PTEN介导紫草素抑制人增生性瘢痕成纤维细胞的胶原沉积

Shikonin inhibits collagen deposition of human hypertrophic scar fibroblasts via PTEN

目的从PTEN角度探讨紫草素对人增生性瘢痕成纤维细胞胶原沉积的影响。方法苏木精-伊红染色(HE染色)和马松染色(Masson染色)鉴定增生性瘢痕组织,免疫荧光鉴定人增生性瘢痕成纤维细胞(HSFBs);Western blot检测人增生性瘢痕(HS)和同一个体正常皮肤(NS)组织中PTEN表达水平;将HSFBs随机分组为:DMSO组、shikonin组、control组、si-NC组、si-PTEN组、shikonin+si-NC组和shikonin+si-PTEN组;Western blot检测各组中PTEN和胶原相关蛋白(Col1、Col3、α-SMA)的表达。结果与正常皮肤组织相比,PTEN在增生性瘢痕组织中呈低表达(P<0.05);用IC50浓度(13.46μmol/L)的紫草素干预人增生性瘢痕成纤维细胞24 h后,人增生性瘢痕成纤维细胞中Col1、Col3和α-SMA的表达降低(P<0.05),而PTEN的表达升高(P<0.05);转染si-PTEN后,与si-NC组相比,人增生性瘢痕成纤维细胞中Col1,Col3和α-SMA的表达升高(P<0.05);共转染shikonin+si-PTEN后,与shikonin+si-NC组相比,人增生性瘢痕成纤维细胞中Col1、Col3和α-SMA的表达升高(P<0.05)。结论紫草素可通过上调PTEN的表达抑制增生性瘢痕成纤维细胞的胶原沉积。

Objective To investigate the effect of shikonin on the collagen deposition of human hypertro-phic scar fibroblasts from the perspective of PTEN.Methods Hematoxylin-eosin staining and Masson staining were used to identify hypertrophic scar tissue.Immunofluorescence was used to identify fibroblasts.The relative expression level of PTEN was detected by Western blot at the tissue level.Hypertrophic scar fibroblasts were randomly divided into dimethyl sulfone group,shikonin group,control group,si-NC group,shikonin+si-NC group and shikonin+si-PTENC group,western blot were used to detect the expression of PTEN and collagen-related proteins(Col1,Col3 andα-SMA)at protein levels,respectively.Results Compared with normal skin,PTEN was lower expressed in hypertrophic scar(P<0.05).shikonin was dissolved in dimethyl sulfone to make drug solution at IC50 concentration(13.46μmol/L),which was used for human hypertrophic scar fibroblasts cell culture for 24 hours,shikonin down-regulated the expression levels of Col1,Col3 andα-SMA in human hypertrophic scar fibroblasts,while up-regulated the expression of PTEN(P<0.05).Transfected with si-PTEN,the expressions of Col1,Col3 andα-SMA in human hypertrophic scar fibroblasts were increased compared with the si-NC group(P<0.05).After co-transfection with shikonin+si-PTEN,the expressions of Col1,Col3 andα-SMA in human hypertrophic scar fibroblasts were increased compared with shikonin+si-NC group(P<0.05).Conclusion Shikonin can inhibit collagen deposition in hypertrophic scar fibroblasts by up-regulating PTEN expression.