Background:Sweat glands(SGs)and hair follicles(HFs)are two important cutaneous appendages that play crucial roles in homeostatic maintenance and thermoregulation,and their interaction is involved in wound healing.SGs ...Background:Sweat glands(SGs)and hair follicles(HFs)are two important cutaneous appendages that play crucial roles in homeostatic maintenance and thermoregulation,and their interaction is involved in wound healing.SGs can be regenerated from mesenchymal stem cell-laden 3D bioprinted scaffolds,based on our previous studies,whereas regeneration of HFs could not be achieved in the same model.Due to the lack of an in vitro model,the underlying molecular mechanism of the interaction between SGs and HFs in regeneration could not be fully understood.The purpose of the present study was to establish an in vitro model of skin constructs with SGs and HFs and explore the interaction between these two appendages in regeneration.Methods:To investigate the interaction effects between SGs and HFs during their regeneration processes,a combined model was created by seeding HF spheroids on 3D printed SG scaffolds.The interaction between SG scaffolds and HF spheroids was detected using RNA expression and immunofluorescence staining.The effects of microenvironmental cues on SG and HF regeneration were analysed by altering seed cell types and plantar dermis homogenate in the scaffold.Results:According to this model,we overcame the difficulties in simultaneously inducing SG and HF regeneration and explored the interaction effects between SG scaffolds and HF spheroids.Surprisingly,HF spheroids promoted both SG and HF differentiation in SG scaffolds,while SG scaffolds promoted SG differentiation but had little effect on HF potency in HF spheroids.Specifically,microenvironmental factors(plantar dermis homogenate)in SG scaffolds effectively promoted SG and HF genesis in HF spheroids,no matter what the seed cell type in SG scaffolds was,and the promotion effects were persistent.Conclusions:Our approach elucidated a new model for SG and HF formation in vitro and provided an applicable platform to investigate the interaction between SGs and HFs in vitro.This platform might facilitate 3D skin constructs with multiple appendages and unveil the spatiotemporal molecular program of multiple appendage regeneration.展开更多
Objective:To evaluate the mechanisms underlying the protective effect of Chinese herbal medicine Fructus broussonetiae(FB)in both mouse and cell models of Alzheimer’s disease(AD).Methods:APP/PS1 mice treated with FB ...Objective:To evaluate the mechanisms underlying the protective effect of Chinese herbal medicine Fructus broussonetiae(FB)in both mouse and cell models of Alzheimer’s disease(AD).Methods:APP/PS1 mice treated with FB for 2 months and vehicle-treated controls were run through the Morris water maze and object recognition test to evaluate learning and memory capacity.RNA-Seq,Western blotting,and immunofluorescence staining were also conducted to evaluate the effects of FB treatment on various signaling pathways altered in APP/PS1 mice.To further explore the mechanisms underlying FB’s protective effect,PC-12 cells were treated with Aβ25–35 in order to establish an in vitro model of AD.Results:FB-treated mice showed improved learning and memory capacity on both the Morris water maze and object recognition tests.RNA-seq of hippocampal tissue from APP/PS1 mice showed that FB had effects on multiple signaling pathways,specifically decreasing cell apoptotic signaling and increasing AKT and β-catenin signaling.Similarly,FB up-regulated both AKT and β-catenin signaling in PC-12 cells pre-treated with Aβ25–35,in which AKT positively regulated β-catenin signaling.Further study showed that AKT promoted β-catenin signaling via enhancing β-catenin(Ser552)phosphorylation.Moreover,AKT and β-catenin signaling inhibition both resulted in the attenuated survival of FB-treated cells,indicating the AKT/β-catenin signaling is a crucial mediator in FB promoted cell survival.Conclusions:FB exerted neuroprotective effects on hippocampal cells of APP/PS1 mice,as well as improved cell viability in an in vitro model of AD.The protective actions of FB occurred via the upregulation of AKT/β-catenin signaling.展开更多
基金supported partially by the National Nature Science Foundation of China(81830064,81721092,81701906)the National Key Research and Development Plan(2017YFC1103300)+3 种基金Funds of Chinese PLA General Hospital for Military Medical Inno-vation Research Project(CX19026)the CAMS Innovation Fund for Medical Sciences(CIFMS,2019-I2M-5-059)the Military Medical Research and Development Projects(AWS17J005,2019-126)Fostering Funds of Chinese PLA General Hospital for National Distinguished Young Scholar Science Fund(2017-JQPY-002).
文摘Background:Sweat glands(SGs)and hair follicles(HFs)are two important cutaneous appendages that play crucial roles in homeostatic maintenance and thermoregulation,and their interaction is involved in wound healing.SGs can be regenerated from mesenchymal stem cell-laden 3D bioprinted scaffolds,based on our previous studies,whereas regeneration of HFs could not be achieved in the same model.Due to the lack of an in vitro model,the underlying molecular mechanism of the interaction between SGs and HFs in regeneration could not be fully understood.The purpose of the present study was to establish an in vitro model of skin constructs with SGs and HFs and explore the interaction between these two appendages in regeneration.Methods:To investigate the interaction effects between SGs and HFs during their regeneration processes,a combined model was created by seeding HF spheroids on 3D printed SG scaffolds.The interaction between SG scaffolds and HF spheroids was detected using RNA expression and immunofluorescence staining.The effects of microenvironmental cues on SG and HF regeneration were analysed by altering seed cell types and plantar dermis homogenate in the scaffold.Results:According to this model,we overcame the difficulties in simultaneously inducing SG and HF regeneration and explored the interaction effects between SG scaffolds and HF spheroids.Surprisingly,HF spheroids promoted both SG and HF differentiation in SG scaffolds,while SG scaffolds promoted SG differentiation but had little effect on HF potency in HF spheroids.Specifically,microenvironmental factors(plantar dermis homogenate)in SG scaffolds effectively promoted SG and HF genesis in HF spheroids,no matter what the seed cell type in SG scaffolds was,and the promotion effects were persistent.Conclusions:Our approach elucidated a new model for SG and HF formation in vitro and provided an applicable platform to investigate the interaction between SGs and HFs in vitro.This platform might facilitate 3D skin constructs with multiple appendages and unveil the spatiotemporal molecular program of multiple appendage regeneration.
基金Supported by the National Nature Science Foundation of China(No.81473742)Guangdong Science and Technology Foundation(No.2017A050506007)+1 种基金Shenzhen Major Project of Science and Technology Planning(No.JCYJ20170413161352000)Shenzhen"Sanming Project"Foundation(No.SZSM201612049,No.SZSM201610039)。
文摘Objective:To evaluate the mechanisms underlying the protective effect of Chinese herbal medicine Fructus broussonetiae(FB)in both mouse and cell models of Alzheimer’s disease(AD).Methods:APP/PS1 mice treated with FB for 2 months and vehicle-treated controls were run through the Morris water maze and object recognition test to evaluate learning and memory capacity.RNA-Seq,Western blotting,and immunofluorescence staining were also conducted to evaluate the effects of FB treatment on various signaling pathways altered in APP/PS1 mice.To further explore the mechanisms underlying FB’s protective effect,PC-12 cells were treated with Aβ25–35 in order to establish an in vitro model of AD.Results:FB-treated mice showed improved learning and memory capacity on both the Morris water maze and object recognition tests.RNA-seq of hippocampal tissue from APP/PS1 mice showed that FB had effects on multiple signaling pathways,specifically decreasing cell apoptotic signaling and increasing AKT and β-catenin signaling.Similarly,FB up-regulated both AKT and β-catenin signaling in PC-12 cells pre-treated with Aβ25–35,in which AKT positively regulated β-catenin signaling.Further study showed that AKT promoted β-catenin signaling via enhancing β-catenin(Ser552)phosphorylation.Moreover,AKT and β-catenin signaling inhibition both resulted in the attenuated survival of FB-treated cells,indicating the AKT/β-catenin signaling is a crucial mediator in FB promoted cell survival.Conclusions:FB exerted neuroprotective effects on hippocampal cells of APP/PS1 mice,as well as improved cell viability in an in vitro model of AD.The protective actions of FB occurred via the upregulation of AKT/β-catenin signaling.
