cell counting kit-8,CCK-8)法和 Western blot 检测不同浓度(0 μmol / L、50 μmol / L、100 μmol / L、150 μmol / L 和 200 μmol / L)同型半胱氨酸对小鼠巨噬细胞(RAW264. 7)的活力及焦亡的影响;采用 Western blot 检测不同组细胞中 UBC9、SUMO 化修饰蛋白 SUMO-1、炎症因子 IL-1β 的表达水平;qRT-PCR 检测干扰前后 UBC9 的 mRNA 表达,同时检测干扰 UBC9 后,UBC9、焦亡相关蛋白、IL-1β 及SUMO-1 相关表达。 结果 当采用 100 μmol / L 的 Hcy 刺激后,巨噬细胞的活力影响最小且焦亡蛋白 NLRP3、Caspase-1 表达最明显(P<0. 001);与 Control 组相比,Hcy 组 IL-1β 的表达水平升高(P<0. 01),SUMO-1 表达升高(P<0. 01);与 Control 组相比,Hcy 组中 UBC9 蛋白水平及 mRNA 水平表达均增高(P<0. 05);转染 si-UBC9 后,与 si-NC+Hcy 组相比,si-UBC9+Hcy 组 NLRP3、Caspase-1、IL-1β、UBC9、SUMO-1 表达降低(P<0. 01)。 结论 同型半胱氨酸诱导巨噬细胞焦亡的发生,其机制与上调泛素结合酶 9 发生 SUMO 化修饰有关。

    Abstract:

    Objective To study the role of ubiquitin-conjugating enzyme 9 ( UBC9) in the pyroptosis of homocysteine-induced macrophages mediated by small ubiquitin-like modifier (SUMO) modification. Methods First, the effects of homocysteine at different concentrations (0 μmol / L, 50 μmol / L, 100 μmol / L, 150 μmol / L and 200 μmol / L) on the viability and pyrodeath of mouse macrophages (RAW264. 7) were detected by CCK-8 and Western blot. Western blot was used to detect the expression levels of UBC9, SUMO-1, and the inflammatory cytokine IL-1β in different groups of cells. qRT-PCR was used to detect the mRNA expression of UBC9 before and after RNA interference and the expression of UBC9, pyrogen-related protein, and SUMO-1 after RNA interference. Results After stimulation with 100 μmol / L homocysteine, the effect of macrophage activity was minimal, and NLRP3 and Caspase-1 were the proteins with the most obvious increase in expression (P<0. 05). Compared with the Control group, the Hcy group’ s expression of IL-1β and SUMO-1 was increased (P<0. 01). Compared with the Control group, the Hcy group’ s UBC9 protein and mRNA levels were increased (P<0. 05). The expression of NLRP3, Caspase-1, IL-1β, UBC9, and SUMO-1 was decreased in the siUBC9 + Hcy group compared with the si-NC+Hcy group (P<0. 01). Conclusions Homocysteine induces pyroptosis in macrophages, and its mechanism of action is related to the up-regulation of UBC9 to induce SUMO modification.

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马凌桔,迟宏扬,吴昕雪,马福军,田彦成,赵彩齐,何天宇,彭红建,姜怡邓,杨 力,黄 晖,马胜超. UBC9 介导 SUMO 化修饰在同型半胱氨酸诱导巨噬细胞焦亡中的作用研究[J].中国比较医学杂志,2024,34(6):11~17.

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  • 收稿日期:2024-02-27
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  • 在线发布日期: 2024-07-16
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