Proteomic examination of Cornus officinalis stimulated 1.1B4 human pancreatic cells reveals activation of autophagy and Keap1/Nrf2 pathway

Sharp-Tawfik, Arielle E.; Fletcher, Justin D.; Guergues, Jennifer; Marelia-Bennett, Catherine; Wolf, Tiara J.; Coiner, Alexis M.; Zhang, Y. Clare; Stevens, Stanley M.; Burkhardt, Brant R.

doi:10.1016/j.mce.2022.111773

Cited by 7 publications

(8 citation statements)

References 43 publications

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“…Nrf2 and its cytoplasmic repressor, Keap1, are master regulators under conditions of oxidative stress, which function to manage redox homeostasis, cytoprotective programming, and anti-inflammatory activities [15]. The Keap1/ Nrf2 signaling pathway has been shown to be impaired in both diabetic humans and animal models of diabetes, concurrent with increased Keap1 protein [16], suggesting a central role for modulators of oxidative stress. We previously investigated the use of synthetic ADSC-exo@MMP-PEG hydrogel in wound treatment, demonstrating that it could alleviate H 2 O 2 -induced oxidative stress and rescue the impaired wound healing in diabetic mice [17].…”

Section: Discussionmentioning

confidence: 99%

Milk-derived exosomes carrying siRNA-KEAP1 promote diabetic wound healing by improving oxidative stress

Xiang

Chen

Jiang

et al. 2023

Drug Deliv. and Transl. Res.

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Diabetic wounds are a serious complication of diabetes mellitus (DM) that can lead to persistent infection, amputation, and even death. Prolonged oxidative stress has been widely recognized as a major instigator in the development of diabetic wounds; therefore, oxidative stress is considered a promising therapeutic target. In the present study, Keap1/Nrf2 signaling was confirmed to be activated in streptozotocin (STZ)-induced diabetic mice and methylglyoxal (MGO)-treated human umbilical vein endothelial cells (HUVECs). Knockdown of Keap1 by siRNA reversed the increase in Keap1 levels, promoted the nuclear translocation of Nrf2, and increased the expression of HO-1, an antioxidant protein. To explore therapeutic delivery strategies, milk-derived exosomes (mEXOs) were developed as a novel, efficient, and non-toxic siRNA carrier. SiRNA-Keap1 (siKeap1) was loaded into mEXOs by sonication, and the obtained mEXOs-siKeap1 were found to promote HUVEC proliferation and migration while relieving oxidative stress in MGO-treated HUVECs. Meanwhile, in a mouse model of diabetic wounds, injection of mEXOs-siKeap1 significantly accelerated diabetic wound healing with enhanced collagen formation and neovascularization. Taken together, these data support the development of Keap1 knockdown as a potential therapeutic strategy for diabetic wounds and demonstrated the feasibility of mEXOs as a scalable, biocompatible, and cost-effective siRNA delivery system. Graphical Abstract The therapeutic effect of siKeap1-loaded mEXOs on diabetic wound healing was assessed. First, we found that the expression of Keap1 was upregulated in the wounds of diabetic mice and in human umbilical vein endothelial cells (HUVECs) pretreated with methylglyoxal (MGO). Next, we extracted exosomes from raw milk by differential centrifugation and loaded siKeap1 into milk-derived exosomes by sonication. The in vitro application of the synthetic complex (mEXOs-siKeap1) was found to increase the nuclear localization of Nrf2 and the expression of the antioxidant protein HO-1, thus reversing oxidative stress. Furthermore, in vivo mEXOs-siKeap1 administration significantly accelerated the healing rate of diabetic wounds (Scheme 1). Scheme 1 Schematic diagram. A Synthesis of mEXOs-siKeap1 complex. B Mechanism of mEXOs-siKeap1 in vitro. C The treatment effect of mEXOs-siKeap1 on an in vivo mouse model of diabetic wounds Supplementary Information The online version contains supplementary material available at 10.1007/s13346-023-01306-x.

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Section: Discussionmentioning

confidence: 99%

Milk-derived exosomes carrying siRNA-KEAP1 promote diabetic wound healing by improving oxidative stress

Xiang

Chen

Jiang

et al. 2023

Drug Deliv. and Transl. Res.

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“…Nrf2, a transcription factor, and its cytoplasmic repressor Keap1 form the master regulator when cells are under oxidative stress, which could manage redox homeostasis, cytoprotective programming and anti-in ammatory activities [13]. It was veri ed that the Keap1/Nrf2 signaling pathway was impaired and the level of Keap1 protein was down-regulated, both in diabetic human and animal models [14]. In wound healing treatment, our research team had researched that the synthetic ADSC-exo@MMP-PEG hydrogel could alleviate the H 2 O 2 -induced oxidative stress and improve the impaired wound healing in diabetes mice [15] and the thermosensitive niobium carbide (Nb 2 C)-based hydrogel (Nb 2 C@Gel) with antioxidative and antimicrobial activity could promote wound healing by attenuating ROS levels [16].…”

Section: Discussionmentioning

confidence: 99%

Milk-derived exosomes carrying siRNA-KEAP1 promote diabetic wound healing by improving oxidative stress

Xiang

Chen

Jiang

et al. 2022

Preprint

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Diabetic wound, as a serious complication of diabetes mellitus (DM), leads to persistent infection, amputation and even death. Prolonged oxidative stress has been widely recognized as the main cause of diabetic wound and is considered as hopeful therapeutic target. In the present study, Keap1/Nrf2 signaling was confirmed to be activated in streptozotocin (STZ)-induced diabetic mice and methylglyoxal (MGO)-treated human umbilical vein endothelial cells (HUVEC). Knockdown of Keap1 by siRNA attenuated the raised Keap1 level, promoted the nuclear transfer of Nrf2, and then facilitated the expression of HO-1, an antioxidant protein. Then the milk-derived exosomes (mEXOs) were developed as a novel, efficient and non-toxic siRNA carrier. SiRNA-Keap1 (siKeap1) was loaded into mEXOs through ultrasonic technique, which promoted HUVEC proliferation, migration and relieved oxidative stress in MGO-treated HUVEC. In the meantime, the injection of mEXOs-siKeap1 significantly accelerated the diabetic wound healing with enhanced collagen formation and neovascularization in mice models. Totally, the present study puts forward a potential treatment for diabetic wound and proved the feasibility of mEXOs as a scalable, biocompatible, and cost-effective siRNA delivery system.

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“…Peripheral insulin sensitivity is attenuated by increased adiposity that elevates free fatty acid levels in the circulation. Acute exposure of β-cells to elevated glucose levels causes reversible insensitivity to glucose whereas chronic exposure may either cause irreversible loss of β-cell function and apoptosis (glucotoxicity) [3,4].…”

Section: Introductionmentioning

confidence: 99%

Proteomic profiling of exosomes derived from pancreatic beta-cells cultured under hyperglycemia

Rząca

Jankowska

Stępień

2022

Bio-Algorithms and Med-Systems

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Introduction Cargo carried by extracellular vesicles (EVs) is considered a promising diagnostic marker, especially proteins. EVs can be divided according to their size and way of biogenesis into exosomes (diameter < 200 nm) and ectosomes (diameter > 200 nm). Exosomes are considered to be of endocytic origin, and ectosomes are produced by budding and shedding from the plasma membrane [1]. Methods The first step of this study was a characterization of the exosome sample. Using Tunable Resistive Pulse Sensing (qNano) size distribution and concentration were measured. The mean size of exosomes was 120±9.17 nm. In the present study, a nano liquid chromatography coupled with tandem mass spectrometry (nanoLCMS/MS) was used to compare protein profiles of exosomes secreted by pancreatic beta cells (1.1B4) grown under normal glucose (NG, 5 mM D-glucose) and high glucose (HG, 25 mM D-glucose) conditions. The EV samples were lysed, and proteins were denatured, digested, and analyzed using a Q-Exactive mass spectrometer coupled with the UltiMate 3000 RSLC nano system. The nanoLC-MS/MS data were searched against the SwissProt Homo sapiens database using MaxQuant software and protein quantitation was done by the MaxLFQ algorithm. Statistical analysis was carried out with Perseus software. Further bioinformatic analysis was performed using the FunRich 3.1.4 software with the UniProt protein database and String [2]. Results As a result of the nanoLC-MS/MS analysis more than 1,000 proteins were identified and quantified in each sample. The average number of identified proteins in exosomes was 1,397. Label-free quantitative analysis showed that exosome composition differed significantly between those isolated under NG and HG conditions. Many pathways were down-regulated in HG, particularly the ubiquitin-proteasome pathway. In addition, a significant up-regulation of the Ras-proteins pathway was observed in HG. Conclusion Our description of exosomes protein content and its related functions provides the first insight into the EV interactome and its role in glucose intolerance development and diabetic complications. The results also indicate the applicability of EV proteins for further investigation regarding their potential as circulating in vivo biomarkers.

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Proteomic examination of Cornus officinalis stimulated 1.1B4 human pancreatic cells reveals activation of autophagy and Keap1/Nrf2 pathway

Cited by 7 publications

References 43 publications

Milk-derived exosomes carrying siRNA-KEAP1 promote diabetic wound healing by improving oxidative stress

Milk-derived exosomes carrying siRNA-KEAP1 promote diabetic wound healing by improving oxidative stress

Milk-derived exosomes carrying siRNA-KEAP1 promote diabetic wound healing by improving oxidative stress

Proteomic profiling of exosomes derived from pancreatic beta-cells cultured under hyperglycemia

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