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  • Title: [The expression of SnoN in human hypertrophic scar fibroblasts and the mechanism of its participation in hypertrophic scar formation].
    Author: Kuang F, Zhang Z, Chen B, Liu CL, Zhao YY, Xu ZR, Li XJ.
    Journal: Zhonghua Shao Shang Za Zhi; 2017 Oct 20; 33(10):634-638. PubMed ID: 29056026.
    Abstract:
    Objective: To investigate the expression of SnoN in human hypertrophic scar fibroblasts and the mechanism of its participation in hypertrophic scar formation. Methods: Eight patients with hypertrophic scar after burn in need of surgery were admitted in our unit from January to October 2013, and then hypertrophic scar tissue and normal skin tissue of full-thickness skin donor site resected by surgery of the patients were collected. Hypertrophic scar fibroblasts and normal skin fibroblasts of patients were isolated with method of explant culture and then sub-cultured. Cells of the third to fifth passage were used in the following experiments. (1) The protein expressions of SnoN of hypertrophic scar fibroblasts and normal skin fibroblasts were assessed with Western blotting. (2) The mRNA expressions of SnoN of another batch of hypertrophic scar fibroblasts and normal skin fibroblasts were determined with reverse transcription polymerase chain reaction. (3) Another batch of hypertrophic scar fibroblasts and normal skin fibroblasts were treated with 10 ng/mL transforming growth factor beta1 (TGF-β(1)) for 30 min, 1 h, 2 h, and 6 h, respectively, and then the protein expressions and mRNA expressions of SnoN of untreated cells and treated cells were detected as above. Data were processed with one way analysis of variance and independent sample t test. Results: (1) The protein expression of SnoN of hypertrophic scar fibroblasts was 0.020±0.003, significantly lower than that of normal skin fibroblasts (0.032±0.005, t=7.19, P<0.05). (2) The mRNA expression of SnoN of hypertrophic scar fibroblasts was 0.407±0.157, with no significant difference from that of normal skin fibroblasts (0.339±0.095, t=-1.29, P>0.05). (3) The protein expression of SnoN of normal skin fibroblasts was increased in a time-dependent fashion with the TGF-β(1) stimulation, and the protein expressions of SnoN of cells treated with TGF-β(1) for 30 min, 1 h, 2 h, and 6 h were significantly higher than those of untreated cells (with t values from 2.27 to 27.89, P values below 0.05). The protein expression of SnoN of hypertrophic scar fibroblasts was decreased in a time-dependent fashion with the TGF-β(1) stimulation, and the protein expressions of SnoN of cells treated with TGF-β(1) for 30 min, 1 h, 2 h, and 6 h were obviously lower than those of untreated cells (with t values from 10.80 to 13.85, P values below 0.05). (4) The mRNA expressions of SnoN of normal skin fibroblasts and hypertrophic scar fibroblasts were both increased in a time-dependent fashion with the TGF-β(1) stimulation, and the mRNA expressions of SnoN of the two types of cells treated with TGF-β(1) for 30 min, 1 h, 2 h, and 6 h were both significantly higher than those of untreated cells (with t values from 18.16 to 58.22, P values below 0.05). Conclusions: The protein expression of SnoN in hypertrophic scar fibroblasts is reduced, which weakens its inhibitory effect on TGF-β(1) signal, thus amplifying the TGF-β(1) signal, and it may participate in the formation of hypertrophic scar. 目的： 探讨人增生性瘢痕Fb中SnoN的表达及其参与增生性瘢痕形成的机制。 方法： 收集笔者单位2013年1—10月收治的8例烧伤后瘢痕增生需行手术治疗患者的增生性瘢痕组织及其手术切除的全层皮肤供皮区的正常皮肤组织。组织块培养法分离人增生性瘢痕Fb和正常皮肤Fb，并进行传代培养，取第3～5代细胞进行以下实验。(1)取增生性瘢痕Fb和正常皮肤Fb，蛋白质印迹法检测2种细胞中SnoN的蛋白表达。(2)另取增生性瘢痕Fb和正常皮肤Fb，RT－PCR法检测2种细胞中SnoN mRNA表达。(3)另取增生性瘢痕Fb和正常皮肤Fb，加入10 ng/mL TGF－β(1)刺激30 min和1、2、6 h后，同前检测未刺激及刺激后2种细胞中SnoN的蛋白及mRNA表达。对数据行单因素方差分析、独立样本t检验。 结果： (1)增生性瘢痕Fb中SnoN的蛋白表达量为0.020±0.003，明显低于正常皮肤Fb的0.032±0.005(t＝7.19，P<0.05)。(2)增生性瘢痕Fb中SnoN mRNA表达量为0.407±0.157，与正常皮肤Fb的0.339±0.095无明显差异(t＝－1.29，P>0.05)。(3)正常皮肤Fb中SnoN蛋白表达量在TGF－β(1)刺激下呈时间依赖性增高，刺激30 min和1、2、6 h，细胞中SnoN蛋白表达量均明显高于未刺激细胞(t值为2.27～27.89，P值均小于0.05)。增生性瘢痕Fb中SnoN蛋白表达量在TGF－β(1)刺激下呈时间依赖性降低，刺激30 min和1、2、6 h，细胞中SnoN蛋白表达量均明显低于未刺激细胞(t值为10.80～13.85，P值均小于0.05)。(4)正常皮肤Fb和增生性瘢痕Fb中SnoN mRNA表达量在TGF－β(1)刺激下均呈时间依赖性增高，2种细胞刺激30 min和1、2、6 h，其SnoN mRNA表达量均明显高于未刺激细胞(t值为18.16～58.22，P值均小于0.05)。 结论： 增生性瘢痕Fb中SnoN蛋白表达减少，导致其对TGF－β(1)信号抑制作用减弱，从而放大了TGF－β(1)信号，可能参与了增生性瘢痕的形成。.
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