Zhao Lishuang, Wei Zhongqiu, Yang Fang, Sun Ying
Pathology Department, College of Basic Medical Science, Hebei United University, Tanshan 063000, China.
Pathology Department, College of Basic Medical Science, Hebei United University, Tanshan 063000, China. E-mail:
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2015 Jan;33(1):15-9.
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.
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.
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 II, 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.
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.
本研究将探讨活性氧（ROS）是否参与转化生长因子-β1（TGF-β1）诱导的JNK激活、肺成纤维细胞增殖以及I型和III型胶原蛋白合成。
将肺成纤维细胞随机分为对照组（0.4%血清）和TGF-β1（5μg/L）组，以检测TGF-β1是否能诱导肺成纤维细胞增殖、I型和III型胶原蛋白合成、磷酸化JNK（p-JNK）和8-羟基脱氧鸟苷（ROS指标）；而在探索N-乙酰半胱氨酸（NAC，抗氧化剂）对TGF-β1诱导的肺成纤维细胞是否具有抑制作用的部分实验中，设置了对照组（0.4%血清）、过氧化氢（H2O2，0.1mmol/L，阳性对照）、H2O2+NAC（10mmol/L）、TGF-β1（5μg/L）、TGF-β1+NAC组。分别采用MTT法、免疫荧光法和蛋白质印迹法检测肺成纤维细胞增殖、8-羟基脱氧鸟苷水平、JNK以及I型和III型胶原蛋白的表达。
在检测TGF-β1对肺成纤维细胞作用的实验中，与对照组相比，TGF-β1显著刺激肺成纤维细胞增殖，并增加I型和III型胶原蛋白、p-JNK和8-羟基脱氧鸟苷水平。在接下来探索NAC对TGF-β1诱导的肺成纤维细胞是否具有抑制作用的实验中，与对照组相比，H2O2组和TGF-β1组的肺成纤维细胞增殖以及I型和II型胶原蛋白、p-JNK、8-羟基脱氧鸟苷水平均显著升高；而NAC处理可明显阻断这些变化。
TGF-β1诱导肺成纤维细胞产生活性氧，这有助于JNK激活、肺成纤维细胞增殖以及胶原蛋白合成，而抑制活性氧可抑制TGF-β1在肺成纤维细胞中的这种作用。
