当归红芪超滤物对辐射后心肌成纤维细胞的增殖和胶原分泌的影响

Effect of Ultra-filtration Extraction from Angelicae Sinensis Radix and Hedysari Radix on Cell Proliferation and Collagen Secretion of Myocardial Fibroblasts Induced by Radiation

目的：研究当归红芪超滤物对辐射后心肌成纤维细胞增殖和胶原分泌的影响。方法：采用差速贴壁法原代分离培养大鼠心肌成纤维细胞;当归红芪超滤物干预或射线辐射心肌成纤维细胞后,采用噻唑蓝（MTT）比色法检测细胞的增殖,酶联免疫吸附法（ELISA）检测细胞分泌Ⅰ型胶原（COL-Ⅰ）的含量;心肌成纤维细胞通过2＇,7＇-二氯荧光黄双乙酸盐（DCFH-DA）染色在流式细胞仪下检测活性氧（ROS）的含量;采用实时荧光定量聚合酶链式反应法（Real-time PCR）检测心肌成纤维细胞中COL-Ⅰ和转化生长因子-β（TGF-β）的mRNA表达水平;蛋白免疫印迹法（Western blot）检测相关蛋白的表达。结果：低剂量射线（0.5 Gy）辐射心肌成纤维细胞后,细胞内ROS的升高促进细胞增殖和胶原分泌的作用;当归红芪超滤物可以促进细胞中转录因子E2相关因子2（Nrf2）蛋白的核转位来清除ROS,最终抑制辐射诱导的心肌成纤维细胞的增殖和胶原分泌;并且当归红芪超滤物明显抑制心肌成纤维细胞中COL-Ⅰ和TGF-βmRNA表达。结论：当归红芪超滤物可以抑制低剂量辐射诱导的心肌成纤维细胞增殖和胶原分泌。

Objective： To investigate the effect of ultra-filtration extraction from Angelicae Sinensis Radix and Hedysari Radix on the cell proliferation and collagen deposition of myocardial fibroblasts induced by radiation.Method： Rat myocardial fibroblasts were separated and cultured by cell adhesion. The cell proliferation was determined by thiazolyl blue tetrazolium bromide（ MTT） method,and the collagen-Ⅰ（ COL-Ⅰ） of secretion was determined by enzyme-linked immuno sorbent assay（ ELISA） method after ultra-filtration extraction from Angelicae Sinensis Radix and Hedysari Radix or radiation to myocardial fibroblasts. The changes of reactive oxygen species（ ROS） were determined by flow cytometry with indicator 2＇,7＇-dichloride fluorescent yellow double acetate（ DCFH-DA） staining. The mRNA expressions of COL-Ⅰ and transforming growth factor-beta（ TGF-β） were determined by Real-time polymerase chain reaction（ PCR）, and the expressions of relative proteins were determined by Western blot. Result： The rising cellular ROS promoted cell proliferation and collagen deposition of myocardial fibroblasts induced by low-dose radiation（ 0. 5 Gy）,which however was inhibited through ultra-filtration extraction from Angelicae Sinensis Radix and Hedysari Radix by promoting nuclear translocation of nuclear factor E2 related factor 2（ Nrf2） and eliminating ROS. The mRNA expressions of COL-Ⅰ and TGF-β were both inhibited by ultra-filtration extraction from Angelicae Sinensis Radix and Hedysari Radix. Conclusion： Ultra-filtration extraction from Angelicae Sinensis Radix and Hedysari Radix could inhibit the cell proliferation and collagen deposition of myocardial fibroblasts induced by low-dose radiation.