Pentoxifylline attenuates transforming growth factor-beta1-stimulated elastogenesis in human tunica albuginea-derived fibroblasts part 2: Interference in a TGF-beta1/Smad-dependent mechanism and downregulation of AAT1.
BACKGROUND
Transforming growth factor-beta1 (TGF-beta1) contributes to the pathogenesis of Peyronie's disease (PD). Pentoxifylline (PTX) antagonizes the effects of TGF-beta1 and has been utilized in our clinic for the management of PD although the mechanisms of action are not entirely clear.


OBJECTIVE
We studied cell-signaling pathways through which TGF-beta1 and PTX mediate collagen metabolism, elastin expression, and elastogenesis in tunica albuginea-derived fibroblasts (TADFs).


METHODS
TADFs from men with and without PD were cultured and treated with TGF-beta1 and PTX as monotherapy at differing concentrations and time points. Combination treatment (TGF-beta1 followed by PTX and vice versa) was also investigated.


METHODS
Reverse-transcription polymerase chain reaction and Western blotting were utilized to assess differences in elastin metabolism and cellular signaling between groups. Alpha-1 antitrypin (AAT1) expression was assayed.


RESULTS
At doses greater than 0.1 ng/Ml, TGF-beta1 increased messenger ribonucleic acid (mRNA) and protein expression of elastin in a time-dependent fashion in TADF. PTX did not interfere with TGF-beta1 mediated upregulation of elastin mRNA and protein in TADF. However, pretreatment of TADF with PTX was associated with decreased expression of AAT1, decreased activity of the Smad1/5 pathway, and enhanced phosphorylation of the inhibitory Smad6.


CONCLUSIONS
Expression of elastin mRNA and protein is upregulated in TADF by TGF-beta1. PTX has no effect on elastin production but attenuates elastogenesis in TADF through an AAT1-related mechanism.
Lin
Guiting
G
Knuppe Molecular Urology Laboratory, Department of Urology, University of California, San Francisco, CA 94143-0738, United States. glin@urology.ucsf.edu
Shindel
Alan W
AW
Banie
Lia
L
Ning
Hongxiu
H
Huang
Yun-Ching
YC
Liu
Gang
G
Lin
Ching-Shwun
CS
Lue
Tom F
TF
D015153:Blotting, Western; D002352:Carrier Proteins / Q000502:physiology*; D003094:Collagen / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects*; D004547:Elastic Tissue / Q000187:drug effects* / Q000503:physiopathology; D004549:Elastin / Q000235:genetics / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects*; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology; D006801:Humans; D008297:Male; D010411:Penile Induration / Q000503:physiopathology*; D010413:Penis / Q000187:drug effects*; D010431:Pentoxifylline / Q000494:pharmacology*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D051785:Smad Proteins / Q000502:physiology*; D053773:Transforming Growth Factor beta1 / Q000037:antagonists & inhibitors* / Q000502:physiology*
C470011:CFAP91 protein, human; D002352:Carrier Proteins; D010726:Phosphodiesterase Inhibitors; D012333:RNA, Messenger; D051785:Smad Proteins; D053773:Transforming Growth Factor beta1; D003094:Collagen; D004549:Elastin; D010431:Pentoxifylline
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