华山参通过TGF-β1/Smad3通路对体外纤维化的调节作用

[目的] 研究华山参对重组小鼠转化生长因子 β1 以及香烟提取物（CSE）对小鼠胚胎成纤维细胞（NIH-3T3）和人源支气管上皮细胞（BEAS-2B）的影响及相关机制。[方法] 噻唑蓝（MTT）法筛选华山参生物碱、非生物碱、总提取物对 NIH-3T3 细胞的安全浓度；重组小鼠转化生长因子 β1 诱导 NIH-3T3 细胞 24 h，进行天狼猩红染色，并计算相对胶原沉积率；MTT 法分别筛选 7 种华山参生物碱单体对 BEAS-2B 细胞的安全浓度；CSE 处理 BEAS-2B细胞 24 h 后，进行 β-半乳糖苷酶（β-Gal）染色；蛋白免疫印迹法（Western Blot）检测 TGF-β1、Smad3、NLRP3 蛋白的表达水平。[结果] 华山参生物碱、非生物碱、总提取物均能抑制重组小鼠转化生长因子 β1 诱导的 NIH-3T3 细胞发生胶原沉积（P 均<0.05）；东莨菪碱和去水阿托品可以显著抑制 CSE 诱导的 BEAS-2B 细胞发生衰老（P 均<0.05），并且能够显著抑制 TGF-β1、Smad3、NLRP3 蛋白的表达（P 均<0.05）。[结论] 华山参可能通过抑制 TGF-β1/Smad3 通路来调节体外肺纤维化的进程。

Study on the in vitro mechanism of Physochlaina infundibularis Kuang in intervening pulmonary fibrosis

[Objective] To study the effects of Physochlaina infundibularis Kuang on Recombinant Mouse TGF-β1 （TGF-β1） as well as cigarette smoke extract（CSE） on mouse embryonic fibroblasts（NIH-3T3） and human bronchial epithelial cells（BEAS-2B） and related mechanisms. [Methods] MTT method was used to screen the safe concentrations of Physochlaina infundibularis Kuang alkaloids，non-alkaloids，and total extracts on NIH-3T3 cells；NIH-3T3 cells were induced by TGF-β1 for 24 h，stained with Sirius Scarlet，and the relative collagen deposition rate was calculated；MTT method was used to screen the safe concentrations of 7 Physochlaina infundibularis Kuang alkaloidal monomers on BEAS-2B cells，respectively；BEAS-2B cells were treated with CSE After 24，βgalactosidase staining was performed；Western blot method was used to detect the expression levels of TGF-β1， Smad3 and NLRP3 proteins. [Results] Physochlaina infundibularis Kuang alkaloids，non-alkaloids，and total extracts significantly inhibited TGF-β1-induced collagen deposition in NIH-3T3 cells（all P<0.05）；scopolamine and dehydro-atropine significantly inhibited CSE-induced senescence in BEAS-2B cells （all P<0.05），and were able to significantly inhibit the expression of TGF-β1，Smad3，and NLRP3 proteins （all P <0.05）. [Conclusion] Physochlaina infundibularis Kuang may regulate the progression of pulmonary fibrosis in vitro by inhibiting the TGFβ1/Smad3 pathway.