scholarly journals mTOR inhibitor INK128 promotes wound healing by regulating MDSCs

2021 ◽  
Author(s):  
Yi Li ◽  
Yujun Xu ◽  
Xinghan Liu ◽  
Xin Yan ◽  
Yue Lin ◽  
...  
Keyword(s):  
Wound Healing ◽  
High Glucose ◽  
Kinase Inhibitor ◽  
Mammalian Target Of Rapamycin ◽  
Suppressor Cells ◽  
Immunohistochemical Analysis ◽  
Mtor Signaling ◽  
Diabetic Patients ◽  
Promote Wound Healing ◽  
Healing Wound

Abstract Background: Skin wounds in diabetic patients are hardly to recover. Accumulating evidence has shown that mammalian target of rapamycin (mTOR) pathway and myeloid-derived suppressor cells (MDSCs) are involved in inflammatory-related response. INK128 is a novel mTOR kinase inhibitor in clinical development. However, the exact roles of MDSCs and INK128 in healing wound of diabetic patients are unclear. Methods: Mice models of normal, diabetic, and diabetic+INK128 were constructed. Bone marrow (BM)-derived macrophages and RAW264.7 cell line co-cultured with MDSCs, which were induced at different conditions. Flow cytometry, western blot, quantitative real-time PCR, and immunohistochemical analysis were performed. Results: Diabetic mice (DM) had a slower recovery rate, thinner epidermal and dermal, and less blood vessels than those of normal mice. MDSCs were abnormally accumulated in DM and mTOR was activated in MDSCs of DM and the cells treat with high glucose. Moreover, mTOR signaling inhibitor INK128 could promote wound healing through reducing the MDSCs. MDSCs function was disordered in DM and high glucose environments, while INK128 could help retrieve their function. Furthermore, high glucose and other factors in DM could promote M-MDSCs differentiation to M1 pro-inflammatory macrophage cells, thus inhibiting wound healing. The differentiation, which was dependent on mTOR signaling, could be reversed by INK128. Conclusion: INK128 is potential to be developed as a clinical strategy to promote wound healing of diabetic patients.

scholarly journals mTOR inhibitor INK128 promotes wound healing by regulating MDSCs

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Yi Li ◽  
Yujun Xu ◽  
Xinhan Liu ◽  
Xin Yan ◽  
Yue Lin ◽  
...  
Keyword(s):  
Wound Healing ◽  
High Glucose ◽  
Kinase Inhibitor ◽  
Mammalian Target Of Rapamycin ◽  
Suppressor Cells ◽  
Immunohistochemical Analysis ◽  
Mtor Signaling ◽  
Diabetic Patients ◽  
Promote Wound Healing ◽  
Healing Wound

Abstract Background Skin wounds in diabetic patients hardly recover. Accumulating evidence has shown that mammalian target of rapamycin (mTOR) pathway and myeloid-derived suppressor cells (MDSCs) are involved in inflammatory-related response. INK128 is a novel mTOR kinase inhibitor in clinical development. However, the exact roles of MDSCs and INK128 in healing wound of diabetic patients are unclear. Methods Mice models of normal, diabetic, and diabetic+INK128 were constructed. Bone marrow (BM)-derived macrophages and RAW264.7 cell line co-cultured with MDSCs, which were induced at different conditions. Flow cytometry, western blot, quantitative real-time PCR, and immunohistochemical analysis were performed. Results Diabetic mice (DM) had a slower recovery rate, thinner epidermis and dermis, and less blood vessels than those of normal mice. MDSCs were abnormally accumulated in DM, mTOR was activated in MDSCs of DM, and the cells were treated with high glucose. Moreover, mTOR signaling inhibitor INK128 could promote wound healing through reducing the MDSCs. MDSC function was disordered in DM and high-glucose environments, while INK128 could help retrieve their function. Furthermore, high glucose and other factors in DM could promote M-MDSC differentiation to M1 pro-inflammatory macrophage cells, thus inhibiting wound healing. The differentiation, which was dependent on mTOR signaling, could be reversed by INK128. Conclusion INK128 is potential to be developed as a clinical strategy to promote wound healing of diabetic patients.

scholarly journals mTOR Inhibitor INK128 Promotes Wound Healing by Regulating MDSCs

2020 ◽  
Author(s):  
Yi Li ◽  
Yujun Xu ◽  
Xinghan Liu ◽  
Xin Yan ◽  
Yue Lin ◽  
...  
Keyword(s):  
Wound Healing ◽  
High Glucose ◽  
Kinase Inhibitor ◽  
Mammalian Target Of Rapamycin ◽  
Suppressor Cells ◽  
Immunohistochemical Analysis ◽  
Mtor Signaling ◽  
Diabetic Patients ◽  
Promote Wound Healing ◽  
Healing Wound

Abstract Background: Skin wounds in diabetic patients are hardly to recover. Accumulating evidence has shown that mammalian target of rapamycin (mTOR) pathway and myeloid-derived suppressor cells (MDSCs) are involved in inflammatory-related response. INK128 is a novel mTOR kinase inhibitor in clinical development. However, the exact roles of MDSCs and INK128 in healing wound of diabetic patients are unclear. Methods: Mice models of normal, diabetic, and diabetic+INK128 were constructed. Bone marrow (BM)-derived macrophages and RAW264.7 cell line co-cultured with MDSCs, which were induced at different conditions. Flow cytometry, western blot, quantitative real-time PCR, and immunohistochemical analysis were performed. Results: Diabetic mice (DM) had a slower recovery rate, thinner epidermal and dermal, and less blood vessels than those of normal mice. MDSCs were abnormally accumulated in DM and mTOR was activated in MDSCs of DM and the cells treat with high glucose. Moreover, mTOR signaling inhibitor INK128 could promote wound healing through reducing the MDSCs. MDSCs function was disordered in DM and high glucose environments, while INK128 could help retrieve their function. Furthermore, high glucose and other factors in DM could promote M-MDSCs differentiation to M1 pro-inflammatory macrophage cells, thus inhibiting wound healing. The differentiation, which was dependent on mTOR signaling, could be reversed by INK128. Conclusion: INK128 is potential to be developed as a clinical strategy to promote wound healing of diabetic patients.

scholarly journals mTOR inhibitor INK128 promotes diabetic wound healing by regulating MDSCs

2020 ◽  
Author(s):  
Yi Li ◽  
Yujun Xu ◽  
Xinghan Liu ◽  
Xin Yan ◽  
Yue Lin ◽  
...  
Keyword(s):  
Wound Healing ◽  
High Glucose ◽  
Kinase Inhibitor ◽  
Mammalian Target Of Rapamycin ◽  
Suppressor Cells ◽  
Immunohistochemical Analysis ◽  
Mtor Signaling ◽  
Diabetic Patients ◽  
Healing Wound ◽  
Raw264.7 Cell

Abstract Background: Skin wounds in diabetic patients are hardly to recover. Accumulating evidence has shown that mammalian target of rapamycin (mTOR) pathway and myeloid-derived suppressor cells (MDSCs) are involved in inflammation-related response. INK128 is a novel mTOR kinase inhibitor in clinical development. However, the exact roles of MDSCs and INK128 in healing wound of diabetic patients are unclear.Methods: Mice models of normal, diabetic, and diabetic+INK128 were constructed. Bone marrow (BM)-derived macrophages and RAW264.7 cell line co-cultured with MDSCs, which were induced at different conditions. Flow cytometry, western blot, quantitative real-time PCR, and immunohistochemical analysis were performed.Results: Diabetic mice (DM) had a slower recovery rate, thinner epidermal and dermal, and less blood vessels than those of normal mice. MDSCs were abnormally accumulated in DM and mTOR was activated in MDSCs from DM or treated with high glucose. Moreover, mTOR signaling inhibitor INK128 could promoted wound healing through reducing the MDSCs. MDSCs function was disordered in DM and high glucose environments, while INK128 could help retrieve their function. Furthermore, high glucose and other factors in DM could promote M-MDSCs differentiation to M1 pro-inflammatory macrophage cells, thus inhibiting wound healing. The differentiation, which was dependent on mTOR signaling, could be reversed by INK128.Conclusion: INK128 is potential to be developed as a clinical strategy to promote wound healing of diabetic patients.

Abstract 5441: The Roles for MicroRNA-221 and IGF-1 Signaling in Regulation of High Glucose-Induced Endothelial Dysfunction and Apoptosis

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Yangxin Li ◽  
Yao-Hua Song ◽  
Fan Li ◽  
Kate McNamara ◽  
Yong-Jian Geng
Keyword(s):  
Wound Healing ◽  
Endothelial Dysfunction ◽  
High Glucose ◽  
Kinase Inhibitor ◽  
Umbilical Vein ◽  
Pi3 Kinase ◽  
High Dose ◽  
South Central ◽  
Metabolic Complication ◽  
Induced Apoptosis

Persistent hyperglycemia represents a major metabolic complication of diabetes, which may cause endothelial cell dysfunction and apoptosis. microRNAs are novel biomarkers, modulators and therapeutic targets for disease. The microRNA miR-221 is expressed in human umbilical vein endothelial cells (HUVEC). Insulin-like growth factor 1 (IGF-1) is an important survival factor for multiple cell types. This study was to investigate whether high glucose (HG) induces endothelial dysfunction and apoptosis through altering the microRNA expression and IGF-1 signaling. The mirVana qRT-PCR miRNA detection assay showed that exposure to glucose for 24 h dose-dependently (up to 25 mM) increased miR-221 expression by 2.6 ± 0.5 folds (p<0.05). HG treatment decreased c-kit protein expression (58 ± 7 % decrease vs. control at 48 h) assessed by flow cytometry. The treated HUVEC had a reduced capacity for wound healing responded to stem cell factor (SCF, the ligand of c-kit). SCF-induced HUVEC migration potential was also reduced significantly by exposure to high dose of glucose. As estimated by prediction algorithms, c-kit, but not IGF-1, was identified as the target of miR-221. Transfection of HUVEC with 2′-O-methyl antisense inhibitory oligonucleotide (AMO-221) downregulation of miR-221 led to attenuation of the glucose’s effect on the c-kit expression, SCF-induced migration and wound healing in HUVEC. Prolonged glucose exposure (25 mM, 96 h) induced apoptosis of HUVEC assessed by Death ELISA (O.D. values = 0.49 ± 0.06 in glucose group vs. 0.12 ± 0.07 in control, p<0.01). Addition of IGF-1 (30 ng/ml) blocked the glucose-induced apoptosis, whereas AMO-221 did not attenuate the apoptosis. In addition, the anti-apoptotic effects of IGF-1 was attenuated by PI3 kinase inhibitor LY294002 (15 uM) (IGF-1 + glucose + PI3 kinase inhibitor vs. IGF-1 + glucose = 0.45 ± 0.07 vs. 0.15 ± 0.05 O.D. values). Our data demonstrate that downregulation of miR-221 attenuates HG-induced reduction of c-kit, wound healing and migration of HUVEC, and enhanced IGF-1 signaling prevents HG-induced apoptosis. Thus, reducing miR-221 expression and activating IGF-1 signaling pathway may be a useful strategy to prevent diabetic-induced endothelial dysfunction and apoptosis. This research has received full or partial funding support from the American Heart Association, AHA South Central Affiliate (Arkansas, New Mexico, Oklahoma & Texas).

scholarly journals Vaccarin Regulates Diabetic Chronic Wound Healing through FOXP2/AGGF1 Pathways

2020 ◽  
Vol 21 (6) ◽  
pp. 1966
Author(s):  
Yixiao Liu ◽  
Jiangnan Sun ◽  
Xinyu Ma ◽  
Shuangshuang Li ◽  
Min Ai ◽  
...  
Keyword(s):  
Wound Healing ◽  
High Glucose ◽  
Chronic Wounds ◽  
Chronic Wound ◽  
Angiogenic Factor ◽  
Cell Counting ◽  
Sensory Loss ◽  
Diabetic Patients ◽  
And Migration ◽  
Proliferation And Migration

Background: Diabetes mellitus is a growing global health issue nearly across the world. Diabetic patients who are prone to develop diabetes-related complications often exhibit progressive neuropathy (painless and sensory loss). It is usual for small wounds to progress to ulceration, which especially worsens with peripheral arterial disease and in the presence of anaerobic bacteria, culminating into gangrene. In our study, vaccarin (VAC), the main active monomer extracted from Chinese herb vaccariae semen, is proven to have a role in promoting diabetic chronic wound healing through a cytoprotective role under high glucose conditions. Materials and methods: We constructed a pressure ulcer on both VAC-treated and control mice based on a type 1 diabetes (T1DM) model. The wound healing index was evaluated by an experimental wound assessment tool (EWAT). We also determined the effect of VAC on the proliferation and cell migration of human microvascular endothelial cells (HMEC-1) by a cell counting kit (CCK-8), a scratch and transwell assay. Results: The results demonstrated that VAC could promote the proliferation and migration of high glucose-stimulated HMEC-1 cells, which depend on the activation of FOXP2/AGGF1. Activation of the angiogenic factor with G patch and FHA domains 1 (AGGF1) caused enhanced phosphorylation of serine/threonine kinase (Akt) and extracellular regulated protein kinases (Erk1/2). By silencing the expression of forkhead box p2 (FOXP2) protein by siRNA, both mRNA and protein expression of AGGF1 were downregulated, leading to a decreased proliferation and migration of HMEC-1 cells. In addition, a diabetic chronic wound model in vivo unveiled that VAC had a positive effect on chronic wound healing, which involved the activation of the above-mentioned pathways. Conclusions: In summary, our study found that VAC promoted chronic wound healing in T1DM mice by activating the FOXP2/AGGF1 pathway, indicating that VAC may be a promising candidate for the treatment of the chronic wounds of diabetic patients.

scholarly journals Novel Application of Immunomodulatory Mushroom Polysaccharide (β-Glucan) and Triterpenes for Diabetic Wound Care

2020 ◽  
Author(s):  
Shiu-Nan Chen ◽  
Yu-Sheng Wu ◽  
Sherwin Chen ◽  
Ya-Chin Chang ◽  
Chung-Lun Lu
Keyword(s):  
Wound Healing ◽  
Ganoderma Lucidum ◽  
Cellular Proliferation ◽  
Wound Care ◽  
Immunomodulatory Activity ◽  
Diabetic Patients ◽  
Medicinal Mushroom ◽  
Positive Effects ◽  
Inflammatory Status ◽  
Healing Wound

β-Glucan and triterpenes are two important derivative compounds from traditional medicinal mushroom, such as Ganoderma lucidum and Antrodia cinnamomea. β-glucan and triterpenes are considered to have immunoregulatory properties in disease treatment for long years. The immunoregulatory effects are usually activated through some transcription of pro-inflammatory genes and possess immunomodulatory activity. Difficulty in healing wound now is a common condition that occurred in diabetic patients, and the physiological hyperglycemic status of diabetic patients resulting in the wounds continue to produce an inflammatory response. Thus, we hope to use β-glucan and triterpenes for difficult wound healing that possess immunomodulatory activity on the wound micro-environment and stimulate the positive effects on healing. In this chapter, these two important derivative compounds from traditional medicinal mushroom were examined by diabetic mammal’s wound healing models. In these models, the skin wounds’ microenvironment is expected close to diabetic foot, suffering in hyperglycemic and inflammatory status. The results are clearly presented, with the immunomodulatory effects from mushroom β-glucan and triterpenes that involved in modulating the cell-mediated immune system to cause cellular proliferation and further to introduce healing performance of the chronic inflammation wounds.

scholarly journals Verbascoside Protects Gingival Cells against High Glucose-Induced Oxidative Stress via PKC/HMGB1/RAGE/NFκB Pathway

Antioxidants ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1445
Author(s):  
Pei-Fang Hsieh ◽  
Cheng-Chia Yu ◽  
Pei-Ming Chu ◽  
Pei-Ling Hsieh
Keyword(s):  
Oxidative Stress ◽  
Wound Healing ◽  
High Glucose ◽  
Inhibitory Effect ◽  
Diabetic Patients ◽  
Impaired Wound Healing ◽  
Stress Indicator ◽  
Patients With Diabetes ◽  
Nfκb Pathway ◽  
Oral Wound Healing

Impaired wound healing often occurs in patients with diabetes and causes great inconvenience to them. Aside from the presence of prolonged inflammation, the accumulation of oxidative stress is also implicated in the delayed wound healing. In the present study, we tested the effect of verbascoside, a caffeoyl phenylethanoid glycoside, on the improvement of cell viability and wound healing capacity of gingival epithelial cells under high glucose condition. We showed that verbascoside attenuated the high glucose-induced cytotoxicity and impaired healing, which may be associated with the downregulation of oxidative stress. Our results demonstrated that verbascoside increased the activity of the antioxidant enzyme SOD and reduced the oxidative stress indicator, 8-OHdG, as well as apoptosis. Moreover, verbascoside upregulated the PGC1-α and NRF1 expression and promoted mitochondrial biogenesis, which was mediated by suppression of PKC/HMGB1/RAGE/NFκB signaling. Likewise, we showed the inhibitory effect of verbascoside on oxidative stress was via repression of PKC/HMGB1/RAGE/NFκB activation. Also, our data suggested that the PKC-mediated oxidative stress may lead to the elevated production of inflammatory cytokines, IL-6 and IL-1β. Collectively, we demonstrated that verbascoside may be beneficial to ameliorate impaired oral wound healing for diabetic patients.

Quercetin inhibits AQP1 translocation in high-glucose-cultured SRA01/04 cells through PI3K/Akt/mTOR Pathway

2020 ◽  
Vol 13 ◽  
Author(s):  
Linlin He ◽  
Nan Zhang ◽  
Lei Wang ◽  
Lei Du ◽  
Chengcheng Li ◽  
...  
Keyword(s):  
Protein Kinase ◽  
High Glucose ◽  
Protein Extraction ◽  
Protein Kinase B ◽  
Mammalian Target Of Rapamycin ◽  
Mtor Signaling ◽  
Target Of Rapamycin ◽  
Human Lens ◽  
Diabetic Cataract ◽  
Different Doses

Objective: To investigate the quercetin’s effects on Aquaporin 1 (AQP1) translocation in condition of high glucose and try to clarify the underlying mechanisms and provide new ideas for the treatment of diabetic cataract (DC). Methods: Human lens epithelial lines SRA01/04 cells were divided into groups below: normal glucose, high glucose with specific time (0 h, 2 h, 4 h, 8 h, 12 h, 24 h), high glucose plus the phosphoinositide 3-kinase (PI3K) inhibitor LY294002, high glucose plus mammalian target of rapamycin (mTOR) inhibitor rapamycin, and high glucose plus quercetin with different doses (2 μmol/L, 4 μmol/L and 8 μmol/L). Western blotting assay was used to detect the protein kinase B (Akt), phosphorylated protein kinase B (p-Akt), mammalian target of rapamycin (mTOR), phosphorylated mammalian target of rapamycin (p-mTOR) and AQP1. Real-time polymerase chain reaction (RT-PCR) was used to detect the expression of AQP1. A Membrane and Cytosol Protein Extraction Kit was applied to separate membrane proteins. Immunofluorescence assay was performed to evaluate the expression and location of AQP1. The effect of quercetin on the expression of AQP1 and PI3K/Akt/mTOR signaling was detected. Results: AQP1 protein was significantly increased at the time of 24 hour when exposured to high glucose (P<0.01). LY294002 and rapamycin inhibited PI3K/Akt/mTOR and AQP1 expression (P<0.01), prevented the change of AQP1 location in SRA01/04 plasma membrane (P<0.01). This effect was further proved by immunofluorescence. In different doses of quercetin groups (2 μmol/L, 4 μmol/L and 8 μmol/L), the phosphorylation of mTOR and Akt were decreased and AQP1 in membrane was changed compared with high glucose group (P<0.01). Conclusion: Quercetin significantly decreased the AQP1 elevation and prevented the change of AQP1 location through inhibiting the activation of the PI3K/Akt/mTOR signaling in high-glucose-cultured SRA01/04 cells, which might have the preventable and inhibitory effects on the early development of diabetic cataract. The specific pathophysiological role of quercetin still needs to be verified.

scholarly journals Hoxa3 promotes the differentiation of hematopoietic progenitor cells into proangiogenic Gr-1+CD11b+ myeloid cells

Blood ◽  
2011 ◽  
Vol 117 (3) ◽  
pp. 815-826 ◽  
Author(s):  
Elahe Mahdipour ◽  
Jayne C. Charnock ◽  
Kimberly A. Mace
Keyword(s):  
Wound Healing ◽  
Progenitor Cells ◽  
Ex Vivo ◽  
Suppressor Cells ◽  
Diabetic Patients ◽  
Differentiation Potential ◽  
Impaired Wound Healing ◽  
Immune Suppressor Cells ◽  
Bone Marrow Derived Cells

Abstract Injury induces the recruitment of bone marrow-derived cells (BMDCs) that contribute to the repair and regeneration process. The behavior of BMDCs in injured tissue has a profound effect on repair, but the regulation of BMDC behavior is poorly understood. Aberrant recruitment/retention of these cells in wounds of diabetic patients and animal models is associated with chronic inflammation and impaired healing. BMD Gr-1+CD11b+ cells function as immune suppressor cells and contribute significantly to tumor-induced neovascularization. Here we report that Gr-1+CD11b+ cells also contribute to injury-induced neovascularization, but show altered recruitment/retention kinetics in the diabetic environment. Moreover, diabetic-derived Gr-1+CD11b+ cells fail to stimulate neovascularization in vivo and have aberrant proliferative, chemotaxis, adhesion, and differentiation potential. Previously we demonstrated that gene transfer of HOXA3 to wounds of diabetic mice is taken up by and expressed by recruited BMDCs. This is associated with a suppressed inflammatory response, enhanced neovascularization, and accelerated wound healing. Here we show that sustained expression of Hoxa3 in diabetic-derived BMD Gr-1+CD11b+ cells reverses their diabetic phenotype. These findings demonstrate that manipulation of adult stem/progenitor cells ex vivo could be used as a potential therapy in patients with impaired wound healing.

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