简介：无

简介：AbstractPurpose:The proposed pathological mechanism for scar formation is controversial, and increased attention has been paid to the fatty acids (FAs) in the formation of pathological scars. Notably, FAs are known to be important in inflammation and mechanotransduction, which is closely related to scar formation. Therefore, it is necessary to clarify the roles of FA in scar formation.Methods:Hypertrophic scar and keloid formed for more than a year and without other treatment, as well as normal skin samples were obtained from patients who underwent plastic surgery. Finally, keloids (n = 10), hypertrophic scars (n = 10), and normal skin samples (n = 10) were collected under informed consent. Primary dermal fibroblasts were isolated and cultured. The amount and variety of FAs were detected by lipid chromatography-mass spectrometry. Immunohistochemistry, real-time PCR, and western blotting were used to verify the expression of sterol regulatory element-binding protein-1 (SREBP1) and fatty acid synthase (FASN) in the samples and their fibroblasts. Student's t-test, ANOVA, and orthogonal partial least square discriminant analysis were performed for statistical analysis (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).Results:Compared with full-thickness normal skin, there were 27 differential FAs in keloids and 15 differential FAs in hypertrophic scars (*p < 0.05 and variable influence on projection >1.0). The expression of SREBP1 and FASN was lower in pathological scars both at mRNA and protein levels (all*p < 0.05). However, the mRNA levels of SREBP1 (***p = 0.0002) and FASN (***p = 0.0021) in keloid-derived fibroblasts were higher than that in normal skin fibroblasts (NFBs), while the expression in hypertrophic scar-derived fibroblasts was lower than that in NFBs (both *p < 0.05). Whereas there was no significant difference in FASN protein expression between keloid-derived fibroblasts and NFBs (p > 0.05).Conclusion:FAs involved in pathological scars are abnormally changed in scar formation. Thus, fatty acid-derived inflammation and de novo synthesis pathway of FA may play a key role in the formation of pathological scars.

简介：AbstractObjectives:Mechanistic target of rapamycin (mTOR) activation has been identified in keloid. This study aimed to identify the role of mTOR-dependent autophagy activity in keloid.Methods:We detected the expression of specific proteins representing mTOR activity and baseline autophagy levels in keloid tissues (KTs) and primary keloid fibroblasts (KFs) using immunohistochemical staining and western blotting. Simultaneously, the formation of acid vesicles was assessed by acridine orange staining in KFs. To investigate whether mTOR-dependent pathway mediated the regulation of autophagy machinery in keloid, we first validated whether mTOR inhibitors, rapamycin (100 nmol/L) and KU-0063794 (5 μmol/L), could inhibit mTOR activity in KFs by western blotting. Then we explored the reverse effects on autophagy activity induced by mTOR inhibitors in the presence of lysosomal protease inhibitors by western blotting.Results:It demonstrated elevated expression of mTOR, S6, and their activated forms in KTs, and an elevated expression of p-S6 Ser235/236 in KFs, suggesting mTOR was activated in keloid. Less LC3 and Beclin1 were expressed in the cytoplasm of KFs, whereas Ubiquitin was abundantly expressed in KTs compared with extra-lesional tissues. In addition, at the cellular level, an impeded conversion of LC3-I to LC3-II was shown in KFs and the formation of acid vesicles were also decreased in KFs compared with normal fibroblasts (NFs), indicating that autophagy activity is defective in keloid. mTOR inhibitors, Rapamycin (E-64d + pepstatin vs. rapamycin + E-64d + pepstatin: [0.88 ± 0.35] vs. [1.56 ± 0.46], F= 5.56, P= 0.049) and KU-0063794 (E-64d + pepstatin vs. KU-0063794 + E-64d + pepstatin: [0.92 ± 0.22] vs. [1.51 ± 0.25], F= 25.88, P= 0.011) can reverse the inhibition effect on autophagy of KFs while inhibiting mTOR activity.Conclusion:Autophagy machinery is inhibited in keloid which is regulated by mTOR-dependent pathway.

简介：无

简介：Theresearchgroupwasinterestedinthetherapeuticpotentialoflimbalfibroblasts,whicharepre-sentinthecorneallimbus.Recently,inmuchstemcellresearch,researchersandadvocatesarestress-ingtheimportanceofutilizingcellsfromthenichewhichtheywilltarget.Thismeans,thatadiposetissueandbonemarrowderivedstemcells,arelikelynotappropriateandevendangeroustoapplyu-biquitouslyastherapiestootherpartsofthebodyincertaincases.Severalresearchgroupsandclini-caltrialshaveshownthistobethecase,includingcasesofblindness,braininflammation,anddeathasadverseevents.

简介：AnargonatmosphericpressureplasmajetwasemployedtotreatL929murinefibroblastsculturedinvitro.Experimentalresultsshowedthat,comparedwiththecontrolcells,thetreatmentoffibroblastswith15sofplasmaledtoasignificantincreaseofcellviabilityandcollagensynthesis,whilethetreatmentof25splasmaresultedinaremarkabledecrease.Explorationofrelatedmechanismssuggestedthatcoldplasmacouldup-regulateCyclinD1geneexpressionanddown-regulatep27geneexpressionatalowdose,whileitcoulddown-regulateCyclinD1expressionandup-regulatep27expressionatahigherdose,thusalteringthecellcycleprogression,andthenaffectingcellviabilityandcollagensynthesisoffibroblasts.

简介：

简介：AIMTo在真菌的部件zymosan在.METHODSHCFs是的有教养的人的角膜的成纤维细胞(HCF)导致的proinflammatorycytokine和chemokine表示上调查triptolide的效果在zymosan或triptolide的缺席或存在有教养。interleukin(IL)的版本-6,IL-8，和进文化上层清液的单核白血球chemoattractantprotein-1(MCP-1)与连接酶的immunosorbent试金被测量。细胞的许多为这些蛋白质的mRNAs被反向的抄写和即时聚合酶链反应分析决定。激活mitogen的蛋白质kinases(MAPK)和内长的原子factor-κ的phosphorylation；B(NF-κ；B)禁止者Iκ；B-α；被immunoblot分析检验。版本喂奶从HCF的脱氢酶(LDH)活动被测量，比色的assay.RESULTSTriptolide禁止了从在一个集中依赖者和时间依赖者举止的HCF的IL-6，IL-8，和MCP-1的导致zymosan的版本。它也在这些房间禁止了IL-6，IL-8，和MCP-1mRNA丰富的导致zymosan的起来规定。而且，triptolide稀释了MAPK的导致zymosan的phosphorylation细胞外的调整信号的kinase(英皇家空军之阶级最低之兵)，c6月NH2-terminalkinase(JNK)，和象Iκ的phosphorylation和降级一样的p38；B-α；。Triptolide没多半展出cytotoxicity因为HCFs.CONCLUSIONTriptolide由暴露于zymosan的HCF禁止了proinflammatorycytokine和chemokine生产，随这个行动被MAPK和NF-κ的抑制调停；B发信号小径。这混合物可能因此被期望限制煽动性的房间的渗入进与真菌的感染联系的角膜。

简介：AbstractBackground:Fibrosis in the peripheral airways contributes to airflow limitation in patients with chronic obstructive pulmonary disease (COPD). However, the key proteins involved in its development are still poorly understood. Thus, we aimed to identify the differentially expressed proteins (DEPs) between smoker patients with and without COPD and elucidate the molecular mechanisms involved by investigating the effects of the identified biomarker candidate on lung fibroblasts.Methods:The potential DEPs were identified by isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic analysis. The messenger RNA and protein levels of clusterin (CLU) in COPD patients and 12% cigarette smoke extract (CSE)-treated human bronchial epithelial cells were determined at the indicated time points. Furthermore, an in vitro COPD model was established via the administration of 8% CSE to normal human lung fibroblasts (NHLFs) at indicated time points. The effects of CSE treatment and CLU silencing on proliferation and activation of lung fibroblasts were analyzed.Results:A total of 144 DEPs were identified between COPD patients and normal smokers. The iTRAQ-based proteomics and bioinformatics analyses identified CLU as a serum biomarker candidate. We also discovered that CLU levels were significantly increased (P < 0.0001) in Global Initiative for Obstructive Lung Disease II, III, and IV patients and correlated (P < 0.0001) with forced expiratory volume in 1 s (R=-0.7705), residual volume (RV) (R = 0.6281), RV/total lung capacity (R = 0.5454), and computerized tomography emphysema (R = 0.7878). Similarly, CLU levels were significantly increased in CSE-treated cells at indicated time points (P < 0.0001). The CSE treatment significantly inhibited the proliferation, promoted the inflammatory response, differentiation of NHLFs, and collagen matrix deposition, and induced the apoptosis of NHLFs; however, these effects were partially reversed by CLU silencing.Conclusion:Our findings suggest that CLU may play significant roles during airway fibrosis in COPD by regulating lung fibroblast activation.

简介：Aim:ToobservetheapoptoticchangesfollowingexposuretoEMPandtoexplorethepossibleinjurymechanisminNIH-3T3fibroblasts.Methods:FollowingNIH-3T3cellswereexposedtoEMP,theproliferationandviabilityofNIH-3T3fibroblastswereestimatedbyhemacytometerandMTTMeasurements.Apoptoticratewasmeasuredbyflowcytometer.TheimnmohistochemicalSPmethodwasusedtodeterminebcl-2,p53.ThepositivelystainedcellswereanalyzedwithCMIAS-Ⅱimageanalysissystematamagnification400.AlldatawereanalyzedbySPSS8.0software.Results:TheproliferationandviabilityofNIH-3T3cellsweremarkedlyinhibitedandsignificantapoptosiswasinducedfollowingexposuretoEMP.EMPcouldincreasethenumberofnon-adherentcells,thehighestratio(33.9%)ofnon-adherentcellsalsooccurredat6h.TheAs70valueofMTTdecreasedimmediatelyat1h,6hfollowingthecellswereexposedascomparedwiththecontrol(P＜0.05).Thehighestratioofapoptosiswas15.07%at6h,thendecreasedgradually.Down-regulationofbcl-2andup-regulationofp53wereinducedbyEMP(P＜0.05).Conclusions:EMPcouldpromoteapoptosisofNIH-3T3fibroblasts.EMPcouldalsodown-regulatebcl-2levelandup-regulatep53levelinNIH-3T3fibroblasts.Bcl-2andp53genemaytakepartintheprocessofapoptosis.

简介：AIMTo与人的角膜的上皮的房间和成纤维细胞调查角膜的前面的薄片状的重建的可行性，并且在vitro.METHODSThe支架的一个非细胞组成的猪的角膜矩阵(APCM)从与0.5%钠dodecyl硫酸盐(SDS)被对待的新鲜猪的角膜被准备答案和角膜的房间的完全的移动被hematoxylin曙红证实(他)染色并且4′；，染色的6-diamidino-2-phenylindole(DAPI)。人的角膜的成纤维细胞和上皮的房间与沥滤是有教养的从APCM提取的液体，然后房间proliferative能力被MTT评估试金。构造人的角膜的前面的薄片状的代替，角膜的成纤维细胞被注入APCM并且为3d有教养，由culturing列在后面在基质构造的角膜的上皮的房间为另一10d出现。角膜的代替被分析由他染色，并且immunofluorescencestaining.RESULTSHistological检查显示在APCM由没有房间他染色，并且染色的DAPI没检测任何剩余DNA。从APCM的沥滤的液体几乎没在人的角膜的成纤维细胞和上皮的房间的增长能力上有小影响。在10d，一连续盖住APCM的表面的人的角膜的上皮的房间的3～5层被观察，并且注射角膜的成纤维细胞在支架以内散布了。构造的显型类似于正常人的角膜，与cytokeratin的高表示12在上皮的房间层和在stroma.CONCLUSIONCorneal的vimentin的高表示，前面的薄片状的代替能被与一个非细胞组成的猪的角膜矩阵栽培人的角膜的上皮的房间和成纤维细胞在vitro重建。这在vivo为进一步的移植打了基础。

简介：Objective:ToinvestigatetheeffectofsubstanceP(SP)ongeneexpressionoftransforminggrowthfactorβ-1(TGFβ-1),transforminggrowthfactorreceptor-1(TGFR-1)andtransforminggrowthfactorreceptor-2(TGFR-2)infibroblastsculturedinvitrofromrat'sgranulationtissues.Methods:Thefibroblastsfromthegranulationtissuesintheskeletalmuscleofrat'shindlimbsinjuredbyformaldehydewereculturedinvitro.Whendifferentconcentrations(10-9-10-5mol/L)ofSPwereaddedintotheculturemedium,thechangesofgeneexpressionofTGFβ-1,TGFR-1andTGFR-2intheculturedfibroblastswereobservedwithreversetranscriptionpolymerasechainreactionatdifferentintervals(0,3,6,12and24hoursafterincubation).Results:ThegeneexpressionofTGFβ-1,TGFR-1andTGFR-2inthefibroblastsculturedfromrat'sgranulationtissueswasup-regulatedbySP.ThepeaklevelofthemRNAexpressionwasfoundat10-8mol/LSPandtheup-regulationeffectwasnotfoundat10-5mol/Land10-6mol/L.ThepeaklevelsofgeneexpressionofTGFβ-1,TGFR-1andTGFR-2inthefibroblaststreatedwithSPwereachievedat6and12hours,respectively.Conclusions:SPhasup-regulationeffectonthegeneexpressionofTGFβ-1,TGFR-1andTGFR-2infibroblastsfromrat'sgranulationtissuesinvitro,andtheeffectisrelatedtodifferentstimulatingconcentrationsofSP.Itmaybeconcernedwithproliferationanddifferentiationoffibroblastsandformationofscartissuesduringwoundhealing.

简介：现在的学习的目的是系统地在成纤维细胞和造骨细胞粘附和增长上评估聚合物crystallinity的角色用一系列poly(caprolactone-co-glycolide)(打印机控制语言/针网阵列)聚合物。自从他们反映高度水晶、非结晶的材料，打印机控制语言/针网阵列聚合物被选择。打印机控制语言/针网阵列聚合材料被塑造进薄电影的压缩制作。从到在25:75，35:65和45:55的比率的打印机控制语言/针网阵列的中间的copolymeric作文的打印机控制语言或针网阵列，五篇作文被学习。当共聚物是非结晶的时，纯打印机控制语言和针网阵列代表了水晶的材料。聚合物/共聚物用DSC被描绘为nanotopography估计crystallinity，为hydrophobicity的接触角度测量，和AFM。而他们在crystallinity显著地不同，打印机控制语言/针网阵列电影表明了类似的hydrophobicity和nanotopography。对的房间粘附和打印机控制语言/针网阵列电影和增长研究上的增长用造骨细胞和NIH-3T3成纤维细胞被执行。高度水晶、僵硬的打印机控制语言和针网阵列表面是，这被观察显著地在支持成纤维细胞生长更有效，而非结晶/灵活的打印机控制语言/针网阵列35:65是显著地在支持造骨细胞的生长更有效。这研究证明当化学作文，hydrophobicity和打印机控制语言/针网阵列聚合物的表面粗糙被保持经常时，打印机控制语言/针网阵列的crystallinity和刚硬在决定房间回答起了主要作用。

简介：AbstractBackground:There is growing evidence that 5-fluorouracil (5-FU) combined with therapeutic trauma can effectively induce skin repigmentation in vitiligo patients who are unresponsive to conventional treatments. Previous studies have mainly focused on identifying the antimitotic activity of 5-FU for the treatment of skin cancer, but few studies have investigated its extra-genotoxic actions favoring melanocyte recruitment.Methods:We utilized the full thickness excisional skin wound model in Dct-LacZ transgenic mice to dynamically assess the migration of melanocytes in the margins of wounds treated with or without 5-FU. The in-situ expression of CXCL12 was examined in the wound beds using immunofluorescence staining. Quantitative real-time polymerase chain reaction and Western blotting analyses were performed to detect the expression levels of CXCL12 mRNA and protein in primary mouse dermal fibroblasts treated with or without 5-FU. Transwell assays and fluorescein isothiocyanate (FITC)-phalloidin staining were used to observe cell migration and filamentous actin (F-actin) changes of melan-a murine melanocytes.Results:Whole mount and cryosection X-gal staining showed that the cell numbers of LacZ-positive melanocytes were much higher in the margins of dorsal and tail skin wounds treated with 5-FU compared with the controls. Meanwhile, CXCL12 immunostaining was significantly increased in the dermal compartment of wounds treated with 5-FU (control vs. 5-FU, 22.47 ± 8.85 vs. 44.69 ± 5.97, P < 0.05). Moreover, 5-FU significantly upregulated the expression levels of CXCL12 mRNA (control vs. 5-FU, 1.00 ± 0.08 vs. 1.54 ± 0.06, P < 0.05) and protein (control vs. 5-FU, 1.00 ± 0.06 vs. 2.93 ± 0.10, P < 0.05) in cultured fibroblasts. Inhibition of the CXCL12/CXCR4 axis suppressed melanocyte migration in vitro using a CXCL12 small interfering RNA (siRNA) or a CXCR4 antagonist (AMD3100).Conclusion:5-FU possesses a pro-pigmentary activity through activation of the CXCL12/CXCR4 axis to drive the chemotactic migration of melanocytes.

简介：AbstractBackground:Macrophages are involved in the pathogenesis of idiopathic pulmonary fibrosis, partially by activating lung fibroblasts. However, how macrophages communicate with lung fibroblasts is largely unexplored. Exosomes can mediate intercellular communication, whereas its role in lung fibrogenesis is unclear. Here we aim to investigate whether exosomes can mediate the crosstalk between macrophages and lung fibroblasts and subsequently induce fibrosis.Methods:In vivo, bleomycin (BLM)-induced lung fibrosis model was established and macrophages infiltration was examined. The effects of GW4869, an exosomes inhibitor, on lung fibrosis were assessed. Moreover, macrophage exosomes were injected into mice to observe its pro-fibrotic effects. In vitro, exosomes derived from angiotensin II (Ang II)-stimulated macrophages were collected. Then, lung fibroblasts were treated with the exosomes. Twenty-four hours later, protein levels of α-collagen I, angiotensin II type 1 receptor (AT1R), transforming growth factor-β (TGF-β), and phospho-Smad2/3 (p-Smad2/3) in lung fibroblasts were examined. The Student’s t test or analysis of variance were used for statistical analysis.Results:In vivo, BLM-treated mice showed enhanced infiltration of macrophages, increased fibrotic alterations, and higher levels of Ang II and AT1R. GW4869 attenuated BLM-induced pulmonary fibrosis. Mice with exosomes injection showed fibrotic features with higher levels of Ang II and AT1R, which was reversed by irbesartan. In vitro, we found that macrophages secreted a great number of exosomes. The exosomes were taken by fibroblasts and resulted in higher levels of AT1R (0.22 ± 0.02 vs. 0.07 ± 0.02, t = 8.66, P = 0.001), TGF-β (0.54 ± 0.05 vs. 0.09 ± 0.06, t= 10.00, P < 0.001), p-Smad2/3 (0.58 ± 0.06 vs. 0.07 ± 0.03, t= 12.86, P < 0.001) and α-collagen I (0.27 ± 0.02 vs. 0.16 ± 0.01, t = 7.01, P = 0.002), and increased Ang II secretion (62.27 ± 7.32 vs. 9.56 ± 1.68, t= 12.16, P < 0.001). Interestingly, Ang II increased the number of macrophage exosomes, and the protein levels of Alix (1.45 ± 0.15 vs. 1.00 ± 0.10, t = 4.32, P = 0.012), AT1R (4.05 ± 0.64 vs. 1.00 ± 0.09, t = 8.17, P = 0.001), and glyceraldehyde-3-phosphate dehydrogenase (2.13 ± 0.36 vs. 1.00 ± 0.10, t = 5.28, P = 0.006) were increased in exosomes secreted by the same number of macrophages, indicating a positive loop between Ang II and exosomes production.Conclusions:Exosomes mediate intercellular communication between macrophages and fibroblasts plays an important role in BLM-induced pulmonary fibrosis.

简介：

简介：