Background:Histological and functional recovery after peripheral nerve injury(PNI)is of significant clinical value as delayed surgical repair and longer distances to innervate terminal organs may account for poor outc...Background:Histological and functional recovery after peripheral nerve injury(PNI)is of significant clinical value as delayed surgical repair and longer distances to innervate terminal organs may account for poor outcomes.Low-intensity extracorporeal shock wave therapy(LiESWT)has already been proven to be beneficial for injured tissue recovery on various pathological conditions.The objective of this study was to explore the potential effect and mechanism of LiESWT on PNI recovery.Methods:In this project,we explored LiESWT’s role using an animal model of sciatic nerve injury(SNI).Shockwave was delivered to the region of the SNI site with a special probe at 3 Hz,500 shocks each time,and 3 times a week for 3 weeks.Rat Schwann cells(SCs)and rat perineurial fibroblasts(PNFs)cells,the two main compositional cell types in peripheral nerve tissue,were cultured in vitro,and LiESWT was applied through the cultured dish to the adherent cells.Tissues and cell cultures were harvested at corresponding time points for a reverse transcription-polymerase chain reaction,Western blotting,and immunofluorescence staining.Multiple groups were compared by using one-way analysis of variance followed by the Tukey-Kramer test for post hoc comparisons.Results:LiESWT treatment promoted the functional recovery of lower extremities with SNI.More nerve fibers and myelin sheath were found after LiESWT treatment associated with local upregulation of mechanical sensitive yes-associated protein(YAP)/transcriptional co-activator with a PDZ-binding domain(TAZ)signaling pathway.In vitro results showed that SCs were more sensitive to LiESWT than PNFs.LiESWT promoted SCs activation with more expression of p75(a SCs dedifferentiation marker)and Ki67(a SCs proliferation marker).The SCs activation process was dependent on the intact YAP/TAZ signaling pathway as knockdown of TAZ by TAZ small interfering RNA significantly attenuated this process.Conclusion:The LiESWT mechanical signal perception and YAP/TAZ upregulation in SCs might be one of the underlyi展开更多
This study aimed to compare the effects of bilateral cavernous nerve crushing(BCNC)and bilateral cavernous nerve resection(BCNR)on intracavernous pressure(ICP)and cavernous pathology in rats and to explore the optimal...This study aimed to compare the effects of bilateral cavernous nerve crushing(BCNC)and bilateral cavernous nerve resection(BCNR)on intracavernous pressure(ICP)and cavernous pathology in rats and to explore the optimal treatment time for the BCNC and BCNR models.Seventy-two male rats aged 12 weeks were randomly divided into three equal groups:Sham(both cavernous nerves exposed only),BCNC(BCN crushed for 2 min),and BCNR(5 mm of BCN resected).Erectile function was then measured at 1 week,3 weeks,and 5 weeks after nerve injury,and penile tissues were harvested for histological and molecular analyses by immunohistochemistry,immuno fluoresce nee,Western blot,and cytokine array.We found that erectile function parameters including the maximum,area,and slope of ICP/mean arterial pressure(MAP)significantly decreased after BCNR and BCNC at 1 week and 3 weeks.At 5 weeks,no significant differences were observed in ICP/MAP between the BCNC and Sham groups,whereas the ICP/MAP of the BCNR group remained significantly lower than that of the Sham group.After BCNC and BCNR,the amount of neuronal-nitric oxide synthase-positive fibers,smooth muscle cells,and endothelial cells decreased,whereas the amount of collage n III con tent increased.These pathological cha nges recovered over time,especially in the BCNC group.Our fin dings demonstrate that BCNC leads to acute and reversible erectile dysfunction,thus treatme nt time should be restricted to the first 3 weeks post-BCNC.In contrast,the self-healing ability of the BCNR model is poor,making it more suitable for long-term treatment research.展开更多
BACKGROUND Myocardial ischemia-reperfusion(I/R) is a serious and irreversible injury. Bone marrow-derived mesenchymal stem cells(MSCs) is considered to be a potential therapy for I/R injury due to the paracrine effect...BACKGROUND Myocardial ischemia-reperfusion(I/R) is a serious and irreversible injury. Bone marrow-derived mesenchymal stem cells(MSCs) is considered to be a potential therapy for I/R injury due to the paracrine effects. High-mobility group box1(HMGB1) is a novel mediator in MSC and regulates the response of inflammation injury. Signal Transduction and Transcription Activator 3(STAT3) is a critical transcription factor and important for release of paracrine factors. However, the relationship between HMGB1 and STAT3 in paracrine effect of MSC remains unknown.METHODS In vitro, hypoxia/reoxygenation injury model was established by Anaero Pack System and examined by Annexin V flow cytometry, CCK8 assay and morphology observation. Detection of apoptotic proteins and protein expression of HMGB1and STAT3 by Western blot.RESULTS The conditioned medium of MSCs with or without LPS pretreatment was cocultured with H9C2 cells for 24 h before hypoxia treatment and MSC showed obvious cardiomyocytes protect role, as evidence by decreased apoptosis rate and improved cells viability, and LPS pretreated MSC exhibited better protect role than untreated MSC. However, such effect was abolished in HMGB1 deficiency group, silencing HMGB1 decreased the secretion of vascular endothelial growth factor(VEGF), hepatocyte growth factor(HGF), insulin growth factor(IGF), cell viability, and the expression of STAT3. Furthermore, STAT3 silence attenuated the protective effect of LPS in MSC.CONCLUSIONS These findings suggested that LPS improved MSC-mediated cardiomyocytes protection by HMGB1/STAT3signaling.展开更多
基金the National Natural Science Foundation of China(No.81701434).
文摘Background:Histological and functional recovery after peripheral nerve injury(PNI)is of significant clinical value as delayed surgical repair and longer distances to innervate terminal organs may account for poor outcomes.Low-intensity extracorporeal shock wave therapy(LiESWT)has already been proven to be beneficial for injured tissue recovery on various pathological conditions.The objective of this study was to explore the potential effect and mechanism of LiESWT on PNI recovery.Methods:In this project,we explored LiESWT’s role using an animal model of sciatic nerve injury(SNI).Shockwave was delivered to the region of the SNI site with a special probe at 3 Hz,500 shocks each time,and 3 times a week for 3 weeks.Rat Schwann cells(SCs)and rat perineurial fibroblasts(PNFs)cells,the two main compositional cell types in peripheral nerve tissue,were cultured in vitro,and LiESWT was applied through the cultured dish to the adherent cells.Tissues and cell cultures were harvested at corresponding time points for a reverse transcription-polymerase chain reaction,Western blotting,and immunofluorescence staining.Multiple groups were compared by using one-way analysis of variance followed by the Tukey-Kramer test for post hoc comparisons.Results:LiESWT treatment promoted the functional recovery of lower extremities with SNI.More nerve fibers and myelin sheath were found after LiESWT treatment associated with local upregulation of mechanical sensitive yes-associated protein(YAP)/transcriptional co-activator with a PDZ-binding domain(TAZ)signaling pathway.In vitro results showed that SCs were more sensitive to LiESWT than PNFs.LiESWT promoted SCs activation with more expression of p75(a SCs dedifferentiation marker)and Ki67(a SCs proliferation marker).The SCs activation process was dependent on the intact YAP/TAZ signaling pathway as knockdown of TAZ by TAZ small interfering RNA significantly attenuated this process.Conclusion:The LiESWT mechanical signal perception and YAP/TAZ upregulation in SCs might be one of the underlyi
基金This work was supported by the National Natural Science Foundation of China(No.81671450).
文摘This study aimed to compare the effects of bilateral cavernous nerve crushing(BCNC)and bilateral cavernous nerve resection(BCNR)on intracavernous pressure(ICP)and cavernous pathology in rats and to explore the optimal treatment time for the BCNC and BCNR models.Seventy-two male rats aged 12 weeks were randomly divided into three equal groups:Sham(both cavernous nerves exposed only),BCNC(BCN crushed for 2 min),and BCNR(5 mm of BCN resected).Erectile function was then measured at 1 week,3 weeks,and 5 weeks after nerve injury,and penile tissues were harvested for histological and molecular analyses by immunohistochemistry,immuno fluoresce nee,Western blot,and cytokine array.We found that erectile function parameters including the maximum,area,and slope of ICP/mean arterial pressure(MAP)significantly decreased after BCNR and BCNC at 1 week and 3 weeks.At 5 weeks,no significant differences were observed in ICP/MAP between the BCNC and Sham groups,whereas the ICP/MAP of the BCNR group remained significantly lower than that of the Sham group.After BCNC and BCNR,the amount of neuronal-nitric oxide synthase-positive fibers,smooth muscle cells,and endothelial cells decreased,whereas the amount of collage n III con tent increased.These pathological cha nges recovered over time,especially in the BCNC group.Our fin dings demonstrate that BCNC leads to acute and reversible erectile dysfunction,thus treatme nt time should be restricted to the first 3 weeks post-BCNC.In contrast,the self-healing ability of the BCNR model is poor,making it more suitable for long-term treatment research.
基金supported by grants from the Natural Science Foundation of Liaoning Province(No.2023-MS-268)Educational Committee Foundation of Liaoning Province(LJKZ0868)+1 种基金Funds for young and middle-aged disciplinary backbones of the Second Hospital of Dalian Medical University(No.dy2yxkgg 202002)partially supported by United Fund of the Second Hospital of Dalian Medical University and Dalian Institute of Chemical physics Chinese Academy of Sciences(No.UF-QN-202005).
文摘BACKGROUND Myocardial ischemia-reperfusion(I/R) is a serious and irreversible injury. Bone marrow-derived mesenchymal stem cells(MSCs) is considered to be a potential therapy for I/R injury due to the paracrine effects. High-mobility group box1(HMGB1) is a novel mediator in MSC and regulates the response of inflammation injury. Signal Transduction and Transcription Activator 3(STAT3) is a critical transcription factor and important for release of paracrine factors. However, the relationship between HMGB1 and STAT3 in paracrine effect of MSC remains unknown.METHODS In vitro, hypoxia/reoxygenation injury model was established by Anaero Pack System and examined by Annexin V flow cytometry, CCK8 assay and morphology observation. Detection of apoptotic proteins and protein expression of HMGB1and STAT3 by Western blot.RESULTS The conditioned medium of MSCs with or without LPS pretreatment was cocultured with H9C2 cells for 24 h before hypoxia treatment and MSC showed obvious cardiomyocytes protect role, as evidence by decreased apoptosis rate and improved cells viability, and LPS pretreated MSC exhibited better protect role than untreated MSC. However, such effect was abolished in HMGB1 deficiency group, silencing HMGB1 decreased the secretion of vascular endothelial growth factor(VEGF), hepatocyte growth factor(HGF), insulin growth factor(IGF), cell viability, and the expression of STAT3. Furthermore, STAT3 silence attenuated the protective effect of LPS in MSC.CONCLUSIONS These findings suggested that LPS improved MSC-mediated cardiomyocytes protection by HMGB1/STAT3signaling.
