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微管相关肿瘤抑制因子1通过血红素加氧酶1抑制血红素诱导的血管内皮细胞凋亡

武生云¹, 程珂茹², 周妍芸¹, 汪引芳^1,2,*

¹上海中医药大学附属普陀医院中心实验室，上海 200062；²安徽医科大学基础医学院，合肥 230032

摘要

本研究旨在探讨微管相关肿瘤抑制因子1 (microtubule associated tumor suppressor 1, MTUS1)对血红素诱导的血管内皮细胞凋亡的影响，并探讨其对血红素加氧酶1 (hemeoxygenase 1, HMOX1)表达的影响及具体机制。采用RNA 测序、RT-qPCR和Western blot分析MTUS1敲低后人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVEC)中血红素结合相关蛋白、cAMP反应元件结合蛋白(cAMP response element-binding protein, CREB)和核呼吸因子2 (nuclear respiratory factor 2, NRF2)表达的变化，及MTUS1敲低对血红素诱导的HMOX1表达变化，CREB和NRF2过表达对MTUS1敲低介导的293T细胞中HMOX1表达变化的影响；CCK8及Western blot检测MTUS1或者HMOX1敲低对血红素诱导的HUVEC凋亡和存活的影响，NRF2过表达对MTUS1敲低介导的血红素诱导的293T细胞存活的影响。结果显示：小分子干扰RNA有效敲低HUVEC中MTUS1表达(P < 0.01)，HUVEC中MTUS1敲低后，血红素结合相关蛋白中HMOX1下调显著(P < 0.01)；MTUS1敲低能够抑制血红素诱导的HUVEC中HMOX1上调(P < 0.01)，并能促进血红素诱导的HUVEC凋亡；HMOX1敲低能促进血红素诱导的HUVEC凋亡(P < 0.01)；MTUS1敲低能抑制HUVEC中CREB和NRF2表达，NRF2过表达能部分逆转MTUS1敲低引起的293T细胞中HMOX1下调，并能部分逆转MTUS1敲低加剧的血红素诱导的293T细胞凋亡效应，但是CREB过表达不能。上述结果提示MTUS1部分通过NRF2上调HUVEC中HMOX1表达，从而抑制血红素诱导的血管内皮细胞凋亡。

关键词： 血红素; 血红素加氧酶-1; 人脐静脉内皮细胞; 凋亡; 核呼吸因子2

Microtubule-associated tumor suppressor 1 inhibits hemin-induced apoptosis of vascular endothelial cells via hemeoxygenase 1

WU Sheng-Yun¹, CHENG Ke-Ru², ZHOU Yan-Yun¹, WANG Yin-Fang^1,2,*

¹Central Laboratory, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200062, China；²School of Basic Medicine, Anhui Medical University, Hefei 230032, China

Abstract

This study aimed to investigate the effects of microtubule associated tumor suppressor 1 (MTUS1) on hemeoxygenase 1 (HMOX1) expression and hemin-induced apoptosis of vascular endothelial cells and its regulatory mechanism. RNA sequencing, RT-qPCR and Western blot were used to assess altered genes of hemin binding proteins, the expression of cAMP response element-binding protein (CREB) and nuclear respiratory factor 2 (NRF2), hemin-induced HMOX1 expression in MTUS1 knockdown human umbilical vein endothelial cells (HUVEC), and the effect of overexpression of CREB and NRF2 on HMOX1 expression in MTUS1 knockdown 293T cells. The effect of MTUS1 or HMOX1 knockdown on hemin-induced apoptosis in HUVEC, and the overexpression of NRF2 on hemin-induced apoptosis in MTUS1 knockdown 293T cells were assayed with CCK8 and Western blot. The results showed that MTUS1 was knocked down significantly in HUVEC by siRNA (P < 0.01), accompanied by decreased HMOX1 expression (P < 0.01). The increased HMOX1 expression induced by hemin was also inhibited by MTUS1 knockdown (P < 0.01). And the apoptosis of HUVEC induced by hemin was amplified by MTUS1 or HMOX1 knockdown (P < 0.01). Moreover the expression of CREB and NRF2 were both inhibited by MTUS1 knockdown in HUVEC (P < 0.01). The decreased HMOX1 regulated by MTUS1 knockdown could be rescued partly by overexpression of NRF2 (P < 0.01), however, not by overexpression of CREB. And the MTUS1 knockdown mediated decreased 293T cells viability induced by hemin could be partly rescued by NRF2 overexpression (P < 0.01). These results suggest that MTUS1 can inhibit hemin-induced apoptosis of HUVEC, and the mechanism maybe related to MTUS1/NRF2/HMOX1 pathway.

Key words: hemin; hemeoxygenase 1; human umbilical vein endothelial cell; apoptosis; nuclear respiratory factor 2

收稿日期：　　录用日期：

通讯作者：汪引芳　　E-mail: wngynfng@163.com

DOI: 10.13294/j.aps.2024.0025

引用本文：

武生云, 程珂茹, 周妍芸, 汪引芳. 微管相关肿瘤抑制因子1通过血红素加氧酶1抑制血红素诱导的血管内皮细胞凋亡[J]. 生理学报 2024; 76 (2): 215-223.

WU Sheng-Yun, CHENG Ke-Ru, ZHOU Yan-Yun, WANG Yin-Fang. Microtubule-associated tumor suppressor 1 inhibits hemin-induced apoptosis of vascular endothelial cells via hemeoxygenase 1. Acta Physiol Sin 2024; 76 (2): 215-223 (in Chinese with English abstract).