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Orientin alleviates ox-LDL-induced oxidative stress, inflammation and apoptosis in human vascular endothelial cells by regulating Sestrin 1 (SESN1)-mediated autophagy

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Abstract

Endothelial cells are a crucial component of the vessel-tissue wall and exert an important role in atherosclerosis (AS). To explore the role of Orientin in AS, human vascular endothelial cells (HUVECs) were induced by oxidized low-density lipoprotein (ox-LDL) to simulate the vascular endothelial injury during AS. Cell viability was detected by CCK-8 assay. Oxidative stress and inflammation related markers were measured using kits, RT-qPCR or western blot. Besides, cell apoptosis was assessed with TUNEL staining and cell autophagy was evaluated by LC3 immunofluorescent staining. Additionally, western blot was utilized to evaluate the expression of Sestrin 1 (SESN1) and proteins in AMPK/mTOR signaling. Afterwards, SESN1 was silenced to determine the expression of autophagy-related proteins. The further application of autophagy inhibitor 3-methyladenine (3-MA) was used to clarify the regulatory mechanism of Orientin on autophagy. Results showed that the decreased viability of HUVECs caused by ox-LDL induction was elevated by Orientin. Oxidative stress and inflammation were also attenuated after Orientin addition in HUVECs under ox-LDL condition. Moreover, Orientin suppressed apoptosis and induced autophagy of HUVECs stimulated by ox-LDL, accompanied by enhanced level of phospho (p)-AMPK and declined level of p-mTOR. Interestingly, SESN1 level was elevated by Orientin, and SESN1 depletion alleviated autophagy and reduced p-AMPK expression but enhanced p-mTOR expression. The further experiments indicated that SESN1 silencing or 3-MA addition reversed the inhibitory effects of Orientin on the oxidative stress, inflammation and apoptosis of HUVECs. Collectively, Orientin could induce autophagy by activating SESN1 expression, thereby regulating AMPK/mTOR signaling in ox-LDL-induced HUVECs.

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Data availability

The datasets generated for this study are available on request to the corresponding author.

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Authors and Affiliations

  1. Department of Cardiovascular Surgery, Xuzhou Cancer Hospital, No. 131, Huancheng Road, Xuzhou, 221005, Jiangsu, People’s Republic of China

    Feng Gao, Yongcheng Zhao, Bin Zhang, Chunwei Xiao, Zhanfa Sun, Yuan Gao & Xueyong Dou

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  1. Feng Gao
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  2. Yongcheng Zhao
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  3. Bin Zhang
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  4. Chunwei Xiao
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  5. Zhanfa Sun
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  6. Yuan Gao
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  7. Xueyong Dou
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Contributions

FG and YZ designed the study. FG, YZ, BZ, CX, ZS, YG and XD performed the experiments and analyzed the data. BZ and CX drafted the manuscript and interpreted the data. FG revised the manuscript for important intellectual content. All authors have read and approved the final version of the manuscript.

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Correspondence to Feng Gao.

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Gao, F., Zhao, Y., Zhang, B. et al. Orientin alleviates ox-LDL-induced oxidative stress, inflammation and apoptosis in human vascular endothelial cells by regulating Sestrin 1 (SESN1)-mediated autophagy. J Mol Histol 55, 109–120 (2024). https://doi.org/10.1007/s10735-023-10176-z

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