Fenfluramine-Phentermine is Associated with an Increase in Cellular Proliferation Ex Vivo and In Vitro

Abstract

Background and aim of the study: Fenfluraminephentermine (FenPhen) has been implicated in accelerated valvular heart disease, characterized by valvular regurgitation and thickening, and resembling the histopathologic lesions found in carcinoid. The study aim was to determine whether cellular proliferation is present in FenPhen-exposed valves, by utilizing an in-vitro model to test whether FenPhen has a direct mitogenic effect on cardiac valvular cells, as compared to serotonin.

Methods: Ex-vivo valves were tested for proliferation in surgically removed FenPhen-exposed valves (n = 10) and compared to proliferation levels in normal human cardiac valves removed at autopsy (n = 10). Immunostaining for a DNA polymerase, proliferating cell nuclear antigen (PCNA), was performed and quantified using digital imaging analysis. In-vitro assays were performed for direct proliferative effects of serotonin and FenPhen (10-6, 10-7 and 10-8 M) on porcine aortic valve subendothelial cells, using a [3H]-thymidine incorporation assay.

Results: Ex-vivo PCNA levels in human FenPhenexposed valves were elevated compared to controls (22.8 ± 4.54 versus 1.26 ± 0.47; p <0.001). In vivo, serotonin and FenPhen markedly increased (10-fold) cell proliferation (as measured by [3H]-thymidine incorporation) in subendothelial cells in vitro (p <0.001). This proliferative response was demonstrated by PCNA staining in carcinoid heart valves and FenPhen-exposed valves. Mechanistically, plateletderived growth factor increased cell proliferation in a dose-related manner (p <0.001), the response being inhibited by a MAP kinase inhibitor (determined by monitoring p42/44 levels).

Conclusions: In vitro, FenPhen acts as a powerful mitogen on subendothelial myofibroblast valve cells. Ex vivo, cellular proliferation was significantly elevated in human FenPhen-exposed cells.