Wu Kun-Fei, Fang Yang, Li Dan-Dan, Zhang Li-Juan, Li Qian, Wang Rui-Min
North China Coal Medical College, Tangshan 063000, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2009 Jul;27(7):385-9.
To investigate the role of extracellular signal-regulated kinase 1/2 on the inhibition of N-acetyl-seryl-aspartyl-lysyl-proline (AcSDKP) on the proliferation and collagen synthesis of cultured rat pulmonary fibroblasts induced by platelet-derived growth factor (PDGF).
Pulmonary fibroblasts were prepared from lungs of neonatal Wistar rats as described previously. Cells were divided into 4 groups: (1) control group (0.4% FBS group); (2) PDGF (10 ng/ml) stimulated group; (3) PD98059+PDGF group (25 micromol/L PD98059+10 ng/ml PDGF); (4) AcSDKP+PDGF group (10(-8) mol/L AcSDKP+10 ng/ml PDGF). All experiments were performed in the fourth passages. Metabolic activity of fibroblasts was observed by MTT, and expressions of type I and type III collagen were measured by immunocytochemistry and western blot. Expressions of phospho-ERK1/2 and ERK1/2 were detected by western blot.
Compared with control group, exposure of pulmonary fibroblasts to 10 ng/ml PDGF increased cell metabolic activity, expression of type I and type III collagen and phosphorylation of ERK1/2. 25 micromol/L PD98059 and AcSDKP both could inhibit the metabolic activity of pulmonary fibroblasts, type I and type III collagen synthesis and phosphorylation of ERK1/ 2 induced by PDGF, with significant differences (P < 0.05). AcSDKP+PDGF group compared with PDGF stimulated group, metabolic activity of pulmonary fibroblasts decreased to 77.4%. Immunocytochemistry result showed that in AcSDKP+PDGF group, expressions of type I and type III collagen decreased to 69.3% and 67.2% compared with those of PDGF stimulated group. Western blot result showed that in AcSDKP+PDGF group, expressions of type I and type III collagen decreased to 92.4% and 78.0%, phospho-ERK1/2 decreased to 83.5% compared with those of PDGF stimulated group, with significant differences (P < 0.05).
ERK1/2 plays an important role in the inhibition of AcSDKP on the proliferation and collagen synthesis of cultured rat pulmonary fibroblasts induced by PDGF.
探讨细胞外信号调节激酶1/2在N - 乙酰 - 丝氨酰 - 天冬氨酰 - 赖氨酰 - 脯氨酸（AcSDKP）抑制血小板衍生生长因子（PDGF）诱导的大鼠肺成纤维细胞增殖及胶原合成中的作用。
按照先前描述的方法从新生Wistar大鼠肺中制备肺成纤维细胞。细胞分为4组：（1）对照组（0.4%胎牛血清组）；（2）PDGF（10 ng/ml）刺激组；（3）PD98059 + PDGF组（25 μmol/L PD98059 + 10 ng/ml PDGF）；（4）AcSDKP + PDGF组（10⁻⁸ mol/L AcSDKP + 10 ng/ml PDGF）。所有实验均在第四代进行。通过MTT观察成纤维细胞的代谢活性，通过免疫细胞化学和蛋白质印迹法检测Ⅰ型和Ⅲ型胶原的表达。通过蛋白质印迹法检测磷酸化ERK1/2和ERK1/2的表达。
与对照组相比，肺成纤维细胞暴露于10 ng/ml PDGF可增加细胞代谢活性、Ⅰ型和Ⅲ型胶原表达以及ERK1/2磷酸化。25 μmol/L PD98059和AcSDKP均可抑制PDGF诱导的肺成纤维细胞代谢活性、Ⅰ型和Ⅲ型胶原合成以及ERK1/2磷酸化，差异有统计学意义（P < 0.05）。AcSDKP + PDGF组与PDGF刺激组相比，肺成纤维细胞代谢活性降至77.4%。免疫细胞化学结果显示，AcSDKP + PDGF组中，Ⅰ型和Ⅲ型胶原表达与PDGF刺激组相比分别降至69.3%和67.2%。蛋白质印迹结果显示，AcSDKP + PDGF组中，Ⅰ型和Ⅲ型胶原表达与PDGF刺激组相比分别降至92.4%和78.0%，磷酸化ERK1/2降至83.5%，差异有统计学意义（P < 0.05）。
ERK1/2在AcSDKP抑制PDGF诱导的大鼠肺成纤维细胞增殖及胶原合成中起重要作用。
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