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目的:研究补益营卫方对衰老人皮肤成纤维细胞TGFβ1/Smads信号蛋白基因表达的影响。方法:通过传代培养复制衰老人皮肤成纤维细胞模型,利用荧光定量PCR法检测补益营卫方对衰老人皮肤成纤维细胞TGFβ1/Smads信号蛋白基因表达。结果:人皮肤成纤维细胞衰老时Smad2基因的表达水平,与年轻细胞相比,变化趋势基本一致。补益营卫药物处理后Smad2基因的表达,24h和72h时,分别是衰老人皮肤成纤维细胞基因相对表达量的2.85倍和2.5倍;而48 h时则两者相当。衰老时Smad3基因的表达水平,24 h和48 h时,衰老人皮肤成纤维细胞比年轻细胞分别下降了29%和36%;72 h则增加了3.95倍。补益营卫药物处理后Smad3基因的表达,24 h、48 h和72 h时,分别是衰老人皮肤成纤维细胞的3.15倍、2.6倍和1.6倍。衰老时Smad4基因的表达水平,24 h时比年轻细胞下降了32%,但是,48 h和72 h时则分别是年轻人皮肤成纤维细胞的1.6倍和约3倍。补益营卫药物处理后Smad4基因的表达24 h、48 h和72 h分别是衰老组的2.8倍、1.6倍和2倍。衰老时Smad7基因的表达水平,24 h和48 h时分别比年轻细胞下降了88%和64%。但是,72 h时则达到年轻人皮肤成纤维细胞表达水平的1.9倍。补益营卫药物处理后Smad7基因的表达,其24h时是衰老人皮肤成纤维细胞表达水平的4.24倍,但是,48 h和72 h则分别下降了24%和33%。结论:衰老时TGFβ1/Smads信号蛋白表达紊乱;补益营卫方对衰老人皮肤成纤维细胞Smad2、Smad3、Smad4基因表达有促进作用;对Smad7基因表达表现为双向调节作用。
Objective: To study the effect of BuYiYangWeiFang on gene expression of TGFβ1 / Smads signaling in human skin fibroblasts. Methods: The human dermal fibroblasts were subcultured and subcultured, and the expression of TGFβ1 / Smads signaling genes in human skin fibroblasts was detected by real-time fluorescence quantitative PCR. Results: The expression level of Smad2 in human skin fibroblasts was basically the same as that in young cells. The expression of Smad2 gene in drug-ying-wei drug-treated group was 2.85-fold and 2.5-fold higher than that in the senescent human skin fibroblasts at 24 and 72 hours, respectively. At 24 and 48 h, the expression of Smad3 in aging cells decreased by 29% and 36% respectively compared with that in young cells, and increased by 3.95 times in 72 hours. The expression of Smad3 gene after drug treatment of BuYiYingWei was 3.15 times, 2.6 times and 1.6 times higher than that of aging human skin fibroblasts at 24 h, 48 h and 72 h, respectively. The level of Smad4 expression in aging cells was 32% lower than that of young cells at 24 hours, but 1.6 times and about 3 times higher than that of young human skin fibroblasts at 48 and 72 hours, respectively. The expression of Smad4 gene in the treatment of BuYiYingWei drug was 2.8 times, 1.6 times and 2 times of the aging group at 24 h, 48 h and 72 h, respectively. The expression of Smad7 in aging cells decreased by 88% and 64% respectively compared with young cells at 24 h and 48 h. However, at 72 h, it reached 1.9 times the level of young human dermal fibroblasts. The expression of Smad7 gene after treatment of BuYiYingWei was 4.24 times of the expression of human dermal fibroblasts at 24 hours, but decreased by 24% and 33% at 48 and 72 h, respectively. CONCLUSION: TGFβ1 / Smads signaling protein expression is disturbed during aging. BuYiYangWeiFang can promote Smad2, Smad3 and Smad4 gene expression in aging human dermal fibroblasts and play a bidirectional role in Smad7 gene expression.