论文部分内容阅读
目的:探讨黄芪总苷液对人瘢痕疙瘩成纤维细胞的抑制作用。方法:以正常皮肤为对照,用免疫组化检测瘢痕疙瘩组织标本中转化生长因子-β(transforming growth factor-β,TGF-β)及其转导因子(SMAD family member,Smad)的表达情况;不同浓度黄芪总苷液干预HFF-1人皮肤成纤维细胞后,MTT法检测其最适浓度;Real time-PCR测定成纤维细胞TGF-β的转导因子Smad2(SMAD family member 2),Smad3(SMAD family member 3),Smad4(SMAD family member 4),Smad7(SMAD family member 7)mRNA的表达;Western blot检测成纤维细胞TGF-βRⅡ,Smad2,Smad3,Smad4,Smad7的蛋白水平表达。结果:与正常皮肤组织相比,瘢痕疙瘩组织Smad蛋白表达水平明显提高(P<0.05),TGF-βRⅡ蛋白的表达水平在两组中没有明显差异(P>0.05)。MTT检测最适给药浓度为0.5μg·ml-1。两组细胞TGF-β/Smad信号通路相关蛋白表达量比较,给药组Smad2的表达水平明显提高,Smad3的表达水平明显降低(P<0.05)。结论:黄芪总苷液可能通过过表达Smad2以及抑制Smad3的表达来影响TGF-β/Smad通路过程从而抑制成纤维细胞的形成。
Objective: To investigate the inhibitory effect of astragalus total glycosides on human keloid fibroblasts. Methods: The normal skin was used as control. The expression of transforming growth factor-β (TGF-β) and its transcription factor (SMAD) in keloid tissue specimens were detected by immunohistochemistry. The optimal concentration of astragalus total glycosides solution in HFF-1 human dermal fibroblasts was determined by MTT assay. The expressions of Smad2 (SMAD family member 2), Smad3 The expression of TGF-βRⅡ, Smad2, Smad3, Smad4 and Smad7 in fibroblasts were detected by Western blot. The expression of Smad4 (SMAD family member 4) and Smad7 (SMAD family member 7) mRNA was detected by Western blot. Results: Compared with normal skin tissue, the expression of Smad protein in keloid tissue was significantly increased (P <0.05). The expression of TGF-βRⅡ protein in the two groups showed no significant difference (P> 0.05). The optimal concentration of MTT assay was 0.5μg · ml-1. Compared with the expression of TGF-β / Smad signaling pathway-related proteins in both groups, Smad2 expression and Smad3 expression were significantly decreased (P <0.05). Conclusion: Astragalosides solution may inhibit the formation of fibroblasts by overexpression of Smad2 and inhibition of Smad3 expression by affecting the TGF-β / Smad pathway.