华山参通过TGF-β1/Smad3通路对体外纤维化的调节作用  

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

作　　者：商丹丹 周鸿 黄蔓如 江永萍 刘彩艳 刘锐 张思祺 于海洋[1] 王涛[1] SHANG Dandan;ZHOU Hong;HUANG Manru;JIANG Yongping;LIU Caiyan;LIU Rui;ZHANG Siqi;YU Haiyang;WANG Tao(Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;The Sixth Chinese Medicine Factory of Tianjin Darentang Group Co.,Tianjin 300401,China;Phytochemical Research Department,Research Institute of Traditional Chinese Medicine,Tianjin 300457,China)

机构地区：[1]天津中医药大学,天津301617 [2]津药达仁堂集团股份有限公司第六中药厂,天津300401 [3]津药达仁堂集团股份有限公司中药研究院植化研究室,天津300457

出　　处：《天津中医药大学学报》2024年第6期523-530,共8页Journal of Tianjin University of Traditional Chinese Medicine

摘　　要：[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。[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,β-galactosidase 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 TGF-β1/Smad3 pathway.

关 键 词：华山参 肺纤维化 TGF-β1/Smad3 

分 类 号：R563[医药卫生—呼吸系统]