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目的观察中药天龙喘咳灵对转化生长因子-β1(TGF-β1)诱导的气道上皮下成纤维细胞(HPBFs)α-平滑肌肌动蛋白(α-SMA)表达的影响,探讨其可能的信号转导机制。方法2005—2006年,在广州市红十字会医院呼吸科于原代培养的HPBFs中,分别加入天龙喘咳灵合剂、天龙喘咳灵主要组分(淡附子、青天葵和法夏)的单味药预孵育2h,再加入TGF-β1(10μg/L)共孵育;刺激10min和30min,分别提取细胞总蛋白,Western免疫印迹方法检测磷酸化p38MAPK、ERK1/2和smad2的蛋白表达;刺激20h和72h,提取细胞总RNA及蛋白,检测α-SMA蛋白及mRNA的表达。结果天龙喘咳灵组和淡附子组α-SMA蛋白和mRNA的表达均较单纯TGF-β1刺激组减少(P<0·05),天龙喘咳灵组对α-SMA蛋白和mRNA表达量的抑制程度高于淡附子组;天龙喘咳灵组和淡附子组磷酸化ERK1/2的表达明显低于单纯TGF-β1刺激组,而青天葵和法夏两组与单纯TGF-β1组相似。各药物处理组对TGF-β1诱导的磷酸化p38和smad2的表达无明显影响。结论天龙喘咳灵抑制HPBFs的α-SMA表达,可能是通过降低TGF-β1诱导的ERK1/2磷酸化水平实现的,并且其中的组分淡附子可能起主要作用。
Objective To observe the effect of traditional Chinese medicine Tianlongchuankeji on the expression of α-smooth muscle actin (α-SMA) induced by transforming growth factor-β1 (TGF-β1) in airway subepithelial fibroblasts (HPBFs) and to explore the possible signal transduction. Leading mechanism. Methods From 2005 to 2006, in the primary culture of HPBFs in the Department of Respiratory Medicine of the Guangzhou Red Cross Hospital, Tianlongchuankekeling Mixture and the main components of Tianlongchuankeji Spirit (Tanfuzi, Qingtiankui and Faxia) were added separately. Pre-incubation with flavoring drugs for 2 h, and then adding TGF-β1 (10 μg/L) were co-incubated; stimulated for 10 min and 30 min, respectively, extracted total cellular protein, and detected the expression of p38MAPK, ERK1/2, and smad2 by Western blotting; stimulated for 20 h At 72h, total cellular RNA and protein were extracted to detect the expression of α-SMA protein and mRNA. Results The expression of α-SMA protein and mRNA in the Tianlongchuankeling group and the light aconite group was lower than that of the TGF-β1 alone group (P<0.05). The expression of α-SMA protein and mRNA in the Tianlongchuankekeling group was lower than that in the TGF-β1 group alone. The degree of inhibition was higher than that of the light aconite group; the expression of phosphorylated ERK1/2 in the Tianlongchuankeling group and the light aconite group was significantly lower than that of the TGF-β1 alone group, while the two groups were similar to the TGF-β1 group. Each drug treatment group had no significant effect on TGF-β1-induced phosphorylation of p38 and smad2. Conclusion Tianlongchuankeling inhibits the expression of α-SMA in HPBFs, which may be achieved by decreasing the level of ERK1/2 phosphorylation induced by TGF-β1. Among these components, light aconite aconite may play a major role.