PTX3 Controls Activation of Matrix Metalloproteinase 1 and Apoptosis in Conjunctivochalasis Fibroblasts

Abstract

PURPOSE. Conjunctivochalasis (CCh) is an age-related inflammatory ocular surface disease manifesting redundant, loose conjunctiva folds. The pathogenic role of Pentraxin 3 (PTX3) in controlling upregulation of matrix metalloproteinase 1 (MMP-1) and MMP-3 in CCh remains undefined.METHODS. Cytolocation of PTX3 and apoptosis were compared by immunostaining and terminal deoxyribonucleotidyl transferase-mediated FITC-linked dUTP nick-end DNA labeling (TUNEL) assay between normal and CCh specimens containing the conjunctiva and the Tenon. Second to third cultures of normal and CCh fibroblasts were treated with or without Aprotinin, Batimastat, or N-isobutyl-N-(4-methoxyphenylsulfonyl)-glycylhydroxamic acid (NNGH), followed by transfection with or without PTX3 siRNA, and TNF-a or IL-1b. Cell lysates and culture media were collected to assess apoptosis measured by the Cell Death Detection ELISA and expression of PTX3, MMP-1, and MMP-3 transcripts and proteins by quantitative RT-PCR and Western blot, respectively. RESULTS. PTX3 immunostaining was negative in normal specimens, but strongly positive in the subconjunctival stroma of CCh specimens. More apoptotic cells were found in CCh samples than in normal specimens. Expression of PTX3 transcripts and protein was not constitutive in resting normal fibroblasts but was in resting CCh fibroblasts and was upregulated by IL-1b in both cell lysates and culture media of both fibroblasts. PTX3 siRNA further upregulated MMP-1 and MMP-3 transcripts in resting normal fibroblasts, but synergistically with IL-1b upregulated the expression of MMP-1 and MMP-3 transcripts only in CCh fibroblasts, with activation of MMP-1 more so than MMP-3. PTX3 siRNA knockdown also promoted cell death characterized by apoptosis and necrosis, and such cell death could be rescued by inhibitors against serine proteinase, MMP1, or MMP3.CONCLUSIONS. Perturbation of PTX3 expression might partake in apoptosis and pathogenesis of CCh by upregulating expression of MMP-1 and MMP-3, and activation of MMP-1 and MMP-3.

Potent Vasoconstrictor Kisspeptin‐10 Induces Atherosclerotic Plaque Progression and Instability: Reversal by its Receptor GPR54 Antagonist

“…Pentraxin‐3 is well known to reflect vascular inflammation derived from mainly ECs followed by VSMCs and macrophages and also is associated with apoptosis and plaque vulnerability . In the present study, increased pentraxin‐3 expression and increased activities of MMP‐2 and MMP‐9 as well as apoptosis induction in VSMCs by KP‐10 may be associated with atherosclerotic plaque instability.…”

“…Pentraxin-3 is well known to reflect vascular inflammation derived from mainly ECs followed by VSMCs and macrophages and also is associated with apoptosis and plaque vulnerability. [33][34][35][36] In the present study, increased pentraxin-3 expression and increased activities of MMP-2 and MMP-9 as well as apoptosis induction in VSMCs by KP-10 may be associated with atherosclerotic plaque instability. Previous reports showed that KPs downregulated MMP-2 and MMP-9 at both the transcriptional and protein levels in human renal carcinoma KU19-20 cells and fibrosarcoma HT-1080 cells, respectively.…”

Clinical characteristics of patients with conjunctivochalasis

“…They stated that pentraxin 3 expression might partake in apoptosis and pathogenesis of CCh by upregulating expression of matrix metalloproteinase-1 (MMP-1) and matrix metalloproteinase-3 (MMP-3). In another study of Guo et al19 the authors demonstrated that dysfunction of tumor necrosis factor-stimulated gene-6 might play a role in the pathogenesis of CCh by counteracting the transcription of MMP-1 and MMP-3 and the activation of MMP-1.…”

Tunicamycin-induced Endoplasmic Reticulum Stress Upregulates the Expression of Pentraxin 3 in Human Retinal Pigment Epithelial Cells

“…Several studies have demonstrated that PTX3 is involved in the removal of apoptotic cells during immune response [101112]. PTX3 siRNA knockdown was also shown to promoted cell death, characterized as apoptosis and necrosis, in the presence of proinflammatory cytokines such as IL-1β and TNF-α in conjunctivochalasis fibroblasts [13]. …”