The various fibroproliferative disorders affecting humans have in common excess fibroblast activity and persistent overexpression or dysregulated activity of transforming growth factor beta (TGF-β). Cancer has many s...The various fibroproliferative disorders affecting humans have in common excess fibroblast activity and persistent overexpression or dysregulated activity of transforming growth factor beta (TGF-β). Cancer has many similar characteristics. Antineoplastic drugs can downregulate fibroblast activity and cytokine growth factors. This study evaluates the effect of six antineoplastic drugs on keloid and Dupuytren’s disease fibroblasts. Keloid, normal scar, Dupuytren’s affected palmar fascia, and normal palmar fascia fibroblasts were grown and seeded into Fibroblast Populated Collagen Lattices (FPCLs). The FPCLs were treated with one of six antineoplastic drugs or left untreated as controls. At 7 days, supernatants were extracted from all FPCLs and assayed for expression of Transforming Growth Factor beta (TGF)-β<sub>1</sub> and TGF-β<sub>2</sub>. All six antineoplastic drugs significantly inhibited FPCL contraction in both fibroproliferative conditions compared with the untreated controls (p β<sub>1</sub> and TGF-β<sub>2</sub> expression was downregulated in the supernatants of all FPCLs by the drug exposure. Cytotoxicity did not occur in these studies and was not the reason for the results. Although antineoplastic drugs can have significant side effects when given systemically, these results may be minimized when given to small areas involved in fibroproliferative scarring or when given topically or intralesionally. These in vitro results suggest that antineoplastic drugs may have a utility for treating various fibroproliferative disorders and warrant further investigation.展开更多
Objective To study the function of focal adhesion kinase （FAK） in the formation of hypertrophic scar and its interrelationship with integrin α1. Methods Original fibroblasts from human hypertrophic scar and human n...Objective To study the function of focal adhesion kinase （FAK） in the formation of hypertrophic scar and its interrelationship with integrin α1. Methods Original fibroblasts from human hypertrophic scar and human normal dermis were cultured, and immunocytochemistry was applied to detect localization of expres- sion of FAK and integrin α1 in hypertrophic scar and human normal skin fibroblasts. The expression of integrin α1 was detected before and after FAK antibody blocking hypertrophic scar fibroblasts （HSFB） 48 h later. Meanwhile the collagen synthesis was evaluated by [^3 H]-proline incorporation and HSFB cell proliferation was measured by MTT method. Results The expression of FAK and integrin aI of hypertrophic scar fibroblasts was higher than that of the normal skin fibroblasts significantly （ P 〈 0.01 ）. The expression of integrin α1 was reduced after FAK being blocked （ P 〈 0.01 ）. Meanwhile the collagen synthesis of human scar-derived fibroblasts by [^3H] -proline incor- poration was depressed respectively （ P 〈 0. 01 ）. The cell proliferation was inhibited by using 1：100 and 1：200 FAK antibody with MTI＂ method （ P 〈 0. 01 ）. Conclusion FAK is the key point of signal transmission pathway mediated by integrin α1 , which regulates protein synthesis of integrin α1 , it may play an important role in the proliferation and constriction of hypertrophic scar. FAK antibody can inhibit the collagen synthesis and cell proliferation of hypertrophic scar fibroblasts.展开更多
To investigate the molecular mechanism of extracellular matrix overdeposition in hypertrophic scar tissues and to explore MMPs gene therapy for hypertrophic scar. Methods: Hypertrophic scarderived and normal skin-deri...To investigate the molecular mechanism of extracellular matrix overdeposition in hypertrophic scar tissues and to explore MMPs gene therapy for hypertrophic scar. Methods: Hypertrophic scarderived and normal skin-derived fibroblasts were cultured and a recombinant retrovirus vector containing MMP-3 gene was constructed and then transfected into hypertrophic scar fibroblasts. Expressive level of MMP-3 mRNA was detected by dot blotting, and the activity of MMPs was determined by DNP-peptide.Results: Lower expression of MMP-3 mRNA and fewer DNP-peptide hydrolyzed fragments were observed in hypertrophic scar-derived fibroblasts compared with normal skin-derived fibroblasts. Transfection of MMP-3gene into hypertrophic scar-derived fibroblasts could enhance the expression of MMP-3 mRNA (3. 4 fold)and the de novo capacity to hydrolyze DNP-peptide (2. 1 fold). Conclusion: Overdeposition of extracellular matrix in hypertrophic scar tissue was related to low expression of MMP-3 due to its down-degradation of extracellular matrix. MMP-3 gene transfection could be a better way to treat hypertrophic scars by degrading extracellular matrix.展开更多
Objective To explore the inhibitory effects of He-Ne laser repeated irradiation on the collagen synthesis of cultured scar fibroblasts. Method Cultured fibroblasts derived from hypertrophic scars(HS) were irradiated w...Objective To explore the inhibitory effects of He-Ne laser repeated irradiation on the collagen synthesis of cultured scar fibroblasts. Method Cultured fibroblasts derived from hypertrophic scars(HS) were irradiated with He-Ne laser for 30 minutes at various power densities(10,50,100 and 150 mW/cm2),once a day for 3 consecutive days.In 24 hours after repeated irradiation collagen production and type I procollagen mRNA level of fibroblasts were measured with the incorporation of 3H proline and blot hybridization techniques respectively.Results Collagen synthesis and type I procollagen mRNA level remained unchanged when the laser was irradiated at the power density of 10 mW/cm2 or 50 mW/cm2.Compared with control,collagen synthesis and type I procollagne mRNA level were significantly decreases at the power density of 100 mW/cm2 or 150mW/cm2(P< 0.05).Type I procollagen mRNA level at the power density of 150 mW/cm2 was lower than that at the 100 mW/cm2 (P< 0.05).Conclusion Repeated He-Ne laser irradiation at the power density of 100 mW/cm2 or 150 mW/cm2 can suppress collagen synthesis of cultured fibroblasts in HS.The cause of suppression may be associated with down regulation of type I procollagen mRNA expression.展开更多
Objective To explore the relationship between power density of He -Ne laser with the growth of fibroblasts in hypertroph ic scar (HS).Methods The cultured fibroblasts in HS were i rradiated with He -Ne laser(Wavelenth...Objective To explore the relationship between power density of He -Ne laser with the growth of fibroblasts in hypertroph ic scar (HS).Methods The cultured fibroblasts in HS were i rradiated with He -Ne laser(Wavelenth 632.8nm),various power densities such as50mW/cm 2 ,100mW/cm 2 and 150mW/cm 2 were respectively adopted once a day for 10minutes.After 1,3and 5times o f He -Ne laser for dose rate irradiation separatedly ,the cell count and cell circle analysis were counted and examined by trypa n blue staining and flow cytometry .Results The amount of cell after 1times of 50mW /cm 2 laser irradiation was markedly more than teh control(P<0.01).The amount of cells after 5times of 100mW/cm 2 ,3and 5times of 150mW/cm 2 laser irradiation was less than the controls(P <0.05).The result of cell circle analysis was corresponded with that of the cell count.Conclusion Both stimualtion and inhibition of He -Ne laser on the growth of scar fibroblasts can be obtained with vario us power densities.Power density of He -Ne laser associates with the effects on the growth of scar fibroblasts.展开更多
Objective: To gain the knowledge of expression levels of integrins in hypertrophic scar--derived andnormal skin-derived fibroblasts. Methods: Using anti--β1 α1. α2. α3 and a4 integrin McAbs, the expressions ofinte...Objective: To gain the knowledge of expression levels of integrins in hypertrophic scar--derived andnormal skin-derived fibroblasts. Methods: Using anti--β1 α1. α2. α3 and a4 integrin McAbs, the expressions ofintegrins were detected in hypertrophic scar--derived and normal skin-- derived fibroblasts of passage 5 and 15 byenzyme--linked immunosorbent assay (ELISA ) technique. ResultS: The hypertrophic scar-- derived fibroblastspossessed higher expression levels of integrin subunits than normal skin--derived fibroblasts. After the cells werecultured from passage 5 to passage 15, the decrease range of integrin expression in hypertrophic scar-derived was5. 94%~18. 26%, and 26. 19% ~ 46. 84% in normal skin- derived fibroblasts, showing a statistical difference(P<0. 01). Conclusion: Overexpression of integrins in hypertrophic scar fibroblasts may play an important rolein the hypertrophic scar formation and contemporaneous tissue contracture.展开更多
Objective To explore interaction and interrelationship between TGF - β/Smad signal pathway and Wnt/β - catenin signal pathway in pathogenesis of pathological scar. Methods Three cases of keloid ( K group) ,3 of hype...Objective To explore interaction and interrelationship between TGF - β/Smad signal pathway and Wnt/β - catenin signal pathway in pathogenesis of pathological scar. Methods Three cases of keloid ( K group) ,3 of hyperplastic scar ( H group) and 3 of normal skin ( N group) were selected randomly, and then展开更多
Objective To study the relation of the mRNA and protein expression of CyclinA and p21cip1 in different stages hypertrophic scar fibroblast (FB) with its cell cycle,so as to provide theoretical evidence for interventio...Objective To study the relation of the mRNA and protein expression of CyclinA and p21cip1 in different stages hypertrophic scar fibroblast (FB) with its cell cycle,so as to provide theoretical evidence for intervention therapy of展开更多
背景:增生性瘢痕是以成纤维细胞过度增殖、表皮增厚和角质层功能不良为特征的皮肤纤维化疾病,目前其具体发病机制仍不清楚。目的:基于生物信息学筛选增生性瘢痕相关数据集的核心(Hub)基因及重要信号通路,再用细胞实验加以验证,预测对其...背景:增生性瘢痕是以成纤维细胞过度增殖、表皮增厚和角质层功能不良为特征的皮肤纤维化疾病,目前其具体发病机制仍不清楚。目的:基于生物信息学筛选增生性瘢痕相关数据集的核心(Hub)基因及重要信号通路,再用细胞实验加以验证,预测对其可能有治疗作用的小分子药物。方法:从基因表达综合数据库搜索增生性瘢痕相关的数据集,通过R软件筛选差异表达基因,对差异表达基因进行基因本体论和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Gnomes,KEGG)富集分析,使用String在线平台构建差异表达基因的蛋白质相互作用网络,然后分别利用Cytoscape软件中的Cytohubba和MCODE插件筛选出蛋白质相互作用网络中的关键基因和核心模块,进一步将上述关键基因和构成核心模块的基因求交集得到Hub基因,通过荧光定量PCR验证Hub基因mRNA在人增生性瘢痕与正常皮肤表皮干细胞中的表达差异,并利用人类蛋白图谱中组织学数据验证Hub基因编码蛋白在2种组织中表达量和分布的差异,最后用connectivity map数据库预测针对增生性瘢痕的潜在作用药物。结果与结论:①筛选出的差异表达基因中上调基因102个、下调基因702个,基因本体论和KEGG分析结果显示,富集的信号通路及生物学过程主要涉及紧密连接、花生四烯酸代谢、细胞外基质受体交互、表皮发育和角质化等;②取交集得到8个Hub基因与调控胆固醇代谢的甲羟戊酸途径密切相关,分别是HMGCS1、DHCR7、MSMO1、FDPS、MVK、HMGCR、MVD和ACAT2;③荧光定量PCR结果显示,相比正常皮肤组,增生性瘢痕组HMGCS1、DHCR7、MSMO1、FDPS、HMGCR、MVD和ACAT2 mRNA的表达均显著下降(P<0.05),而MVK mRNA的表达无明显变化(P>0.05);④除MVK外,其余Hub基因编码蛋白在正常皮肤组织中表达水平均高于增生性瘢痕组织(P<0.05);⑤评分排列前10的候选药物包括蛋白激酶A抑制剂(H-89)、丝氨酸蛋白酶抑制剂(Dabigatran-Etexilate)、FLT3抑制剂(舒尼替尼)等,其中白藜芦醇和β-谷甾醇均为植物来源;⑥提示与甲羟戊酸代谢途径密切相关的Hub基因可能通过调控脂质代谢影响表皮结构与功能,这可能是增生性瘢痕的重要发病机制之一,此次研究筛选的小分子化合物可作为治疗增生性瘢痕的候选药物。展开更多
目的:探讨circ-0030042与人第10号染色体缺失的磷酸酶(Phosphatase and tensin homolog deleted on chromosome ten,PTEN)的相互作用关系,并分析其在增生性瘢痕(Hypertrophic scar,HS)患者中对成纤维细胞增殖与迁移的影响及作用机制。方...目的:探讨circ-0030042与人第10号染色体缺失的磷酸酶(Phosphatase and tensin homolog deleted on chromosome ten,PTEN)的相互作用关系,并分析其在增生性瘢痕(Hypertrophic scar,HS)患者中对成纤维细胞增殖与迁移的影响及作用机制。方法:通过circRNA序列和定量聚合酶链反应(PCR技术)检测正常皮肤成纤维细胞(NSFBs)和增生性瘢痕患者成纤维细胞(HSFBs)中circ-0030042的表达。用CCK8检测法检测转染48 h后的HSFBs细胞增殖情况。利用stubRFP-sensGFP-LC3基因转染、流式细胞仪及电子显微镜观察circ-0030042对miR-145/PTEN轴调控VEGF水平的表达。利用生物信息学分析、RNA免疫沉淀、免疫荧光检测等方法,揭示circ-0030042介导HS患者成纤维细胞增殖与迁移的作用机制。结果:circ-0030042在增生性瘢痕中显著上调,过表达时作为VEGF海绵抑制miR-145诱导的成纤维细胞,维持体内稳定性。此外,circ-0030042通过海绵化VEGF水平并阻断其miR-145捕获转录因子(FOXO1)mRNA来影响自噬,而circ-0030042诱导FOXO1的抑制被VEGF水平过表达或circ-0030042结合减少所抵消。过表达circ-0030042对成纤维细胞的增殖抑制与VEGF表达的抑制作用被过表达miR-145部分抵消。结论:干扰circ-0030042通过靶向下调miR-145/PTEN轴进而抑制HSFBs细胞的增殖与迁移,进一步诱导恶性细胞凋亡。展开更多
文摘The various fibroproliferative disorders affecting humans have in common excess fibroblast activity and persistent overexpression or dysregulated activity of transforming growth factor beta (TGF-β). Cancer has many similar characteristics. Antineoplastic drugs can downregulate fibroblast activity and cytokine growth factors. This study evaluates the effect of six antineoplastic drugs on keloid and Dupuytren’s disease fibroblasts. Keloid, normal scar, Dupuytren’s affected palmar fascia, and normal palmar fascia fibroblasts were grown and seeded into Fibroblast Populated Collagen Lattices (FPCLs). The FPCLs were treated with one of six antineoplastic drugs or left untreated as controls. At 7 days, supernatants were extracted from all FPCLs and assayed for expression of Transforming Growth Factor beta (TGF)-β<sub>1</sub> and TGF-β<sub>2</sub>. All six antineoplastic drugs significantly inhibited FPCL contraction in both fibroproliferative conditions compared with the untreated controls (p β<sub>1</sub> and TGF-β<sub>2</sub> expression was downregulated in the supernatants of all FPCLs by the drug exposure. Cytotoxicity did not occur in these studies and was not the reason for the results. Although antineoplastic drugs can have significant side effects when given systemically, these results may be minimized when given to small areas involved in fibroproliferative scarring or when given topically or intralesionally. These in vitro results suggest that antineoplastic drugs may have a utility for treating various fibroproliferative disorders and warrant further investigation.
基金Supported by Shanghai Science Committee Project Fund, China (04JC14085)shanghai Health Burean Project Fund, CHina(054042)
文摘Objective To study the function of focal adhesion kinase （FAK） in the formation of hypertrophic scar and its interrelationship with integrin α1. Methods Original fibroblasts from human hypertrophic scar and human normal dermis were cultured, and immunocytochemistry was applied to detect localization of expres- sion of FAK and integrin α1 in hypertrophic scar and human normal skin fibroblasts. The expression of integrin α1 was detected before and after FAK antibody blocking hypertrophic scar fibroblasts （HSFB） 48 h later. Meanwhile the collagen synthesis was evaluated by [^3 H]-proline incorporation and HSFB cell proliferation was measured by MTT method. Results The expression of FAK and integrin aI of hypertrophic scar fibroblasts was higher than that of the normal skin fibroblasts significantly （ P 〈 0.01 ）. The expression of integrin α1 was reduced after FAK being blocked （ P 〈 0.01 ）. Meanwhile the collagen synthesis of human scar-derived fibroblasts by [^3H] -proline incor- poration was depressed respectively （ P 〈 0. 01 ）. The cell proliferation was inhibited by using 1：100 and 1：200 FAK antibody with MTI＂ method （ P 〈 0. 01 ）. Conclusion FAK is the key point of signal transmission pathway mediated by integrin α1 , which regulates protein synthesis of integrin α1 , it may play an important role in the proliferation and constriction of hypertrophic scar. FAK antibody can inhibit the collagen synthesis and cell proliferation of hypertrophic scar fibroblasts.
文摘To investigate the molecular mechanism of extracellular matrix overdeposition in hypertrophic scar tissues and to explore MMPs gene therapy for hypertrophic scar. Methods: Hypertrophic scarderived and normal skin-derived fibroblasts were cultured and a recombinant retrovirus vector containing MMP-3 gene was constructed and then transfected into hypertrophic scar fibroblasts. Expressive level of MMP-3 mRNA was detected by dot blotting, and the activity of MMPs was determined by DNP-peptide.Results: Lower expression of MMP-3 mRNA and fewer DNP-peptide hydrolyzed fragments were observed in hypertrophic scar-derived fibroblasts compared with normal skin-derived fibroblasts. Transfection of MMP-3gene into hypertrophic scar-derived fibroblasts could enhance the expression of MMP-3 mRNA (3. 4 fold)and the de novo capacity to hydrolyze DNP-peptide (2. 1 fold). Conclusion: Overdeposition of extracellular matrix in hypertrophic scar tissue was related to low expression of MMP-3 due to its down-degradation of extracellular matrix. MMP-3 gene transfection could be a better way to treat hypertrophic scars by degrading extracellular matrix.
文摘Objective To explore the inhibitory effects of He-Ne laser repeated irradiation on the collagen synthesis of cultured scar fibroblasts. Method Cultured fibroblasts derived from hypertrophic scars(HS) were irradiated with He-Ne laser for 30 minutes at various power densities(10,50,100 and 150 mW/cm2),once a day for 3 consecutive days.In 24 hours after repeated irradiation collagen production and type I procollagen mRNA level of fibroblasts were measured with the incorporation of 3H proline and blot hybridization techniques respectively.Results Collagen synthesis and type I procollagen mRNA level remained unchanged when the laser was irradiated at the power density of 10 mW/cm2 or 50 mW/cm2.Compared with control,collagen synthesis and type I procollagne mRNA level were significantly decreases at the power density of 100 mW/cm2 or 150mW/cm2(P< 0.05).Type I procollagen mRNA level at the power density of 150 mW/cm2 was lower than that at the 100 mW/cm2 (P< 0.05).Conclusion Repeated He-Ne laser irradiation at the power density of 100 mW/cm2 or 150 mW/cm2 can suppress collagen synthesis of cultured fibroblasts in HS.The cause of suppression may be associated with down regulation of type I procollagen mRNA expression.
文摘Objective To explore the relationship between power density of He -Ne laser with the growth of fibroblasts in hypertroph ic scar (HS).Methods The cultured fibroblasts in HS were i rradiated with He -Ne laser(Wavelenth 632.8nm),various power densities such as50mW/cm 2 ,100mW/cm 2 and 150mW/cm 2 were respectively adopted once a day for 10minutes.After 1,3and 5times o f He -Ne laser for dose rate irradiation separatedly ,the cell count and cell circle analysis were counted and examined by trypa n blue staining and flow cytometry .Results The amount of cell after 1times of 50mW /cm 2 laser irradiation was markedly more than teh control(P<0.01).The amount of cells after 5times of 100mW/cm 2 ,3and 5times of 150mW/cm 2 laser irradiation was less than the controls(P <0.05).The result of cell circle analysis was corresponded with that of the cell count.Conclusion Both stimualtion and inhibition of He -Ne laser on the growth of scar fibroblasts can be obtained with vario us power densities.Power density of He -Ne laser associates with the effects on the growth of scar fibroblasts.
文摘Objective: To gain the knowledge of expression levels of integrins in hypertrophic scar--derived andnormal skin-derived fibroblasts. Methods: Using anti--β1 α1. α2. α3 and a4 integrin McAbs, the expressions ofintegrins were detected in hypertrophic scar--derived and normal skin-- derived fibroblasts of passage 5 and 15 byenzyme--linked immunosorbent assay (ELISA ) technique. ResultS: The hypertrophic scar-- derived fibroblastspossessed higher expression levels of integrin subunits than normal skin--derived fibroblasts. After the cells werecultured from passage 5 to passage 15, the decrease range of integrin expression in hypertrophic scar-derived was5. 94%~18. 26%, and 26. 19% ~ 46. 84% in normal skin- derived fibroblasts, showing a statistical difference(P<0. 01). Conclusion: Overexpression of integrins in hypertrophic scar fibroblasts may play an important rolein the hypertrophic scar formation and contemporaneous tissue contracture.
文摘Objective To explore interaction and interrelationship between TGF - β/Smad signal pathway and Wnt/β - catenin signal pathway in pathogenesis of pathological scar. Methods Three cases of keloid ( K group) ,3 of hyperplastic scar ( H group) and 3 of normal skin ( N group) were selected randomly, and then
文摘Objective To study the relation of the mRNA and protein expression of CyclinA and p21cip1 in different stages hypertrophic scar fibroblast (FB) with its cell cycle,so as to provide theoretical evidence for intervention therapy of
文摘背景:增生性瘢痕是以成纤维细胞过度增殖、表皮增厚和角质层功能不良为特征的皮肤纤维化疾病,目前其具体发病机制仍不清楚。目的:基于生物信息学筛选增生性瘢痕相关数据集的核心(Hub)基因及重要信号通路,再用细胞实验加以验证,预测对其可能有治疗作用的小分子药物。方法:从基因表达综合数据库搜索增生性瘢痕相关的数据集,通过R软件筛选差异表达基因,对差异表达基因进行基因本体论和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Gnomes,KEGG)富集分析,使用String在线平台构建差异表达基因的蛋白质相互作用网络,然后分别利用Cytoscape软件中的Cytohubba和MCODE插件筛选出蛋白质相互作用网络中的关键基因和核心模块,进一步将上述关键基因和构成核心模块的基因求交集得到Hub基因,通过荧光定量PCR验证Hub基因mRNA在人增生性瘢痕与正常皮肤表皮干细胞中的表达差异,并利用人类蛋白图谱中组织学数据验证Hub基因编码蛋白在2种组织中表达量和分布的差异,最后用connectivity map数据库预测针对增生性瘢痕的潜在作用药物。结果与结论:①筛选出的差异表达基因中上调基因102个、下调基因702个,基因本体论和KEGG分析结果显示,富集的信号通路及生物学过程主要涉及紧密连接、花生四烯酸代谢、细胞外基质受体交互、表皮发育和角质化等;②取交集得到8个Hub基因与调控胆固醇代谢的甲羟戊酸途径密切相关,分别是HMGCS1、DHCR7、MSMO1、FDPS、MVK、HMGCR、MVD和ACAT2;③荧光定量PCR结果显示,相比正常皮肤组,增生性瘢痕组HMGCS1、DHCR7、MSMO1、FDPS、HMGCR、MVD和ACAT2 mRNA的表达均显著下降(P<0.05),而MVK mRNA的表达无明显变化(P>0.05);④除MVK外,其余Hub基因编码蛋白在正常皮肤组织中表达水平均高于增生性瘢痕组织(P<0.05);⑤评分排列前10的候选药物包括蛋白激酶A抑制剂(H-89)、丝氨酸蛋白酶抑制剂(Dabigatran-Etexilate)、FLT3抑制剂(舒尼替尼)等,其中白藜芦醇和β-谷甾醇均为植物来源;⑥提示与甲羟戊酸代谢途径密切相关的Hub基因可能通过调控脂质代谢影响表皮结构与功能,这可能是增生性瘢痕的重要发病机制之一,此次研究筛选的小分子化合物可作为治疗增生性瘢痕的候选药物。
文摘目的:探讨circ-0030042与人第10号染色体缺失的磷酸酶(Phosphatase and tensin homolog deleted on chromosome ten,PTEN)的相互作用关系,并分析其在增生性瘢痕(Hypertrophic scar,HS)患者中对成纤维细胞增殖与迁移的影响及作用机制。方法:通过circRNA序列和定量聚合酶链反应(PCR技术)检测正常皮肤成纤维细胞(NSFBs)和增生性瘢痕患者成纤维细胞(HSFBs)中circ-0030042的表达。用CCK8检测法检测转染48 h后的HSFBs细胞增殖情况。利用stubRFP-sensGFP-LC3基因转染、流式细胞仪及电子显微镜观察circ-0030042对miR-145/PTEN轴调控VEGF水平的表达。利用生物信息学分析、RNA免疫沉淀、免疫荧光检测等方法,揭示circ-0030042介导HS患者成纤维细胞增殖与迁移的作用机制。结果:circ-0030042在增生性瘢痕中显著上调,过表达时作为VEGF海绵抑制miR-145诱导的成纤维细胞,维持体内稳定性。此外,circ-0030042通过海绵化VEGF水平并阻断其miR-145捕获转录因子(FOXO1)mRNA来影响自噬,而circ-0030042诱导FOXO1的抑制被VEGF水平过表达或circ-0030042结合减少所抵消。过表达circ-0030042对成纤维细胞的增殖抑制与VEGF表达的抑制作用被过表达miR-145部分抵消。结论:干扰circ-0030042通过靶向下调miR-145/PTEN轴进而抑制HSFBs细胞的增殖与迁移,进一步诱导恶性细胞凋亡。
