Hedgehog信号通路与骨质疏松症

Hedgehog信号通路是一条保守而重要的信号通路,涉及到多种细胞的增殖和分化活动。近年研究发现,Hedgehog信号通路可以通过上调Runx2和Osx等主要转录因子的表达促进间充质干细胞（mesenchymalstemcells,MSCs）向成骨细胞分化,并且抑制MSCs向脂肪细胞分化。Hedgehog信号通路还可以通过调节细胞周期蛋白促进成骨细胞增殖。本文综述总结了Hedgehog信号通路调节成骨细胞增殖分化的作用机制,认为Hedgehog信号通路通过促进成骨细胞增殖分化参与调节骨代谢,为骨质疏松症的治疗提供一种新思路。     

白藜芦醇通过抑制Egr1的表达抑制增生性瘢痕成纤维细胞的生长和迁移

目的 探讨白藜芦醇是否通过调节早期生长反应1(early growth response 1,Egr1)蛋白的表达发挥抑制增生性瘢痕成纤维细胞生长和迁移的作用。方法 选取自2016年1月至2019年1月于北部战区总医院烧伤整形科就诊的12例增生性瘢痕患者作为研究对象,分离培养增生性瘢痕成纤维细胞。将细胞分成4组,分别使用0、0.1、0.5和1.0 g/L的白藜芦醇作用于增生性瘢痕成纤维细胞。处理1～4 d后通过四甲基噻唑蓝（methylthiazolyldiphenyl-tetrazolium bromidesigmam, MTT）实验检测细胞增殖能力。不同因素处理细胞24 h后,流式细胞仪检测细胞周期、细胞凋亡水平,transwell实验检测细胞迁移能力。蛋白免疫印迹实验检测Egr1、Cyclin D1、p21和MMP9的表达水平。结果 细胞增殖实验检测结果显示,白藜芦醇具有显著抑制增生性瘢痕成纤维细胞的增殖能力（P<0.05）。白藜芦醇作用于增生性瘢痕成纤维细胞24 h后,能够显著上调G0-G1期细胞的百分比（P<0.05）,下调S期细胞的百分比（P<0.05）,...     

Hedgehog信号通路在肾间质纤维化中作用的研究进展

肾间质纤维化是慢性肾脏病进展的共同通路。Hedgehog信号通路的激活在其中的作用和在胚胎肾脏发育中的作用似乎一样重要。近年研究发现,异常活化的Hedgehog信号通路与肾间质纤维化发生、发展有着密切联系,降低Hedgehog信号的活化可能是治疗肾间质纤维化的新策略,有望降低慢性肾脏疾病的进展。本文综述Hedgehog信号通路的组成成份和调节,并着重讨论其在肾脏纤维化中的作用。     

结缔组织生长因子影响体外培养瘢痕成纤维细胞ERK／MAPK信号通路活化及增殖的研究

目的探讨结缔组织生长因子（connective tissue growth factor,CTGF）诱导增生性瘢痕（hypertrophic scar,HS）成纤维细胞增殖的信号转导通路。方法采用。H-胸腺嘧啶核苷（^3H—TdR）掺入法观察不同浓度CTGF促HS成纤维细胞增殖的效应,以Western印迹法检测CTGF刺激HS成纤维细胞0、5、10、15、30、60min后,磷酸化及非磷酸化细胞外信号调节激酶1／2（extracellular-signal regulated kinase,ERK1／2）的表达,以两者的比值AI衡量信号通路活化程度;应用特异性阻断剂PD98059阻断ERK通路,MTT法检测CTGF诱导细胞增殖的变化。结果CTGF在一定浓度范围内呈浓度依赖性促HS成纤维细胞增殖,ERK1／2在无CTGF刺激时AI为0．0131±0．0036,CTGF刺激HS成纤维细胞5、10、15、30、60min后,AI值分别为0．0221±0．0033,0．1310±0．0361,0．2090±0．0201,0．1710±0．0379,0．0413±0．0036,在15min达高峰;采用PD98059选择性阻断ERK1／2通路后（A=0．420±0．046）与CTGF刺激组比较（A=0．660±0．035）,细胞增殖显著受到抑制（P〈0．01）。结论ERK1／2是CTGF诱导HS成纤维细胞增殖的主要活化的信号通路。     

结缔组织生长因子影响体外培养瘢痕成纤维细胞ERK/MAPK信号通路活化及增殖的研究

Objective To explore the signal mechanism of proliferation stimulating effect of connec-tive tissue growth factor (CTGF) on hypertrophic scar (HS) derived fibroblasts. Methods <'3>H-TdR in-corporation technique was used to determine the proliferative effect of CTGF at different concentration. Western blot was applied to semi-quantitively analyze the expression of phosphorylated and total ERK1/2 protein after 0, 5, 10, 15, 30, and 60 min of CTGF stimulation, and the relative value of which was de-fined as AI to measure the activation of ERK1/2 signal pathway. PD98059 was admitted to specifically block the ERK1/2 pathway, and subsequently cell proliferation stimulated by CTGF was studied by MTT. Results CTGF could stimulate fibroblasts proliferation with a dose-dependant manner, and activa-ted the ERK1/2 signal pathway, and AI built up to 0.209±0.0201, reaching the apex at 15 min after stimulation performed. Inhibition of ERK1/2 activation by PD98059 suppressed CTGF-mediated HS fi-broblasts proliferation significantly, while OD significantly dropped. Conclusion CTGF induces a prolif-erative response in HS fibroblasts, and this action is mainly dependent on the activation of ERK1/2 signal pathway.     

结缔组织生长因子影响体外培养瘢痕成纤维细胞ERK/MAPK信号通路活化及增殖的研究

Objective To explore the signal mechanism of proliferation stimulating effect of connec-tive tissue growth factor (CTGF) on hypertrophic scar (HS) derived fibroblasts. Methods <'3>H-TdR in-corporation technique was used to determine the proliferative effect of CTGF at different concentration. Western blot was applied to semi-quantitively analyze the expression of phosphorylated and total ERK1/2 protein after 0, 5, 10, 15, 30, and 60 min of CTGF stimulation, and the relative value of which was de-fined as AI to measure the activation of ERK1/2 signal pathway. PD98059 was admitted to specifically block the ERK1/2 pathway, and subsequently cell proliferation stimulated by CTGF was studied by MTT. Results CTGF could stimulate fibroblasts proliferation with a dose-dependant manner, and activa-ted the ERK1/2 signal pathway, and AI built up to 0.209±0.0201, reaching the apex at 15 min after stimulation performed. Inhibition of ERK1/2 activation by PD98059 suppressed CTGF-mediated HS fi-broblasts proliferation significantly, while OD significantly dropped. Conclusion CTGF induces a prolif-erative response in HS fibroblasts, and this action is mainly dependent on the activation of ERK1/2 signal pathway.     

结缔组织生长因子影响体外培养瘢痕成纤维细胞ERK/MAPK信号通路活化及增殖的研究

Objective To explore the signal mechanism of proliferation stimulating effect of connec-tive tissue growth factor (CTGF) on hypertrophic scar (HS) derived fibroblasts. Methods <'3>H-TdR in-corporation technique was used to determine the proliferative effect of CTGF at different concentration. Western blot was applied to semi-quantitively analyze the expression of phosphorylated and total ERK1/2 protein after 0, 5, 10, 15, 30, and 60 min of CTGF stimulation, and the relative value of which was de-fined as AI to measure the activation of ERK1/2 signal pathway. PD98059 was admitted to specifically block the ERK1/2 pathway, and subsequently cell proliferation stimulated by CTGF was studied by MTT. Results CTGF could stimulate fibroblasts proliferation with a dose-dependant manner, and activa-ted the ERK1/2 signal pathway, and AI built up to 0.209±0.0201, reaching the apex at 15 min after stimulation performed. Inhibition of ERK1/2 activation by PD98059 suppressed CTGF-mediated HS fi-broblasts proliferation significantly, while OD significantly dropped. Conclusion CTGF induces a prolif-erative response in HS fibroblasts, and this action is mainly dependent on the activation of ERK1/2 signal pathway.     

结缔组织生长因子影响体外培养瘢痕成纤维细胞ERK/MAPK信号通路活化及增殖的研究

Objective To explore the signal mechanism of proliferation stimulating effect of connec-tive tissue growth factor (CTGF) on hypertrophic scar (HS) derived fibroblasts. Methods <'3>H-TdR in-corporation technique was used to determine the proliferative effect of CTGF at different concentration. Western blot was applied to semi-quantitively analyze the expression of phosphorylated and total ERK1/2 protein after 0, 5, 10, 15, 30, and 60 min of CTGF stimulation, and the relative value of which was de-fined as AI to measure the activation of ERK1/2 signal pathway. PD98059 was admitted to specifically block the ERK1/2 pathway, and subsequently cell proliferation stimulated by CTGF was studied by MTT. Results CTGF could stimulate fibroblasts proliferation with a dose-dependant manner, and activa-ted the ERK1/2 signal pathway, and AI built up to 0.209±0.0201, reaching the apex at 15 min after stimulation performed. Inhibition of ERK1/2 activation by PD98059 suppressed CTGF-mediated HS fi-broblasts proliferation significantly, while OD significantly dropped. Conclusion CTGF induces a prolif-erative response in HS fibroblasts, and this action is mainly dependent on the activation of ERK1/2 signal pathway.     

结缔组织生长因子影响体外培养瘢痕成纤维细胞ERK/MAPK信号通路活化及增殖的研究

Objective To explore the signal mechanism of proliferation stimulating effect of connec-tive tissue growth factor (CTGF) on hypertrophic scar (HS) derived fibroblasts. Methods <'3>H-TdR in-corporation technique was used to determine the proliferative effect of CTGF at different concentration. Western blot was applied to semi-quantitively analyze the expression of phosphorylated and total ERK1/2 protein after 0, 5, 10, 15, 30, and 60 min of CTGF stimulation, and the relative value of which was de-fined as AI to measure the activation of ERK1/2 signal pathway. PD98059 was admitted to specifically block the ERK1/2 pathway, and subsequently cell proliferation stimulated by CTGF was studied by MTT. Results CTGF could stimulate fibroblasts proliferation with a dose-dependant manner, and activa-ted the ERK1/2 signal pathway, and AI built up to 0.209±0.0201, reaching the apex at 15 min after stimulation performed. Inhibition of ERK1/2 activation by PD98059 suppressed CTGF-mediated HS fi-broblasts proliferation significantly, while OD significantly dropped. Conclusion CTGF induces a prolif-erative response in HS fibroblasts, and this action is mainly dependent on the activation of ERK1/2 signal pathway.     

结缔组织生长因子影响体外培养瘢痕成纤维细胞ERK/MAPK信号通路活化及增殖的研究

Objective To explore the signal mechanism of proliferation stimulating effect of connec-tive tissue growth factor (CTGF) on hypertrophic scar (HS) derived fibroblasts. Methods <'3>H-TdR in-corporation technique was used to determine the proliferative effect of CTGF at different concentration. Western blot was applied to semi-quantitively analyze the expression of phosphorylated and total ERK1/2 protein after 0, 5, 10, 15, 30, and 60 min of CTGF stimulation, and the relative value of which was de-fined as AI to measure the activation of ERK1/2 signal pathway. PD98059 was admitted to specifically block the ERK1/2 pathway, and subsequently cell proliferation stimulated by CTGF was studied by MTT. Results CTGF could stimulate fibroblasts proliferation with a dose-dependant manner, and activa-ted the ERK1/2 signal pathway, and AI built up to 0.209±0.0201, reaching the apex at 15 min after stimulation performed. Inhibition of ERK1/2 activation by PD98059 suppressed CTGF-mediated HS fi-broblasts proliferation significantly, while OD significantly dropped. Conclusion CTGF induces a prolif-erative response in HS fibroblasts, and this action is mainly dependent on the activation of ERK1/2 signal pathway.     

结缔组织生长因子影响体外培养瘢痕成纤维细胞ERK/MAPK信号通路活化及增殖的研究

Objective To explore the signal mechanism of proliferation stimulating effect of connec-tive tissue growth factor (CTGF) on hypertrophic scar (HS) derived fibroblasts. Methods <'3>H-TdR in-corporation technique was used to determine the proliferative effect of CTGF at different concentration. Western blot was applied to semi-quantitively analyze the expression of phosphorylated and total ERK1/2 protein after 0, 5, 10, 15, 30, and 60 min of CTGF stimulation, and the relative value of which was de-fined as AI to measure the activation of ERK1/2 signal pathway. PD98059 was admitted to specifically block the ERK1/2 pathway, and subsequently cell proliferation stimulated by CTGF was studied by MTT. Results CTGF could stimulate fibroblasts proliferation with a dose-dependant manner, and activa-ted the ERK1/2 signal pathway, and AI built up to 0.209±0.0201, reaching the apex at 15 min after stimulation performed. Inhibition of ERK1/2 activation by PD98059 suppressed CTGF-mediated HS fi-broblasts proliferation significantly, while OD significantly dropped. Conclusion CTGF induces a prolif-erative response in HS fibroblasts, and this action is mainly dependent on the activation of ERK1/2 signal pathway.     

结缔组织生长因子影响体外培养瘢痕成纤维细胞ERK/MAPK信号通路活化及增殖的研究

Objective To explore the signal mechanism of proliferation stimulating effect of connec-tive tissue growth factor (CTGF) on hypertrophic scar (HS) derived fibroblasts. Methods <'3>H-TdR in-corporation technique was used to determine the proliferative effect of CTGF at different concentration. Western blot was applied to semi-quantitively analyze the expression of phosphorylated and total ERK1/2 protein after 0, 5, 10, 15, 30, and 60 min of CTGF stimulation, and the relative value of which was de-fined as AI to measure the activation of ERK1/2 signal pathway. PD98059 was admitted to specifically block the ERK1/2 pathway, and subsequently cell proliferation stimulated by CTGF was studied by MTT. Results CTGF could stimulate fibroblasts proliferation with a dose-dependant manner, and activa-ted the ERK1/2 signal pathway, and AI built up to 0.209±0.0201, reaching the apex at 15 min after stimulation performed. Inhibition of ERK1/2 activation by PD98059 suppressed CTGF-mediated HS fi-broblasts proliferation significantly, while OD significantly dropped. Conclusion CTGF induces a prolif-erative response in HS fibroblasts, and this action is mainly dependent on the activation of ERK1/2 signal pathway.     

结缔组织生长因子影响体外培养瘢痕成纤维细胞ERK/MAPK信号通路活化及增殖的研究

Objective To explore the signal mechanism of proliferation stimulating effect of connec-tive tissue growth factor (CTGF) on hypertrophic scar (HS) derived fibroblasts. Methods <'3>H-TdR in-corporation technique was used to determine the proliferative effect of CTGF at different concentration. Western blot was applied to semi-quantitively analyze the expression of phosphorylated and total ERK1/2 protein after 0, 5, 10, 15, 30, and 60 min of CTGF stimulation, and the relative value of which was de-fined as AI to measure the activation of ERK1/2 signal pathway. PD98059 was admitted to specifically block the ERK1/2 pathway, and subsequently cell proliferation stimulated by CTGF was studied by MTT. Results CTGF could stimulate fibroblasts proliferation with a dose-dependant manner, and activa-ted the ERK1/2 signal pathway, and AI built up to 0.209±0.0201, reaching the apex at 15 min after stimulation performed. Inhibition of ERK1/2 activation by PD98059 suppressed CTGF-mediated HS fi-broblasts proliferation significantly, while OD significantly dropped. Conclusion CTGF induces a prolif-erative response in HS fibroblasts, and this action is mainly dependent on the activation of ERK1/2 signal pathway.     

结缔组织生长因子影响体外培养瘢痕成纤维细胞ERK/MAPK信号通路活化及增殖的研究

Objective To explore the signal mechanism of proliferation stimulating effect of connec-tive tissue growth factor (CTGF) on hypertrophic scar (HS) derived fibroblasts. Methods <'3>H-TdR in-corporation technique was used to determine the proliferative effect of CTGF at different concentration. Western blot was applied to semi-quantitively analyze the expression of phosphorylated and total ERK1/2 protein after 0, 5, 10, 15, 30, and 60 min of CTGF stimulation, and the relative value of which was de-fined as AI to measure the activation of ERK1/2 signal pathway. PD98059 was admitted to specifically block the ERK1/2 pathway, and subsequently cell proliferation stimulated by CTGF was studied by MTT. Results CTGF could stimulate fibroblasts proliferation with a dose-dependant manner, and activa-ted the ERK1/2 signal pathway, and AI built up to 0.209±0.0201, reaching the apex at 15 min after stimulation performed. Inhibition of ERK1/2 activation by PD98059 suppressed CTGF-mediated HS fi-broblasts proliferation significantly, while OD significantly dropped. Conclusion CTGF induces a prolif-erative response in HS fibroblasts, and this action is mainly dependent on the activation of ERK1/2 signal pathway.     

松果菊苷对人增生性瘢痕成纤维细胞TGF-β1/Smads信号通路影响的实验研究

目的 基于TGF-β1/Smads信号通路探讨松果菊苷(echinacoside,ECH)对人增生性瘢痕成纤维细胞(HSFbs)的作用机制.方法 自2019年3月至2020年2月新疆医科大学第一附属医院整形外科体外培养人增生性瘢痕成纤维细胞;通过CCK-8法分析ECH作用24 h和48 h细胞的抑制活性.采用划痕试验检...     

丹参对增生性瘢痕成纤维细胞形态的影响及增殖抑制初探

目的探索丹参治疗增生性瘢痕的机理。方法将培养的增生性瘢痕成纤维细胞加入含丹参的培养液中培养２４ｈ,与未加药者对照比较。结果①含丹参组成纤维细胞由长梭形变为圆形,而培养上清中的乳酸脱氢酶（ＬＤＨ）活性及ＭＴＴ（四甲基偶氮唑）比色法结果,用药组与对照组均无显著差异（Ｐ＞０．０５）;②流式细胞仪检测各时相细胞ＤＮＡ水平：用药组Ｇ０＋Ｇ１期细胞的百分比明显高于对照组（Ｐ＜０．０１）,而Ｇ２＋Ｍ期百分比则显著低于对照组（Ｐ＜０．０１）。结论丹参能改变增生性瘢痕成纤维细胞形态,而不影响其活力并抑制其增殖。     

骨髓间充质干细胞对瘢痕成纤维细胞作用的机制研究

[目的]观察骨髓间充质干细胞(BM-MSCs)对增生性瘢痕成纤维细胞(HSFB)的影响及其机制研究,为增生性瘢痕的治疗提供实验基础。[方法]分离培养人BM-MSCs和HSFB,制备BM-MSCs条件培养液(CM),分别于12、24和48 h收集CM。将不同时间点收集的CM孵育体外培养的HSFB 24 h,并与空白对照组比较,成纤维细胞的增殖情况采用MTT进行检测,胶原及TGF-β/Smad信号通路的相关基因采用RT-PCR进行检测。[结果]在24、48 h收集的BM-MSCs CM,与对照组相比对HSFB的增殖具有明显的抑制作用(P<0.01),并显著降低了Ⅰ型和Ⅲ型胶原的表达;MSCs在转录水平能显著降低TGF-bRI和TGF-bRII的表达,却能增强Smad7基因的表达,然而,对Smad 2、Smad 3、Smad 4的表达没有影响。[结论]MSCs可以通过对HSFB TGF-β/Smad信号通路的抑制作用,进而达到治疗或抑制增生性瘢痕的目的,为以细胞疗法为治疗策略减轻瘢痕的方法提供了新的理论支持。     

氧化苦参碱通过TGF-β-Smad信号通路调控人增生性瘢痕成纤维细胞增殖及功能

目的 研究氧化苦参碱(OM)对人增生性瘢痕(HS)成纤维细胞增殖、α平滑肌肌动蛋白(α-SM-Actin),Ⅰ、Ⅲ型胶原合成的影响,以及对Smad3和Smad7蛋白表达量的影响,探讨OM对HS成纤维细胞抑制作用的可能机制.方法 体外常规培养HS成纤维细胞,CCK-8检测不同浓度OM对细胞增殖的抑制情况;real-time PCR检测α-SM-Actin、Ⅰ型和Ⅲ型胶原表达;Western blotting检测Smad3和Smad7蛋白量的改变.结果 OM干预后成纤维细胞增殖明显受抑制,抑制率呈现OM浓度依赖性.OM浓度为400 mg/L时,抑制率为53.44%;Ⅰ、Ⅲ型胶原及α-SM-Actin的mRNA相对表达量均明显低于无OM干预组细胞;Smad3蛋白表达下降了48.16%,而Smad7蛋白表达量增加了55.24%.结论 OM作用于HS成纤维细胞可产生抑制效应,其部分机制是通过TGF-β-Smad信号通路的负性调节,实现对细胞的胶原合成及收缩功能的抑制.     

成纤维细胞与创面微环境信号交流失调导致病理性瘢痕形成的机制

病理性瘢痕是因伤口愈合过程发生各种信号交流失调而形成的病理性产物,主要包括增生性瘢痕和瘢痕疙瘩,是整形外科的重难点疾病,严重影响患者的生理和心理健康.近年来研究的病理性瘢痕的病因主要包括遗传基因、细胞因子、炎症递质、免疫应答、环境因素等,各种因素均在病理性瘢痕的发生发展中起一定的作用,但细胞作为生物体的基本单位,必然承...     

雷公藤提取物抑制增生性瘢痕成纤维细胞的实验研究

目的　为雷公藤提取物 (LLZ)治疗烧伤后增生性瘢痕提供实验依据。　方法　体外培养增生性瘢痕成纤维细胞 ,培养液中加入不同浓度的LLZ(5× 10 -3 、5× 10 -4、5× 10 -5、5× 10 -6g/L) ,2 4h后观察细胞形态、增殖活性及药物雷公藤提取物的细胞毒性。　结果　不同浓度的LLZ均能改变成纤维细胞形态 ,减少细胞数量 ,同时可明显降低细胞增殖活性。　结论　LLZ对烧伤后增生性瘢痕成纤维细胞形态和增殖均有明显的抑制作用 ,此作用并非毒性所至。