摘要
目的:本研究观察血管紧张素II(angiotensin II,AngII)对增生性瘢痕成纤维细胞细胞外信号调节激酶(extracellularsignal-regulated kinase,ERK)活性的影响,旨在探讨其影响增生性瘢痕成纤维细胞生物学行为的信号机制。方法:取自愿捐献的增生性瘢痕组织标本,采用胶原酶法行成纤维细胞培养。用免疫荧光组织化学法检测细胞AT1和AT2受体的表达。取第3～4代细胞,并按实验设计,分别加入10-7mol/LAngII,10-7mol/LAngII+10μmol/LValsartan,10-7mol/LAngII+10μmol/LPD123319,10μmol/LValsartan,10μmol/LPD123319共5组;对照组仅加入等量DMEM。以Western Blot法检测培养的细胞细胞外信号调节激酶(extracellular signal-regulated kinases,ERK)活性变化,观察在阻断AT1或AT2受体情况下AngII对培养的瘢痕成纤维细胞ERK磷酸化的影响。结果:免疫荧光组织化学染色结果显示培养的增生性瘢痕成纤维细胞同表达AT1和AT2受体。AngII可增加培养的增生性瘢痕成纤维细胞ERK磷酸化。在一定剂量范围内,AngII浓度增加,细胞ERK磷酸化程度增加。与对照组比较10-7mol/LAngII显著增加瘢痕成纤维细胞ERK磷酸化,且差异有统计学意义(P<0.05)。10μmol/LAT2受体拮抗剂PD1233191可显著增强AngII诱导的细胞ERK磷酸化(P<0.05);10μmol/L的AT1受体拮抗剂Valsartan可显著抑制AngII诱导的细胞ERK磷酸化;Valsartan或PD1233191单独刺激细胞并未明显影响ERK磷酸化(P>0.05)。应用抗ERK抗体显示各组ERK含量一致。结论:AngII通过其受体AT1和AT2可调控增生性瘢痕成纤维细胞ERK磷酸化,AngII对增生性瘢痕成纤维细胞ERK活性的影响可能是AngII调控增生性瘢痕成纤维细胞生物学行为可能的信号机制之一。
Objective The present study was conducted to observe the effect of angiotensin Ⅱ （angiotensin Ⅱ, Ang Ⅱ） on the phosphorylation of extracellular signal-regulated kinase （ERK） of fibroblasts from hyertrophic scar, and explore the possible signaling mechanism whereby Ang Ⅱ influences biological behaviors of hypertrophic scar fibroblasts. Methods The expression of AT1 and AT2 receptor was detected by immunofluorescence staining. Cultured human skin fibroblasts were treated with Ang Ⅱ （10^-7mol/L）, with or without an AT1 receptor blocker, valsartan or an AT2 receptor antagonist, PD123319. The phosphorylation of ERK was detected by western blot. Results Immunofluorescence staining showed that cultured fibroblasts derived from hypretrophic scars coexpressed both AT1 and AT2 receptors. Ang Ⅱ increased ERK phosphorylation in cultured hypertrophic scar fibroblasts dose- and time-dependently. In addition, the Ang Ⅱ-induced ERK phosphorylation was markedly inhibited by valsartan, an AT1 receptor specific blocker （P〉0.05）, whereas enhanced by PD123319, an AT2 receptor antagonist （P〈0.05）. Valsartan or PD123319 did not influence ERK phosphorylation of fibroblasts derived from hypertrophic scars （P〉0.05）. Total protein levels of ERK did not differ in each experimental group. Conclusion These results indicate that Ang Ⅱ via AT1 and AT2 receptors regulates ERK phosphorylation of hypertrophic scars fibroblasts, thereby influencing biological behavior of fibroblasts derived from hypertrophic scar. These findings may help in understanding the molecular mechanism of human hypertrophic scar formation.
出处
《中国美容医学》
CAS
2007年第7期874-877,共4页
Chinese Journal of Aesthetic Medicine
基金
国家重点基础研究发展规划资助项目(2005CB522603)
