Background:Wound management of diabetic foot ulcers(DFUs)is a complex and challenging task,and existing strategies fail to meet clinical needs.Therefore,it is important to develop novel drug candidates and discover ne...Background:Wound management of diabetic foot ulcers(DFUs)is a complex and challenging task,and existing strategies fail to meet clinical needs.Therefore,it is important to develop novel drug candidates and discover new therapeutic targets.However,reports on peptides as molecular probes for resolving issues related to DFUs remain rare.This study utilized peptide RL-QN15 as an exogenous molecular probe to investigate the underlying mechanism of endogenous non-coding RNA in DFU wound healing.The aim was to generate novel insights for the clinical management of DFUs and identify potential drug targets.Methods:We investigated the wound-healing efficiency of peptide RL-QN15 under diabetic con-ditions using in vitro and in vivo experimental models.RNA sequencing,in vitro transfection,quantitative real-time polymerase chain reaction,western blotting,dual luciferase reporter gene detection,in vitro cell scratches,and cell proliferation and migration assays were performed to explore the potential mechanism underlying the promoting effects of RL-QN15 on DFU repair.Results:Peptide RL-QN15 enhanced the migration and proliferation of human immortalized keratinocytes(HaCaT cells)in a high-glucose environment and accelerated wound healing in a DFU rat model.Based on results from RNA sequencing,we defined a new microRNA(miR-4482-3p)related to the promotion of wound healing.The bioactivity of miR-4482-3p was verified by inhibiting and overexpressing miR-4482-3p.Inhibition of miR-4482-3p enhanced the migration and proliferation ability of HaCaT cells as well as the expression of vascular endothelial growth factor B(VEGFB).RLQN15 also promoted the migration and proliferation ability of HaCaT cells,and VEGFB expression was mediated via inhibition of miR-4482-3p expression by the p38 mitogen-activated protein kinase(p38MAPK)and smad3 signaling pathways.Conclusions:RL-QN15 is an effective molecule for the treatment of DFUs,with the underlying mechanism related to the inhibition of miR-4482-3p expression via the p38MAPK and smad3 signaling pathways,ultimately promoting re-epithelialization,angiogenesis and wound healing.This study provides a theoretical basis for the clinical application of RL-QN15 as a molecular probe in promoting DFU wound healing.展开更多
In order to investigate the promoting effect of low-intensity treadmill exercise on rat dorsal wound healing and the mechanism, 20 Sprague-Dawley rats were randomly divided into two groups： exercise group（Ex） and n...In order to investigate the promoting effect of low-intensity treadmill exercise on rat dorsal wound healing and the mechanism, 20 Sprague-Dawley rats were randomly divided into two groups： exercise group（Ex） and non-exercise group（non-ex）. The rats in Ex group were given treadmill exercise for one month, and those in non-ex group raised on the same conditions without treadmill exercise. Both groups received dorsal wound operation with free access to food and water. By two-week continuous observation and recording of the wound area, the healing rate was analyzed. The blood sample was collected at day 14 post-operation via cardiac puncture for determination of the number of endothelial progenitor cells（EPCs） by flow cytometry, and the concentrations of relevant cytokines such as basic fibroblast growth factor（b FGF）, endothelial nitric oxide synthase（e NOS） and vascular endothelial growth factor（VEGF） were measured by ELISA. The skin tissue around the wound was dissected to observe the vascular density under the microscope after HE staining, to detect the m RNA level of VEGFR2 and angiopoietin-1（Ang-1） receptor using RT-q PCR, and protein expression of α-smooth muscle actin（αSMA） and type collagenⅢ（ColⅢ） using Western blotting. It was found that the wound area in Ex group was smaller at the same time point than in non-ex group. The number of circulating EPCs was greater and the concentrations of vasoactive factors such as VEGF, e NOS and b FGF were higher in Ex group than in non-ex group. HE staining displayed a higher vessel density in Ex group than in non-ex group. Moreover, the m RNA expression of VEGFR2 and Ang-1 detected in the wound tissue in Ex group was higher than in non-ex group. Meanwhile, the protein expression of αSMA and Col Ⅲwas more abundant in Ex group than in non-ex group. Conclusively, the above results demonstrate Ex rats had a higher wound healing rate, suggesting low-intensity treadmill exercise accelerates wound healing. The present work may provide some hint for future study of treating refractory wound.展开更多
Objective:Elet-rich plasma(PRP)combined with adipose-derived mesenchymal stem cells(ADSC)on skin wound healing and serum hyaluronic acid(HA)and vascular endothelial growth factor(VEGF)levels in mice.Methods:Forty-five...Objective:Elet-rich plasma(PRP)combined with adipose-derived mesenchymal stem cells(ADSC)on skin wound healing and serum hyaluronic acid(HA)and vascular endothelial growth factor(VEGF)levels in mice.Methods:Forty-five mice were randomly divided into three groups(n=15),which were control,ADSC,and ADSC+PRP.The wound injury model was established in all three groups.The control group was injected subcutaneously with normal saline,the ADSC group was injected subcutaneously with allogeneic PRP,and the ADSC+PRP was injected subcutaneously with ADSC and PRP.Wound healing was observed at 3 d,5 d,and 7 d after modeling,and histomorphological features were observed by HE staining.CollagenⅠand CollagenⅢlevels were detected using Western blot.The levels of serum HA and VEGF in each group were detected and compared.Results:The wound healing rate of ADSC group and ADSC+PRP group was significantly higher than that of control group at 3 d,5 d and 7 d after modeling(P<0.01).The wound healing rate of the ADSC+PRP group was significantly higher than that of the ADSC group(P<0.01).The inflammatory response of the control group and the ADSC group was significant at 3 and 5 d after modeling,and there was a small amount of granulation tissue in the ADSC group.The ADSC+PRP group had a milder inflammatory response and more granulation tissue;After 7 d,the ADSC+PRP group had more neovascularization and higher re-epithelialization rate.The control group and the ADSC group had poorer epithelialization and fewer new blood vessels.The expression levels of CollagenⅠand CollagenⅢin ADSC group and ADSC+PRP group were significantly higher than those in control group at 3 d,5 d and 7 d after modeling(P<0.01).CollagenⅠand CollagenⅢin the ADSC+PRP group were significantly higher than those in the ADSC group(P<0.01).Serum HA and VEGF in ADSC group and ADSC+PRP group were significantly higher than those in control group at 3 d,5 d and 7 d after modeling(P<0.01).Serum HA and VEGF were significantly higher in the ADSC+PRP group than in the ADSC group(P<0.01).Conclusion:Allogeneic PRP combined with ADSC can promote the regeneration and healing of skin wounds in mice.The mechanism may be related to the up-regulation of the expression of vascular endothelial growth factor and HA and the promotion of the formation of blood vessels and granulation tissue.展开更多
miRNAs play important roles in the post-transcriptional regulation of most transcripts and control several biological processes. We have shown that miR-200b regulates VEGF and miR-146a regulates ECM （extracellular ma...miRNAs play important roles in the post-transcriptional regulation of most transcripts and control several biological processes. We have shown that miR-200b regulates VEGF and miR-146a regulates ECM （extracellular matrix） protein, FN （fibronectin） production. We examined the effects of these two miRNAs in wound healing in vitro and in animals with or without diabetes. Microvascular ECs （endothelial cells） with or without miR inhibitor （antagomir） transfection were tested for VEGF and FN production. Scratch assays and angiogenesis assays were performed. Wounds in the back of mice with or without streptozotocin-induced diabetes were treated with antagomirs alone or in combinations. The wounds were measured and tissues were examined for mRNAs, proteins and miRNAs and examined histologically. In the ECs, miR-200b or miR-146a inhibitors increased VEGF and FN production, cell migration and tube formation. Wound healing in the animals, along with increased production of specific proteins were accelerated by miR-200b or miR-146a inhibitor treatment and was pronounced when these two were combined In diabetes, although delayed, similar patterns were seen. These results indicated that combination of miR-146a and miR-200b inhibitor treatment is useful in wound healing both in normal and in diabetic conditions, miRNA mediated wound healing may potentially constitute a novel therapeutic approach.展开更多
基金supported by grants from the National Natural Science Foundation of China(32360138,32060212,32301054 and 81760648)Key Program of Yunnan Fundamental Research Project(202301AS070036)+1 种基金Outstanding Youth Program of Yunnan Applied Basic Research Project-Kunming Medical University Union Foundation(202301AY070001-301)Project of Yunnan Applied Basic Research Project-Kunming Medical University Union Foundation(202101AY070001-006 and 202101AY070001-036).
文摘Background:Wound management of diabetic foot ulcers(DFUs)is a complex and challenging task,and existing strategies fail to meet clinical needs.Therefore,it is important to develop novel drug candidates and discover new therapeutic targets.However,reports on peptides as molecular probes for resolving issues related to DFUs remain rare.This study utilized peptide RL-QN15 as an exogenous molecular probe to investigate the underlying mechanism of endogenous non-coding RNA in DFU wound healing.The aim was to generate novel insights for the clinical management of DFUs and identify potential drug targets.Methods:We investigated the wound-healing efficiency of peptide RL-QN15 under diabetic con-ditions using in vitro and in vivo experimental models.RNA sequencing,in vitro transfection,quantitative real-time polymerase chain reaction,western blotting,dual luciferase reporter gene detection,in vitro cell scratches,and cell proliferation and migration assays were performed to explore the potential mechanism underlying the promoting effects of RL-QN15 on DFU repair.Results:Peptide RL-QN15 enhanced the migration and proliferation of human immortalized keratinocytes(HaCaT cells)in a high-glucose environment and accelerated wound healing in a DFU rat model.Based on results from RNA sequencing,we defined a new microRNA(miR-4482-3p)related to the promotion of wound healing.The bioactivity of miR-4482-3p was verified by inhibiting and overexpressing miR-4482-3p.Inhibition of miR-4482-3p enhanced the migration and proliferation ability of HaCaT cells as well as the expression of vascular endothelial growth factor B(VEGFB).RLQN15 also promoted the migration and proliferation ability of HaCaT cells,and VEGFB expression was mediated via inhibition of miR-4482-3p expression by the p38 mitogen-activated protein kinase(p38MAPK)and smad3 signaling pathways.Conclusions:RL-QN15 is an effective molecule for the treatment of DFUs,with the underlying mechanism related to the inhibition of miR-4482-3p expression via the p38MAPK and smad3 signaling pathways,ultimately promoting re-epithelialization,angiogenesis and wound healing.This study provides a theoretical basis for the clinical application of RL-QN15 as a molecular probe in promoting DFU wound healing.
基金supported by the National Natural Science Foundation of China(No.81141077 and No.81271968)
文摘In order to investigate the promoting effect of low-intensity treadmill exercise on rat dorsal wound healing and the mechanism, 20 Sprague-Dawley rats were randomly divided into two groups： exercise group（Ex） and non-exercise group（non-ex）. The rats in Ex group were given treadmill exercise for one month, and those in non-ex group raised on the same conditions without treadmill exercise. Both groups received dorsal wound operation with free access to food and water. By two-week continuous observation and recording of the wound area, the healing rate was analyzed. The blood sample was collected at day 14 post-operation via cardiac puncture for determination of the number of endothelial progenitor cells（EPCs） by flow cytometry, and the concentrations of relevant cytokines such as basic fibroblast growth factor（b FGF）, endothelial nitric oxide synthase（e NOS） and vascular endothelial growth factor（VEGF） were measured by ELISA. The skin tissue around the wound was dissected to observe the vascular density under the microscope after HE staining, to detect the m RNA level of VEGFR2 and angiopoietin-1（Ang-1） receptor using RT-q PCR, and protein expression of α-smooth muscle actin（αSMA） and type collagenⅢ（ColⅢ） using Western blotting. It was found that the wound area in Ex group was smaller at the same time point than in non-ex group. The number of circulating EPCs was greater and the concentrations of vasoactive factors such as VEGF, e NOS and b FGF were higher in Ex group than in non-ex group. HE staining displayed a higher vessel density in Ex group than in non-ex group. Moreover, the m RNA expression of VEGFR2 and Ang-1 detected in the wound tissue in Ex group was higher than in non-ex group. Meanwhile, the protein expression of αSMA and Col Ⅲwas more abundant in Ex group than in non-ex group. Conclusively, the above results demonstrate Ex rats had a higher wound healing rate, suggesting low-intensity treadmill exercise accelerates wound healing. The present work may provide some hint for future study of treating refractory wound.
基金National Natural Science Foundation of China.Project No:81301568.
文摘Objective:Elet-rich plasma(PRP)combined with adipose-derived mesenchymal stem cells(ADSC)on skin wound healing and serum hyaluronic acid(HA)and vascular endothelial growth factor(VEGF)levels in mice.Methods:Forty-five mice were randomly divided into three groups(n=15),which were control,ADSC,and ADSC+PRP.The wound injury model was established in all three groups.The control group was injected subcutaneously with normal saline,the ADSC group was injected subcutaneously with allogeneic PRP,and the ADSC+PRP was injected subcutaneously with ADSC and PRP.Wound healing was observed at 3 d,5 d,and 7 d after modeling,and histomorphological features were observed by HE staining.CollagenⅠand CollagenⅢlevels were detected using Western blot.The levels of serum HA and VEGF in each group were detected and compared.Results:The wound healing rate of ADSC group and ADSC+PRP group was significantly higher than that of control group at 3 d,5 d and 7 d after modeling(P<0.01).The wound healing rate of the ADSC+PRP group was significantly higher than that of the ADSC group(P<0.01).The inflammatory response of the control group and the ADSC group was significant at 3 and 5 d after modeling,and there was a small amount of granulation tissue in the ADSC group.The ADSC+PRP group had a milder inflammatory response and more granulation tissue;After 7 d,the ADSC+PRP group had more neovascularization and higher re-epithelialization rate.The control group and the ADSC group had poorer epithelialization and fewer new blood vessels.The expression levels of CollagenⅠand CollagenⅢin ADSC group and ADSC+PRP group were significantly higher than those in control group at 3 d,5 d and 7 d after modeling(P<0.01).CollagenⅠand CollagenⅢin the ADSC+PRP group were significantly higher than those in the ADSC group(P<0.01).Serum HA and VEGF in ADSC group and ADSC+PRP group were significantly higher than those in control group at 3 d,5 d and 7 d after modeling(P<0.01).Serum HA and VEGF were significantly higher in the ADSC+PRP group than in the ADSC group(P<0.01).Conclusion:Allogeneic PRP combined with ADSC can promote the regeneration and healing of skin wounds in mice.The mechanism may be related to the up-regulation of the expression of vascular endothelial growth factor and HA and the promotion of the formation of blood vessels and granulation tissue.
文摘miRNAs play important roles in the post-transcriptional regulation of most transcripts and control several biological processes. We have shown that miR-200b regulates VEGF and miR-146a regulates ECM （extracellular matrix） protein, FN （fibronectin） production. We examined the effects of these two miRNAs in wound healing in vitro and in animals with or without diabetes. Microvascular ECs （endothelial cells） with or without miR inhibitor （antagomir） transfection were tested for VEGF and FN production. Scratch assays and angiogenesis assays were performed. Wounds in the back of mice with or without streptozotocin-induced diabetes were treated with antagomirs alone or in combinations. The wounds were measured and tissues were examined for mRNAs, proteins and miRNAs and examined histologically. In the ECs, miR-200b or miR-146a inhibitors increased VEGF and FN production, cell migration and tube formation. Wound healing in the animals, along with increased production of specific proteins were accelerated by miR-200b or miR-146a inhibitor treatment and was pronounced when these two were combined In diabetes, although delayed, similar patterns were seen. These results indicated that combination of miR-146a and miR-200b inhibitor treatment is useful in wound healing both in normal and in diabetic conditions, miRNA mediated wound healing may potentially constitute a novel therapeutic approach.
