sirolimus and pirfenidone

Topics: Anti-Bacterial Agents; Anti-Inflammatory Agents, Non-Steroidal; Doxycycline; Enzyme Inhibitors; Humans; Idiopathic Pulmonary Fibrosis; Immunosuppressive Agents; Indoles; Lung Diseases, Interstitial; Lymphangioleiomyomatosis; Pyridones; Sarcoidosis, Pulmonary; Sirolimus; Smoking; Tomography, X-Ray Computed; Tumor Necrosis Factor-alpha

Pirfenidone, a pleiotropic anti-fibrotic treatment, has been shown to slow down disease progression of idiopathic pulmonary fibrosis (IPF), a fatal and devastating lung disease. Rapamycin, an inhibitor of fibroblast proliferation could be a potential anti-fibrotic drug to improve the effects of pirfenidone.. Primary lung fibroblasts from IPF patients and human alveolar epithelial cells (A549) were treated in vitro with pirfenidone and rapamycin in the presence or absence of transforming growth factor β1 (TGF-β). Extracellular matrix protein and gene expression of markers involved in lung fibrosis (tenascin-c, fibronectin, collagen I [COL1A1], collagen III [COL3A1] and α-smooth muscle actin [α-SMA]) were analyzed. A cell migration assay in pirfenidone, rapamycin and TGF-β-containing media was performed.. Gene and protein expression of tenascin-c and fibronectin of fibrotic fibroblasts were reduced by pirfenidone or rapamycin treatment. Pirfenidone-rapamycin treatment did not revert the epithelial to mesenchymal transition pathway activated by TGF-β. However, the drug combination significantly abrogated fibroblast to myofibroblast transition. The inhibitory effect of pirfenidone on fibroblast migration in the scratch-wound assay was potentiated by rapamycin combination.. These findings indicate that the combination of pirfenidone and rapamycin widen the inhibition range of fibrogenic markers and prevents fibroblast migration. These results would open a new line of research for an anti-fibrotic combination therapeutic approach.

Topics: A549 Cells; Alveolar Epithelial Cells; Biomarkers; Cell Movement; Epithelial-Mesenchymal Transition; Extracellular Matrix; Humans; Idiopathic Pulmonary Fibrosis; Myofibroblasts; Pyridones; Sirolimus; Transforming Growth Factor beta1

Benign biliary stricture (BBS) is highly refractory. Currently, there is no effective strategy for prevention of BBS recurrence. The aim of this study is to establish a novel BBS rabbit model and to investigate the efficacy of biliary infusion with anti-proliferative medications for treating BBS.. A BBS model was established via surgical injury and biliary infection. The biliary infusion tube was inserted into the common bile duct via the stump of cystic duct after cholecystectomy. Biliary infusions with Rapamycin, Pirfenidone and Fasudil were performed daily during the 4 weeks following the surgery. The wall thickness and luminal area of the bile duct were assessed.. All rabbits formed BBS after surgery. The mortality rate was 13% (8/60) and tube withdrawal rate was 4% (2/48). The thickness of the bile duct wall was significantly reduced; whereas the luminal area of the bile duct was dramatically enlarged in the Rapamycin or the Pirfenidone treated group, compared to the saline treated group. Furthermore, the local treatment significantly decreased the levels of proliferation makers, including PCNA, Collagen I and fibrogenic mediators, including ACTA2 and TGF-beta.. We have established a novel animal model for BBS formation. We have further demonstrated that biliary infusion with Rapamycin or Pirfenidone limits the biliary strictures through inhibiting the proliferation of the bile duct wall in this model. This may represent a new avenue for preventing biliary restenosis.

Topics: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; Animals; Antineoplastic Agents; Biliary Tract Diseases; Constriction, Pathologic; Disease Models, Animal; Drug Evaluation, Preclinical; Pyridones; Rabbits; Secondary Prevention; Sirolimus; Vasodilator Agents

Currently one of the major problems that interventional pulmonologists have to face is the increased proliferation of fibrinous tissue on the site of the stent placement, and usually at the two ends.. The drugs rapamycin and pirfenidone were chosen for our experiment. Fibroblasts were also cultured in order to administer pirfenidone and rapamycin in different concentrations. The following cell viability methods were used: (a) Senescence - Cell Titer Assay, (b) Necrosis - Cyto Tox Assay and (c) Apoptosis - Caspase-Glo 3/7 Assay.. Rapamycin has minimal to no effect on fibroblasts regarding apoptosis, senescence and necrosis. 0.1 to 1 μM. Pirfenidone concentrations lead to an elevated cell metabolism because cells try to evade the cytotoxic effect of the drug. Increasing Pirfenidone concentrations lead to higher apoptosis rates. 10 μM pirfenidone induces the highest apoptosis rates in this experiment and reduce cell viability to a minimum.. Necrosis is unaffected by the investigated drugs.

Topics: Apoptosis; Caspase 3; Caspase 7; Cell Line; Cell Survival; Drug-Eluting Stents; Fibroblasts; Humans; Immunosuppressive Agents; L-Lactate Dehydrogenase; Pyridones; Sirolimus

We previously showed that pirfenidone, an anti-fibrotic agent, reduces lung allograft injury or rejection. In this study, we tested the hypothesis that pirfenidone has immune modulating activities and evaluated its effects on the function of T-cell subsets, which play important roles in allograft rejection.. We first evaluated whether pirfenidone alters T-cell proliferation and cytokine release in response to T-cell receptor (TCR) activation, and whether pirfenidone alters regulatory T cells (CD4CD25) suppressive effects using an in vitro assay. Additionally, pirfenidone effects on alloantigen-induced T-cell proliferation in vivo were assessed by adoptive transfer of carboxyfluorescein diacetate succinimidyl ester-labeled T cells across a parent->F1 major histocompatibility complex mismatch, as well as using a murine heterotopic cardiac allograft model (BALB/c->C57BL/6).. Pirfenidone was found to inhibit the responder frequency of TCR-stimulated CD4 cell total proliferation in vitro and in vivo, whereas both CD4 and CD8 proliferation index were reduced by pirfenidone. Additionally, pirfenidone inhibited TCR-induced production of multiple pro-inflammatory cytokines and chemokines. Interestingly, there was no change on transforming growth factor-beta production by purified T cells, and pirfenidone had no effect on the suppressive properties of naturally occurring regulatory T cells. Pirfenidone alone showed a small but significant (P<0.05) effect on the in vivo allogeneic response, whereas the combination of pirfenidone and low dose rapamycin had more remarkable effect in reducing the alloantigen response with prolonged graft survival.. Pirfenidone may be an important new agent in transplantation, with particular relevance to combating chronic rejection by inhibiting both fibroproliferative and alloimmune responses.

Topics: Adoptive Transfer; Animals; CD8-Positive T-Lymphocytes; Cell Proliferation; Cells, Cultured; Chemokines; Cytokines; Drug Therapy, Combination; Graft Rejection; Heart Transplantation; Histocompatibility; Immunosuppressive Agents; Isoantigens; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Pyridones; Sirolimus; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory; Time Factors

Obliterative bronchiolitis (OB) is the histologic correlate of chronic airway rejection, which remains the most significant cause of death in long-term survivors of lung transplantation. Using an established murine heterotopic tracheal transplant model of chronic airway rejection, the effects of the oral anti-fibrotic agent pirfenidone on development of the OB-like lesion were evaluated.. Tracheas from BALB/c mice were implanted into the sub-cutaneous tissue of C57BL/6 mice, and the allografts were evaluated morphologically for airway rejection changes and immunohistochemically for transforming growth factor (TGF)-beta at 16 or 28 days after transplantation. In addition, the potential additive effects of pirfenidone in combination with 2 immunosuppressive agents, cyclosporine or rapamycin, was evaluated.. Compared with untreated controls, pirfenidone-fed mice showed less epithelial cell injury and luminal granulation tissue and fibrosis. Plasma TGF-beta levels and local TGF-beta expression based on immunohistochemistry were decreased in the pirfenidone-treated animals. Pirfenidone given on Day 9 or 16 post-transplant through Day 28 resulted in no significant improvement compared with controls. There was no significant additive effect of pirfenidone in combination with cyclosporine, whereas pirfenidone plus rapamycin demonstrated additive protection against the development of the obstructive airway lesion.. In aggregate, these results show that the anti-fibrotic agent pirfenidone inhibits the development of the OB-like lesion in this mouse model of human chronic airway rejection, and that these effects may be mediated by TGF-beta. The results also suggest that pirfenidone may be worthy of investigation in human lung transplant recipients at high risk of developing OB.

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents, Non-Steroidal; Chronic Disease; Cyclosporine; Disease Models, Animal; Female; Graft Rejection; Immunosuppressive Agents; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Pyridones; Sirolimus; Trachea; Transforming Growth Factor beta