柔软纤维蛋白水凝胶对活化态肝星状细胞转分化行为的影响

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白水,凝胶,活化,分化,柔软
柔软纤维蛋白水凝胶对活化态肝星状细胞转分化行为的影响

陈烨;胡文慧;曾柱;李兴;蒋天鹏;王黎洲;周石;宋杰

【期刊名称】《中山大学学报（医学科学版）》【年(卷),期】2018(039)001

【摘要】[Objective]To determine whether activated hepatic stellate cell(HSC)become quiescent phenotype if cultured in a soft extracellular matrix(ECM).[Methods]HSC-T6 cells which stably expressed

myofibroblast(MFB) phenotype,were cultured in the 2D and 3D in-vitro matrix culture models that were constructed by 1 mg/mL fibrin gel. The proliferation activity of HSC-T6 was determined by cell counting kit-8(CCK8)assay at different time points(24 h, 48 h,72 h,96 h).

Immunofluorescence and gelatin zymography analysis were performed to detect the expression of α-SMA,MMP-2,and MMP-9. Moreover,the cellular surface morphology and deformability was detected by AFM.[Results]After cultured in the fibrin gel,the cellular proliferation activity,expression of α-SMA,MMP-2 and MMP-9,and cellular deformability of activated HSC-T6 was obviously down-regulated in contrast to the control group (P<0.05),and there were also differences in the inhibition of cellular proliferation activity,the assembly of α-SMA stress fiber and the effect of cellular deformability between 2D and 3D fibrin gel cultured(P<0.05).[Conclusions]Soft fibrin gel inhibited the

transdifferentiation of activated HSC-T6,but could not make it revert to α-

SMA negative quies-cent phenotype.%[目的]探讨活化态肝星状细胞(HSC)在柔软细胞外基质(ECM)中重新恢复为静息态的可能性.[方法]稳定表达肌成纤维细胞(MFB)表型的HSC-T6培养于由1 mg/mL纤维蛋白水凝胶构建的二维(2D)或三维(3D)体外培养基质模型之中,利用细胞活力检测试剂盒(CCK8)测定各时间段(24 h、48 h、72 h、96 h)的细胞增殖活力,免疫荧光及明胶酶谱法检测细胞α平滑肌肌动蛋白(α-SMA)、基质金属蛋白酶-2(MMP-2)及MMP-9的表达,原子力显微镜(AFM)探测细胞表面形貌及其变形能力.[结果]经纤维蛋白水凝胶培养后,活化态HSC-T6的细胞增殖活力、α-SMA、MMP-2和MMP-9的表达以及细胞变形能力均发生明显下调(P<0.05),而2D组纤维蛋白水凝胶对细胞增殖活力的抑制、α-SMA应力纤维的形成以及细胞变形能力的影响与3D组对比亦存在不同程度的差异(P<0.05).[结论]柔软的纤维蛋白水凝胶可抑制活化态HSC转分化,但无法使其逆转为α-SMA阴性的静息态表型. 【总页数】9页(P9-17)

【作者】陈烨;胡文慧;曾柱;李兴;蒋天鹏;王黎洲;周石;宋杰

【作者单位】贵州医科大学医学影像学院,贵州贵阳550004;贵州医科大学生物与工程学院,贵州贵阳550004;贵州医科大学生物与工程学院,贵州贵阳550004;贵州医科大学医学影像学院,贵州贵阳550004;贵州医科大学医学影像学院,贵州贵阳550004;贵州医科大学医学影像学院,贵州贵阳550004;贵州医科大学医学影像学院,贵州贵阳550004;贵州医科大学医学影像学院,贵州贵阳550004 【正文语种】中文【中图分类】R575 【相关文献】

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