饥饿诱导增生性瘢痕成纤维细胞自噬发生

 ［摘要］目的:探讨饥饿诱导的增生性瘢痕成纤维细胞自噬的发生。方法:原代培养增生性瘢痕成纤维细胞至对数生长期，用EBSS（Earle’s balanced salt solution）代替培养液，分别饥饿0 h、1 h、2 h、3 h，应用Western blotting和定量PCR(qRT-PCR)方法分别检测成纤维细胞微管相关蛋白1轻链3（microtubule-associated protein 1 light chain 3, LC-3）和自噬相关蛋白Beclin-1的蛋白和mRNA表达，单丹磺酰戊二胺(monodansylcadaverine,MDC)染色和透射电镜观察自噬体。结果:Western blotting和qRT-PCR 检测LC3和Beclin-1 蛋白和mRNA表达在饥饿1 h开始增加，2 h达到高峰，3 h逐渐下降，均明显大于对照组（P<0.05）。荧光显微镜和电镜下观察到饥饿2 h诱导的成纤维细胞出现自噬囊泡。结论: 饥饿可诱导增生性瘢痕成纤维细胞发生自噬,可能与增生性瘢痕的形成有关。

Starvation induces autophagy of hypertrophic scar fibroblasts

［ABSTRACT］AIM: To investigate the starvation-induced autophagy of  hypertrophic scar fibroblasts. METHODS: Primary human fibroblasts from hypertrophic scars were isolated and the fibroblasts in logarithmic growth phase were cultured with amino acid-free Earle's balanced salt solution (EBSS) instead of the DMEM medium. The cells were collected at the time points of 0 h, 1 h, 2 h and 3 h after EBSS culture. The expression of microtubule-associated protein 1 light chain 3 (LC-3) and autophagy-related protein Beclin-1 was detected by Western blotting and qRT-PCR. Autopagosomes in fibroblasts were observed by electron microscopy and monodansylcadaverine (MDC) staining. RESULTS: The expression of LC3 and Beclin-1 increased at 1 h after starvation, reached to the highest level at 2 h after starvation, and then began to decline, which were still higher than that in control group. Compared with control group, the autopagosomes were observed in the fibroblasts under fluorescence microscope and election microscope at 2 h after starvation. CONCLUSION: Autophagy in fibroblasts of hypertrophic scars can be induced by starvation, which may be related to the formation of hypertrophic scars.