title: Repair of protease-damaged elastin in neonatal rat aortic smooth muscle cell cultures.
abstract: The objective of this study was to investigate the elastin repair process in the rat aortic smooth muscle cell culture after proteolytic injury. Although little studied in vivo, elastin repair is thought to occur through a sequential process involving enzymatic removal (debridement) of damaged fibers followed by synthesis of tropoelastin, its subsequent processing, and eventual incorporation into new insoluble elastin. A second repair mechanism of proteolytically damaged elastin in a culture system is reported here. Repair in this system relates directly to restoration of resistance to elastin solubilization by hot alkali. As expected, severe injuries were observed with porcine pancreatic elastase (PPE). Using PPE, only 6% of the elastin, relative to control, was resistant to hot alkali immediately after elastase treatment. 4 wk later, resistance to hot alkali had increased dramatically to a mean of 90%. Repair took longer after injury with 75 micrograms of PPE as compared with 50 micrograms of PPE. The limited elastic fiber proteolysis induced by either human neutrophil elastase or porcine trypsin was repaired in culture within 2 wk. Elastin that had been radiolabeled with [3H]lysine 4-5 wk before injury was converted from a hot NaOH-susceptible to a NaOH-resistant elastin fraction during recovery from PPE injury. At the same time, the frayed elastic fibers that were seen with the electron microscope immediately after PPE treatment were replaced by continuous bands of elastin that resembled those in control cultures. Restoration of NaOH resistance did not require a net increase in total cell layer elastin, suggesting that relatively little new tropoelastin incorporation into the cell layer was required for this type of repair. These results suggested a salvage repair mechanism for proteolytically damaged elastin.
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D004549:Elastin / Q000378:metabolism* / Q000648:ultrastructure; D006863:Hydrogen-Ion Concentration; D008239:Lysine / Q000493:pharmacokinetics; D008854:Microscopy, Electron; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012972:Sodium Hydroxide / Q000494:pharmacology; D012995:Solubility
chemical_list: D012972:Sodium Hydroxide; D004549:Elastin; D010447:Peptide Hydrolases; D008239:Lysine