硅橡胶硬度通过自噬调控成纤维细胞增殖及α- SMA、CollagenⅠ的表达

Silicone rubber hardness regulates fibroblasts proliferation and expression of α-SMA and CollagenⅠ by autophagy

目的探讨不同硬度硅橡胶对人真皮成纤维细胞增殖及其表达α-SMA和CollagenⅠ的影响,并研究自噬在其中所起的作用。方法将A、B组分液体硅胶按不同比例混合固化制成一系列不同硬度的硅橡胶片,测定其邵氏硬度、断裂伸长率、水接触角、表面形貌等机械性能。利用CCK-8检测成纤维细胞在不同硬度硅橡胶表面的增殖情况,而后选取细胞增殖存在显著差异的两种硬度硅橡胶进一步实验,采用Western blot和ELISA法分别检测正常条件下、施加自噬抑制剂3-MA和诱导剂西罗莫司后成纤维细胞自噬标志蛋白和α-SMA、CollagenⅠ的表达情况。结果成纤维细胞增殖能力随硅橡胶硬度的增加而增加,差异有统计学意义(P<0.05)。高硬度硅橡胶上的成纤维细胞具有更高的自噬活性,且α-SMA、Collagen I的表达量比低硬度硅橡胶更低,差异有统计学意义(P<0.01)。结论硅橡胶硬度可通过自噬调控成纤维细胞的增殖,高硬度硅橡胶上调成纤维细胞自噬活性从而促进成纤维细胞增殖并抑制其表达α-SMA和CollagenⅠ。

Objective To investigate the effect of different hardness of silicone rubber(SRs)on the proliferation and expression ofα-SMA and CollagenⅠin human dermal fibroblasts(HFBs),and study the role of autophagy in the process.Methods In this study,we prepared a series of SRs with different hardness by changing the ratio of liquid silica gel component A to B.The mechanical properties of SRs such as A-shore hardness,breaking elongation,water contact angle and surface morphology were measured.CCK-8 assay was used to study the proliferation of HFBs cultivated on the surface of these SRs.Then,2 kinds of SRs with significant differences in hardness and cell proliferation were subjected,and Western blotting and ELISA were employed to detect the expression of autophagy markers,α-SMA and CollagenⅠin the HFBs after the treatment with autophagy inhibitor 3-MA and inducer rapamycin respectively.Results The proliferation of HFBs was obviously increased with the increase of SRs hardness(P<0.05).Furthermore,the HFBs cultured on harder SRs surface had higher autophagy activity,and lower expression levels ofα-SMA and CollagenⅠwhen compared with the cells on softer SRs surface(P<0.01).Conclusion The SRs hardness may affect the proliferation of HFBs by regulating autophagy.High hardness SRs promotes the proliferation of HFBs and inhibits the expression ofα-SMA and CollagenⅠby enhancing autophagy activity.