中文摘要：

目的 探讨转化生长因子（TGF-β1）是否可通过活性氧（ROS）激活c-Jun末端激酶（JNK）通路,刺激肺成纤维细胞增殖和Ⅰ/Ⅲ型胶原合成增加,促进矽肺纤维化形成.方法 在检测TGF-β1对肺成纤维细胞作用的实验中,肺成纤维细胞分为对照组（0.4％血清）和TGF-β1组（5μg/L）.在检测抗氧化剂NAC（N-乙酰-L-半胱氨酸）是否抑制了TGF-β1对肺成纤维细胞作用的实验中,肺成纤维细胞分为对照组（0.4％血清）、H2O2组（0.1mmol/L,阳性对照）、H2O2＋NAC组（10 mmol/L）、TGF-β1组（5μg/L）、TGF-β1＋NAC组.利用MTT法测定细胞增殖,蛋白免疫印迹法（western blot）测定Ⅰ/Ⅲ型胶原及JNK表达,免疫荧光法检测8-羟化脱氧鸟苷（8-OHdG,一种DNA氧化产物）水平.结果 在检测TGF-β1对肺成纤维细胞作用的实验中,与对照组比较,TGF-β1组肺成纤维细胞增殖增强、Ⅰ/Ⅲ型胶原、JNK磷酸化及8-OHdG水平增高,差异有统计学意义（P＜0.05）.在检测NAC是否抑制了TGF-β1对肺成纤维细胞作用的实验中,与对照组比较,H2O2组和TGF-β1组细胞增殖的吸光度值,Ⅰ/Ⅲ型胶原及JNR磷酸化、8-OHdG水平均增高,但与H2O2组和TGF-β1组比较,H2O2＋NAC组和TGF-β1＋NAC组的吸光度值,Ⅰ/Ⅲ型胶原及JNR磷酸化、8-OHdG水平均表达下降,差异有统计学意义（P＜0.05）.结论 TGF-β1可诱导肺成纤维细胞生成ROS,并由此促进JNK磷酸化,最终引起肺成纤维细胞增殖和Ⅰ/Ⅲ型胶原合成的增多,减少ROS,可有效抑制TGF-β1的作用.

英文摘要：

Objective This study will explore whether reactive oxygen species （ROS） is involved in TGF-β1-induced JNK activation,pulmonary fibroblast proliferation and collagen type Ⅰ and Ⅲ synthesis.Methods Pulmonary fibroblasts were randomly divided into control （0.4% serum） and TGF-β1 （5 μg/L）groups to detect whether TGF-β1 could induce pulmonary fibroblast proliferation,synthesis of collagen Ⅰ and Ⅲ,phosphorylated-JNK （p-JNK） and 8-OHdG （indicator of ROS）; while in the part to explore whether NAC （N-acetyl-L-cysteine,antioxidants） has the inhibitory role in TGF-β1-induced pulmonary fibroblast,it did control （0.4% serum）,H2O2 （0.1 mmol/L,positive control）,H2O2＋NAC （10 mmol/L）,TGF-β1 （5 μg/L）,TGF-β1＋NAC groups.Pulmonary fibroblast proliferation,8-OHdG levels,expressions of JNK and collagen Ⅰ and Ⅲ were used by MTT assay,immunofluorescence and western blot respectively.Results In the experiments to detect the effect of TGF-β1 on pulmonary fibroblasts,compared with control,TGF-β1 significantly stimulated pulmonary fibroblast proliferation and increased collagen Ⅰ and Ⅲ protein,p-JNK and 8-OHdG levels.In the next experiments to explore whether NAC has the inhibitory role in TGF-β1-induced pulmonary fibroblasts,compared with control,pulmonary fibroblast proliferation and the levels of collagen Ⅰ and Ⅲ,p-JNK,8-OHdG were all significantly increased in H2O2 and TGF-β1 groups; while these changes were markedly blocked with the treatment of NAC.Conclusion TGF-β1 induces pulmonary fibroblasts to generate ROS,which contributes to JNK activation and pulmonary fibroblast proliferation as well as collagen synthesis,while ROS inhibition suppresses this effet of TGF-β1 in pulmonary fibroblasts.