Angiotensin II induces hypertrophy of human airway smooth muscle cells: expression of transcription factors and transforming growth factor-beta1
Increased smooth muscle mass due to hyperplasia and hypertrophy of airway smooth muscle (ASM) cells is a common feature in asthma. Angiotensin II (Ang II), a potent vasoconstrictor and mitogen for a wide variety of cells, has recently been implicated in bronchoconstriction in asthmatics. However, a possible mitogenic role as well as underlying molecular mechanisms of this octapeptide in human ASM cells are not yet known. We studied the effects of Ang II on ASM cell proliferation and growth and on the expression of three transcription factors, egr-1, c-fos, and c-jun, as well as a cytokine, transforming growth factor-beta1 (TGF-beta1). Human ASM cells were isolated by enzymatic digestion of bronchial smooth muscle obtained from lung resection tissue. Confluent cells were growth-arrested and subsequently incubated with Ang II (100 nM) for different time periods and processed for the measurement of cell growth and gene expression. Ang II significantly induced DNA and protein synthesis in human ASM cells at 8 h, resulting in a net increase in the accumulation of protein over DNA (i.e., cellular hypertrophy) at 16 h of incubation. Cell counts and MTT-reduction assay, however, showed no increase in cell number as a result of Ang II stimulation. Ang II stimulated the expression of egr-1 and c-fos as early as 15 min, reaching maximum levels at 45 min, whereas the expression of c-jun peaked at 2 h of Ang II exposure. Furthermore, steady-state mRNA levels of TGF-beta1 were upregulated by Ang II after 4 h and reached peak levels at 16 h of incubation. Secretion of biologically active TGF-beta1 from human ASM cells was significantly (P <= 0.02) enhanced by Ang II incubation after 8 h, which remained elevated until 24 h. Our results suggest that the Ang II-induced transient early expression of transcription factors may regulate autocrine genes like TGF-beta1, of which the subsequent late upregulation could contribute to cellular hypertrophy during, for example, airway remodeling in asthma.
|*Immediate-Early Proteins, Angiotensin II/*pharmacology, Blotting, Northern, Bronchi/drug effects/*metabolism/pathology, Cells, Cultured, DNA-Binding Proteins/metabolism, Early Growth Response Protein 1, Humans, Hyperplasia/chemically induced/metabolism, Hypertrophy/chemically induced/metabolism, Immunohistochemistry, Muscle, Smooth/drug effects/*metabolism/*pathology, Proto-Oncogene Proteins c-fos/metabolism, Proto-Oncogene Proteins c-jun/metabolism, RNA, Messenger/analysis, Research Support, Non-U.S. Gov't, Time Factors, Transcription Factors/*metabolism, Transforming Growth Factor beta/*metabolism|
|American Journal of Respiratory Cell and Molecular Biology|
|Organisation||Erasmus MC: University Medical Center Rotterdam|
McKay, S, de Jongste, J.C, Saxena, P.R, & Sharma, H.S. (1998). Angiotensin II induces hypertrophy of human airway smooth muscle cells: expression of transcription factors and transforming growth factor-beta1. American Journal of Respiratory Cell and Molecular Biology. Retrieved from http://hdl.handle.net/1765/8834