Effect of biological amniotic membrane soaked in pirfenidone on scar formation after glaucoma filtration surgery in rabbit eyes / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the application of biological amniotic membrane soaked in pirfenidone(PFD)and to evaluate its anti-scarring effect and toxic side effects on glaucoma model of rabbit eyes.METHODS: The right eyes of 72 healthy New Zealand white rabbits were randomly divided into 0.5%PFD+ biological amniotic membrane group, biological amniotic membrane group, mitomycin C(MMC)group and blank control group after the glaucoma model was established by anterior chamber injection of compound carbomer solution, and 18 rabbits in each group underwent trabeculectomy, in which the 0.5% PFD+ biological amniotic membrane group was placed with 0.5% PFD solution-soaked biological amniotic membrane under the scleral flap, and the biological amniotic membrane group was placed with normal saline-soaked rehydrated biological amniotic membrane under the scleral flap. In the MMC group, a cotton pad soaked in MMC was placed under the scleral flap for 3 min and immediately rinsed with normal saline, while the blank control group received no implant after the scleral flap was made. The intraocular pressure(IOP), filtration blebs, toxic side effects and complications were evaluated, and the histopathological changes in the filtration area were observed by hematoxylin-eosin(HE), Masson staining and immunohistochemical staining.RESULTS: The mean IOP at 14, 21 and 28 d after trabeculectomy were 0.5%PFD+ biological amniotic membrane group<MMC group<biological amniotic membrane group<blank control group(all P<0.05). At 28 d after trabeculectomy, 0.5%PFD+ biological amniotic membrane group had the best effect of anti-inflammatory hyperplasia and inhibition of collagen formation, the highest survival rate of filtration blebs, and the inflammatory reaction was mild.CONCLUSION: Biological amniotic membrane soaked in pirfenidone has more obvious anti-scarring effect on glaucoma model, with less toxic side effects and good safety.

Effect of pirfenidone on paraquat-induced pulmonary fibrosis in rats / 中华劳动卫生职业病杂志

Objective: To construct paraquat (PQ) poisoning rat model and to explore the effect of pirfenidone (PFD) on PQ-induced pulmonary fibrosis. Methods: In April 2017, male 6-8 week-old Wistar rats were selected, and PQ was administered intraperitoneally at one time. PFD was administered by gavage 2 hours after poisoning. The daily gavage doses were 100, 200 and 300 mg/kg, and the rats were divided into physiological saline group, PQ group, PQ+PFD 100 group, PQ+PFD 200 group, PQ+PFD 300 group, with 10 rats in each group at each observation time point. The pathological changes of lung tissue at different time points (the 1st, 3rd, 7th, 14th, 28th, 42nd and 56th days) after poisoning and the effect of PFD intervention with different dose on PQ-induced pulmonary fibrosis were observed. Pathological evaluation of lung tissue was performed by Ashcroft scale method. The PQ+PFD 200 group was selected to further explore the pathological changes of lung tissue, the contents of hydroxyproline and malondialdehyde in lung tissue were determined.And the tumor necrosis factor (TNF) -α, interleukin (IL) -6, transforming growth factor (TGF) -β1, fibroblast growth factor (FGF) -B, platelet-derived growth factor (PDGF) -AB, insulin-like growth factor (IGF) -1 and PQ concentrations in serum and lung tissue were determined. Results: On the 1st to 7th day after PQ exposure, rats developed lung inflammation, which was aggravated on the 7th to 14th day, and pulmonary fibrosis appeared on the 14th to 56th day. Compared with PQ group, the Ashcroft scores of lung fibrosis in PQ+PFD 200 group and PQ+PDF 300 group decreased significantly in 7th and 28th day (P<0.05), while the Ashcroft score of lung fibrosis in PQ+PFD 100 group had no significant difference (P>0.05). After PQ exposure, the content of hydroxyproline in lung tissue increased gradually and reached the peak value on the 28th day. Compared with the PQ group, the contents of hydroxyproline in the PQ+PFD 200 group decreased at the 7th, 14th and 28th day, and the contents of malondialdehyde decreased at the 3rd and 7th day, the differences were statistically significant (P<0.05). The levels of TNF-α, IL-6 in rat serum and lung tissue reached the peak value on the 7th day after PQ exposure, and the levels of TGF-β1, FGF-B and IGF-1 in rat serum and lung tissue reached the peak value on the 14th day after PQ exposure, and the level of PDGF-AB in rat serum and lung tissue reached the peak value on the 28th day after PQ exposure. Compared with PQ group, the level of serum IL-6 in PQ+PFD 200 group decreased significantly on the 7th day, and serum TGF-β1, FGF-B, PDGF-AB and IGF-1 on the 14th and 28th day were decreased significantly (P<0.05). The levels of TNF-α, IL-6 in lung tissue of rats in PQ+PFD 200 group on the 7th day decreased significantly, and the levels of TGF-β1, FGF-B and IGF-1 in lung tissue of rats on the 14th day were significantly decreased, and the level of PDGF-AB in lung tissue of rats on the 28th day were significantly decreased (P<0.05) . Conclusion: PFD partially alleviates the PQ-induced lung inflammation and fibrosis by inhibiting oxidative stress, reducing the levels of pro-inflammatory and pro-fibrotic cytokines in serum and lung tissue, but does not affect the concentrations of PQ in serum and lung tissue.

Pirfenidone inhibits the polarization of M2 macrophages by down-regulating IRF4 to reduce the occurrence and development of radiation-induced lung fibrosis / 中华放射肿瘤学杂志

Objective:To explore the preventive and therapeutic effect of pirfenidone (PFD) on radiation-induced lung fibrosis (RILF) and its mechanism.Methods:40 female C57/BL6 mice were randomly divided into 4 groups: negative control group (NC), PFD treatment group (PFD), radiation treatment group (RT) and radiation plus PFD treatment group (RT+ PFD). Mice in RT and RT+ PFD groups received a single whole lung X-ray consisting of a 50 Gy dose of radiation, delivered by small animal radiation research platform (SARRP). PFD at a dose of 300 mg/kg was administered orally 2 h before irradiation for 150 d. HE and Masson staining were used to detect the infiltration of inflammatory cells and the degree of pulmonary fibrosis. Quantitative real-time PCR (qPCR) and Western blotting (WB) were adopted to detect the expression levels of M1/M2 macrophage phenotypic markers. The expression levels of arginase-1(ARG-1), chitinase 3-like protein 3(YM-1) and interferon regulatory factor-4(IRF4) of macrophages stimulated with IL-4 and IL-13 were detected by WB. In addition, immunofluorescence staining was used to detect the expression and translocation of IRF4 in macrophages among different treatment groups.Results:HE and Masson staining showed that PFD could significantly inhibit radiation-induced infiltration of inflammatory cells and fibrosis in lung tissues. The M2 macrophages and expression levels of ARG-1 and YM-1 were down-regulated in the RT+ PFD group. Cell experiments further confirmed that PFD could significantly inhibit the polarization of macrophages to M2 induced by IL-4+ IL-13, which was mainly related to the down-regulation of IRF4.Conclusion:PFD has a preventive and therapeutic effect on RILF by inhibiting IRF4 and reducing the polarization of macrophages to M2.

Pirfenidone alleviates urethral stricture following urethral injury in rats by suppressing TGF-β1 signaling and inflammatory response / 南方医科大学学报

OBJECTIVE@#To investigate the effect of pirfenidone for reducing urethral stricture following urethral injury in rats and explore the possible mechanism.@*METHODS@#Thirty male SD rats were randomly assigned into negative control group, positive control group and pirfenidone group (n=10). In pirfenidone and positive control groups, the rats were subjected to incision of the posterior urethral cavernous body followed by daily intraperitoneal injection of pirfenidone (100 mg/kg) and an equivalent volume of solvent, respectively. The rats in the negative control group were given intraperitoneal injections of solvent without urethral injury. At two weeks after modeling, retrograde urethrography was performed for observing urethral stricture, and the injured urethral tissues were harvested for HE staining, Masson staining, immunohistochemical staining and Western blotting for detecting the protein expressions of α-SMA and TGF-β1. The mRNA expressions of the inflammatory factors TNF-α, IL-6, and IL-1β were detected using qRT-PCR.@*RESULTS@#The body weight of the rats in pirfenidone group was significantly decreased compared with that in the other two groups (P < 0.05). Retrograde urethrography showed significant narrowing of the urethra in the positive control group but not in the pirfenidone group. HE staining of the injured urethral tissues showed obvious proliferation of urethral epithelial cells with narrow urethral cavity and increased inflammatory cells in positive control group. The pathological findings of the urethra were similar between pirfenidone group and the negative control group. Masson staining revealed obviously reduced collagen fibers and regular arrangement of the fibers in pirfenidone group as compared to the positive control group. Compared with those in the negative control group, the expressions of α-SMA and TGF-β1 were significantly increased in the positive control group, and pirfenidone treatment significantly inhibited their expressions (P < 0.05 or 0.01). Pirfenidone also significantly inhibited the mRNA expressions of TNF-α, IL-6, and IL-1β in the injured urethral tissue (P < 0.05 or 0.01).@*CONCLUSION@#Pirfenidone can prevent urethral fibrosis and stricture after urethral injury possibly by inhibiting the TGF-β1 pathway and inflammatory response.

Effect of pirfenidone on the proliferation of human glomerular mesangial cells induced by serum IgA1 of IgA nephropathy patients / 中华肾脏病杂志

Objective:To investigate the effect of pirfenidone (PFD) on the proliferation of human glomerular mesangial cells (HMC) stimulated by serum IgA1 in patients with IgA nephropathy (IgAN) and its possible mechanism.Methods:Serum IgA1 of IgAN patients was purified by Jacalin affinity chromatography combined with Sephacryl S-200 gel filtration, and then heated to aggregated form (aIgA1). CCK8 method was used to confirm the concentration and time of PFD. The cells were divided into blank control group, IgA1 (0.5 mg/ml) group and IgA1 (0.5 mg/ml)+PFD (2 mmol/L) group. The CCK8 method was used to detect proliferation of mesangial cells. The cell cycle was detected by flow cytometry, and the proliferation index of mesangial cells was calculated. The expression levels of transforming growth factor β1 (TGF-β1), Smad4, Smad7, fibronectin (FN) and collagen Ⅳ protein and mRNA were detected through Western blotting and real-time PCR.Results:Compared with blank control group, the proliferation of HMC was promoted significantly by aIgA1 ( P<0.05). After PFD treatment, the proliferation of HMC was significantly inhibited ( P<0.01). Compared with the blank control group, the number of G1 phase cells decreased, the number of S phase cells and cell proliferation index increased in IgA1 group (all P<0.05). Compared with IgA1 group, the number of cells in G1 phase increased significantly, the number of cells in S phase and G2/M phase decreased significantly, and the cell proliferation index decreased in IgA1+PFD group (all P<0.05). Western blotting and real-time PCR results showed that compared with the blank control group, the protein and mRNA expressions of collagen Ⅳ, FN and Smad4 in HMC stimulated by aIgA1 were significantly increased, while TGF-β1 protein expression was increased and Smad7 protein expression was decreased (all P<0.05). After PFD treatment, the protein and mRNA expression of collagen Ⅳ, FN and Smad4 in HMC was significantly decreased, while TGF-β1 protein expression was obviously decreased, and Smad7 protein was up-regulated (all P<0.05). There was no significant difference in the mRNA expression of TGF-β1 and Smad7 in each group before and after PFD treatment (all P>0.05). Conclusions:PFD can increase the arrest of HMC in G1 phase, inhibit the proliferation of HMC induced by aIgA1 of IgAN patients, and reduce the production of extracellular matrix. The mechanism may be related to up-regulation of Smad7 expression and down-regulation of TGF-β1/Smad4 pathway.

Study on the effect and mechanism of Pirfenidone on endothelial-mesenchymal transition in endothelial cells / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To establish the hypoxia induced endothelial-mesenchymal transition(EndoMT)model of endothelial cells, and to investigate the effect and mechanism of Pirfenidone(PFD)on inhibiting the subretinal fibrosis progression.<p>METHODS: Primary cultured human umbilical vein endothelial cells(HUVEC), 4-7 passages were used for experiments after cell identification. CoCl₂ induced hypoxia to establish the transformation model of endothelial cells into fibroblasts. CCK-8 was performed to detect cell proliferation rate and chose the optimal drug concentration. All cells were divided into 4 groups: control group(FBS-free), CoCl₂(200μmol/L)group, CoCl₂+0.3mg/mL PFD group, CoCl₂+0.6mg/mL PFD group. The protein expression of CD31, VE-cadherin, α-SMA, FSP1, p-p38 and p38 were detected by Western blot. Double immunofluorescence labeling method was used to observe the CD31/α-SMA expression. Wound healing assay detected the cell migration. The q-PCR was applied to detect the mRNA levels of TGF-β1 and SNAI1.<p>RESULTS: Compared with CoCl₂ group, PFD increased cell proliferation rate and inhibited cell migration significantly under hypoxia(<i>P</i><0.05). PFD decreased the protein expression of the mesenchymal markers α-SMA and FSP1, and increased the protein level of the endothelial markers CD31 and VE-cadherin(<i>P</i><0.05). Double immunofluorescence results showed that PFD could reduce the expression of α-SMA and increase the level of CD31(<i>P</i><0.05). In the process of EndoMT, the p38 protein expression level was stable(<i>P</i>>0.05). PFD down-regulated significantly the high protein expression of p-p38, and high mRNA expression of TGF-β1 and SNAI1 compared with control group(<i>P</i><0.05). There was no significant difference between the 0.3 and 0.6mg/mL PFD groups in all results above.<p>CONCLUSION: PFD can inhibit the formation of fibrosis in endothelial cells. TGF-β/p38MAPK signaling pathway might be one of the mechanisms that PFD regulates EndoMT progression. PFD will be expected to become a potential new sight on the treatment of subretinal fibrosis.

A Case Report of Checkpoint Inhibitor Pneumonitis Caused by PD-1 Antibody-Safety and Effectiveness of Pirfenidone / 中国肺癌杂志

BACKGROUND@#Immune checkpoint inhibitor associated pneumonia (CIP) is a serious side effect of immune checkpoint inhibitors. There is a consensus on the treatment of acute phase of CIP, but the treatment of pulmonary interstitial fibrosis after the acute phase is still a clinical problem to be solved.@*METHODS@#The diagnosis and treatment of a non-small cell lung cancer (NSCLC) patient with immune checkpoint inhibitor associated pneumonia in the Stereotactic Radiotherapy Department of Qingdao Central Hospital were retrospectively analyzed, and literatures were reviewed.@*RESULTS@#A 70-year-old male patient was diagnosed with Poorly differentiated squamous cell carcinoma of left lung with mediastinal lymph node metastasis T3N3M0 stage IIIc, EGFR/ALK/ROS1/RAF negative, PD-L1 (22c3) immunohistochemistry negative. After the progression of first-line chemotherapy, the patient was diagnosed as immune checkpoint inhibitor associated pneumonia grade 3 during second-line monotherapy with Nivolumab. After initial high-dose glucocorticoid pulse therapy, the lung computed tomography (CT) imaging and clinical symptoms of the patients were partially relieved, and then pirfenidone (300 mg tid) was given orally for more than 11 months. During the treatment of pirfenidone, the CT imaging and clinical symptoms of the patients were significantly improved, and there were no other adverse reactions except grade 1 nausea. During this period, chemotherapy and Anlotinib was given concurrently with pirfenidone and showed good safety profile.@*CONCLUSIONS@#This case report is the first report of pirfenidone in the treatment of CIP, which provides a new idea for the clinical practice and research of CIP treatment.

Current status of diagnosis and treatment of chronic lung allograft dysfunction / 器官移植

Chronic lung allograft dysfunction (CLAD) is the largest obstacle to the long-term survival of lung transplant recipients, which represents a series of complicated clinical manifestations of significant and persistent deterioration of lung allograft function after surgery. Due to lack of effective strategies for early diagnosis and prevention, over half of lung transplant recipients will develop CLAD within postoperative 5 years, which is likely to increase to 75% within postoperative 10 years. At present, no drug can be administered to completely prevent or reverse the progression of CLAD. In recent years, since the definition, diagnosis and treatment of CLAD have been updated by International Society of Heart and Lung Transplantation (ISHLT) in 2019, the understanding of CLAD has been significantly deepened within the international community. In this article, comprehensive diagnostic methods and potential treatment strategies of CLAD were explicitly illustrated, aiming to provide theoretical reference and insights for early monitoring and management of the incidence and progression of CLAD.

Pharmacological treatment of pulmonary fibrosis

Idiopathic pulmonary fibrosis (IPF) is a condition that has been described as alveolar collapse and thickening, which correlate with dysregulated surfactant production and injury to type 2 alveolar cells. As resolution of chest computed tomography has improved, especially with the development of high-resolution computed tomography (HRCT), the diagnostic measures adopted for pulmonary fibrosis has gradually shifted from biopsy to HRCT. This shift towards HRCT has aided in diagnostic evaluation and detection of the therapeutic and adverse effects of drugs for pulmonary fibrosis. Further, after the endpoint was changed to forced vital capacity, significant improvements are being observed in clinical trial outcomes. Currently active clinical trials are replacing lung biopsy with HRCT. In 2014, pirfenidone and nintedanib gained approval for tandem use in patients with IPF. These drugs were found to not only reduce the progression of pulmonary fibrosis, but also the acute exacerbation and mortality associated with the condition. These drugs showed consistent benefits regardless of the severity of patients' symptoms. Additionally, both nintedanib and pirfenidone were found to be effective in patients with advanced pulmonary fibrosis that was not classified as IPF. Nintedanib has been shown to reduce forced vital capacity in interstitial lung diseases associated with systemic sclerosis. In the next three to five years, many changes in treatment are expected, not only for IPF, but also for the entire spectrum of pulmonary fibrotic diseases. Pirfenidone and nintedanib are now considered standard treatments for IPF and few other fibrotic lung diseases. Clinicians treating patients with pulmonary fibrosis should keep themselves updated with the results of clinical trials that are currently underway.

Research Progress of Pirfenidone for connective tissue disease-associated interstitial lung diseases / 中国临床药理学与治疗学

Pirfenidone is an anti-fibrotic drug that has been shown to reduce the decline of lung function in patients with idiopathic pulmonary fibrosis (IPF) in multiple clinical trials, and has become first-line drugs for the treatment of IPF. Similar to the pathogenesis of IPF, clinical incidence of connective tissue disease-associated interstitial lung diseases (CTD-ILD) is high, and the clinical performance is not obvious. ILD may be the only or the original manifestation. Thus the clinical heterogeneity and the misdiagnosis rate are relatively high. Also, CTD-ILD is one of the important reasons that resulted in the death of patients. Currently, the clinical diagnosis and treatment of CTD-ILD still lack effective guide or unified agreement. Studies have shown that anti fibrosis drug pirfenidone shows some potential in the treatment of CTD-ILD. Here we summarize the research advances of pirfenidone in the treatment of CTD-ILD.

Progress in randomized clinical trials of antifibrotic drug nintedanib and pirfenidone / 中华劳动卫生职业病杂志

Pulmonary fibrosis is the terminal manifestation of a variety of interstitial lung diseases. Idiopathic pulmonary fibrosis (IPF) is one of the chronic, progressive, fibrotic lung disease with high incidence and poor prognosis. Nintedanib and pirfenidone are currently marketed anti-pulmonary fibrosis drugs, and their efficacy and safety are recognized in patients with IPF. This article reviews the targets and clinical trials of the two drugs, and provides a basis for the expansion of indications for anti-pulmonary fibrosis drugs.

Pirfenidone and nintedanib inhibit the lung alveolar regeneration in chronic pulmonary fibrosis mouse / 药学学报

The aim of this study was to determine whether the anti-fibrotic effects of pirfenidone (Pirf) and nintedanib (Nint) associated with the regulation of the alveolar epithelial type 2 cell (AEC II)-mediated lung alveolar regeneration in single- and multiple-dosage animal models of bleomycin-induced pulmonary fibrosis. All procedures involving animal treatment were approved according to the Committee on the Ethics of Animal Experiments of the Institute of Materia Medica, Chinese Academy of Medical Sciences. We found that the Pirf and Nint treatment of mice decreased the lung weight index, inflammation level, and the content of hydroxyproline compared with nontreated fibrotic mice in the single dosage model. Also, Pirf and Nint increased the oxygen saturation level and improved the lung functions in fibrotic mice, indicating that both drugs have anti-fibrotic effects in this model. However, the anti-fibrotic effects of Pirf and Nint were not observed in the multiple-dosage model. Further studies showed that Pirf and Nint decreased the expression of β-catenin, Axin2, c-Myc, Cyclin D1, and inhibited the Wnt/β-catenin signaling pathway, suggesting that Pirf and Nint did not produce anti-fibrotic effects in the multiple-dosage model due to their inhibiting the Wnt/β-catenin pathway and suppressing the stemness of AEC II, namely, suppressing AEC II-mediated lung alveolar regeneration.

Tratamiento farmacológico en la fibrosis pulmonar idiopática: drogas modificadoras de enfermedad / Pharmacological treatment in the FPI: disease modifying drugs

La historia natural de la fibrosis pulmonar idiopática (FPI) es heterogénea e impredecible. Aunque el curso de la enfermedad, sin tratamiento, es inevitablemente progresiva y de mal pronóstico. Los tratamientos históricos han variado desde corticosteroides e inmunosupresores (azatioprina, ciclofosfamida), hasta colchicina y N-acetilcisteína. En las últimas décadas se han realizado múltiples ensayos terapéuticos fallidos. Sin embargo, desde el año 2014 en los Estados Unidos, Europa y otros países, dos drogas, denominadas terapia antifibrótica o modificadoras de la enfermedad, están aprobadas para el tratamiento de la FPI: nintedanib y pirfenidona. La terapia antifibrótica, tiene como objetivo enlentecer en hasta 50% la declinación de la función pulmonar en pacientes con FPI.

The natural history of idiopathic pulmonary fibrosis (IPF) is heterogeneous and unpredictable. The course of the disease without treatment, is inevitably progressive, with a poor prognosis. Historical treatments have varied from corticosteroids and immunosuppressants (azathioprine, cyclophosphamide), to colchicine and N-acetylcysteine. In the last decades, multiple failed therapeutic trials have been carried out. However, since 2014 in the United States, Europe and other countries, two drugs, called antifibrotic therapy or disease modifiers, are approved for the treatment of IPF: nintedanib and pirfenidone. The purpose of antifibrotic therapy is to slow down the decline in lung function in patients with IPF up to 50%.

A brief review on Pleiotropic effects of Pirfenidone - novel and ongoing outcomes

Presently, there is a huge hype and excitement in the field of synthetic biologics and engineering regarding growing cases of their implication in various fields including health care systems. Furthermore, despite its large body of suggestive and fascinating accomplishments, the synthetic area is always been subject of much more prejudice and debate. However, over a couple of years, the generation of researchers had tested one of such disease modifying compound Pirfenidone (Esbriet®), to unleash their potential in the different disciplines of interventional pharmacology and therapy. In this pipeline, depending on its success of multiple missions, in context to advancing the therapy for different diseases, it became the first prescribed medicine to treat the people with one characteristic lung disorder called idiopathic pulmonary fibrosis (IPF). This review discusses the different therapeutic strategies beyond its well-known anti-fibrotic activity in several well-characterized animals, cell-based and human models and also regarding facts of Pirfenidone (PFD) as anti-inflammatory, anti-fibrinogenic, anti-oxidants including in the treatment of diabetic neuropathy, liver cirrhosis etc. This review also contains current investigations, focusing mainly on the novel findings and their outcomes in improving the quality of life of patients with different conditions and also suggests their implication on the basis of fundamental existential evidences to break the major impediment in transforming this disease-modifying drug into a personalized medicine.

Antifibrotic effect of pirfenidone on orbital fibroblasts in patients with thyroid-associated ophthalmopathy and its mechanisms / 中华内科杂志

Objective To investigate the effects of pirfenidone on orbital fibroblasts (OFs) from patients with thyroid-associated ophthalmopathy (TAO) and its underlying mechanisms.Methods OFs from patients with TAO were isolated and cultured in DMEM.Cells were divided into four groups and treated with 0,250,500 and 1 000 μg/ml pirfenidone for 24,48 or 72 hours,respectively.Cell proliferation was detected by tetramethyl azo salt (MTT) assay,and cell viability was determined by trypan blue.Transforming growth factor (TGF) β1 mRNA level was determined by real-time fluorescence quantitative PCR (RT-qPCR).Type Ⅰ and type 11Ⅲ collagen secreted from cultured cells were measured by enzyme-linked immuno sorbent assay (ELISA).Results (1) The primary cultured OFs had typical fibroblast spindle-like morphology.(2) MTT assay showed that pirfenidone treatment significantly inhibited the proliferation of OFs in a dose-dependent manner (P＜0.05) with the proliferation rates of pirfenidone treated groups of-15.31％,-24.92％,-48.53％ from 250,500,1 000 μg/ml after 72 h,respectively,in which the inhibition effect of 1 000 μg/ml pirfenidone was significantly different from the other two treated groups (P＜0.05).There were no significant differences in the inhibitory effect of the same concentration group among different time points at 24 h,48 h and 72 h (P＞0.05).Trypan blue showed that the survival rate of OFs in different concentrations of pirfenidone from 0,250,500,1 000 μ-g/ml at 72 h were 78.37％,79.21％,78.24％ and 76.28％,respectively.There were no significant differences between each drug treated and the control group (P＞0.05).(3) RT-qPCR results showed that the mRNA expression levels of TGFβ1 at 250,500,1 000 μg/ml pirfenidone treated groups at 72 h were 0.760±0.010,0.440±0.006,and 0.290±0.002,respectively.Compared with the control group (0.950±0.014),the differences were statistically significant (all P＜0.05).Moreover,TGFβ1 mRNA expression level in 1 000 μg/ml pirfenidone treated group was significantly lower than those in the other two treated groups (all P＜0.05).The secretion of type Ⅰ collagen (0.633 ± 0.006,0.527 ± 0.003 and 0.402±0.008) and type 11Ⅲ collagen (0.511±0.003,0.439±0.007 and 0.223±0.006) in 250,500 and 1 000 μg/ml pirfenidone treated groups at 72 h were significantly lower than those in the control group (0.794±0.005,0.527±0.007,all P＜0.05).Type Ⅰ and type Ⅲ collagen secretion in 1 000 μg/ml pirfenidone treated group were significantly lower than those in the other two groups (P＜0.05).Conclusions Pirfenidone inhibits the cell proliferation,TGFβ1 expression and collagen secretion of OFs,which may contribute to the anti-fibrotic effect of pirfenidone.

Pirfenidone inhibits TGF-β/Smad signaling pathway to alleviate hepatic cirrhosis induced by carbon tetrachloride / 中国免疫学杂志

Objective: This research aimed to explore the therapeutic effect and its mechanism of pirfenidone in liver cirrhosis induced by carbon tetrachloride in mice. Methods: Sixty male C57 BL/6 mice were randomly divided into the control group, model group and different doses of pirfenidone group, twelve rats in each group. Mice were intraperitoneally injected with 20％ CCl4 soybean oil solution ( 5 ml/kg), twice a week for 7 weeks. And these mice were free to drink 20％ ethanol solution in the third week after building the model. The low, medium and high dose groups were respectively given 50, 100 and 200 mg/kg of pirfenidone solution according to the body weights, while the model group and control group were given equal volume of blank solvent after building the model, once a day for 2 weeks. The serum level of ALT and AST, liver index, spleen index, the gene or protein expression level of TGF-β1 and Smad3 were analyzed before and after the treatment of pirfenidone. Results: The serum level of ALT, AST increased significantly in the model group ( P＜0. 05), while decreased significantly in different doses of pirfenidone group ( P＜0. 05). The liver and spleen index in the model group was significantly higher than that in the control group ( P＜0. 05). However, after treating with pirfenidone, the liver and spleen index were significantly lower than that in the model group ( P＜0. 05). The number of TGF-β1 positive cells in the model group was significantly more than that in the control group, but it was significantly decreased in the pirfenidone group. The gene expression level of Smad3 in the model group was significantly higher than that in the control group ( P＜0. 05). The gene expression level of TGF-β1 and Smad3 in different doses of pirfenidone group were significantly lower than that in the model group ( P＜ 0. 05). Meanwhile, the protein level of TGF-β1 and Smad3 were significantly increased in the model group, while decreased in the pirfenidone group. Conclusion: Pirfenidone relieves liver cirrhosis caused by carbon tetrachloride in mice by inhibiting the TGF-β1/Smad3 signaling pathway.

Research Progress of Two New Antifibrotic Drugs for Idiopathic Pulmonary Fibrosis / 中国药学杂志

Pirfenidone and nintadanib are two new anti-fibrotic drugs that have been shown to reduce the decline of lung function in patients with idiopathic pulmonary fibrosis (IPF) in multiple clinical trials,and have become first-line drugs for the treatment of IPF.However there are lack of references for how to choose between these two drugs. This article collected the latest research literatures related to pirfenidone or nintedanib,and summarized pharmacokinetics, pharmacological mechanism, clinical trials and safety profile of pirfenidone and nintedanib, so as to provide reference for clinical drug use. Evidences show both pirfenidone and nintedanib can reduce the decline of lung function and delay disease progression in mild-to-moderte IPF patients, and may have similar effects on severe IPF patients. There are large differences in pharmacological mechanisms, pharmacokinetics, and adverse events between these two new antifibrotic drugs. However, in terms of efficacy, there is no clear evidence to show which drug is better. Clinicians choosing drug should base on pharmacokinetic characteristics and adverse events between these two drugs.

Comparison of in-vitro anti-fibrotic effects of pirfenidone and nintedanib / 中国药理学通报

Aim To compare the mechanism of action of currently marketed pulmonary fibrosis drugs at the cellular level by evaluating the inhibitory effects of pirfenidone and nintedanib on the proliferation, migration and activation of human embryonic lung fibroblast HFL1. Methods The inhibitory effects of pirfenidone and nintedanib on TGF-β1/PDGF-induced HFL1 proliferation, migration and activation were evaluation of by MTT, scratch test,Q-PCR and Western blot. Results MTT, scratch test results showed that both pirfenidone and nintedanib could inhibit TGF-β1-induced fibroblast proliferation and migration, in which nintedanib had a higher titer than pirfenidone. In anti-activation experiments, both pirfenidone and nintedanib inhibited the expression of fibroblast activation markers mRNA and protein, and both inhibited the phosphorylation of Smad3 and inhibited the activation of TGF-β/ Smad3 signaling pathway. Nintedanib had a stronger inhibitory effect on TGF-β/Smad3 signaling pathway, which exerted 51 % inhibitory rate on Smad3 phosphorylation compared with 13% inhibitory rate of pirfenidone. Conclusions Both pirfenidone and nintedanib can inhibit the proliferation, migration and activation of myofibroblasts, where nintedanib has a higher inhibitory potency than pirfenidone does, and the inhibition of TGF-β/Smad3 signaling pathway is stronger.

Effect of Food on the Pharmacokinetics of Pirfenidone Capsules in Healthy Volunteers / 中国药师

Objective: To compare the pharmacokinetics and bioavailability of pirfenidone in the fasted and fed states in healthy volunteers. Methods: An open-label, randomized crossover study was conducted in 12 healthy subjects. Food effects were examined by comparing pharmacokinetic data of pirfenidone after administration of a single oral 400 mg dose under fasted or fed conditions. Plas-ma pirfenidone concentration was determined by an HPLC method and its pharmacokinetic parameters were calculated with DAS v2. 0 software. Results: Under fasted and fed conditions, the concentration-time profiles of pirfenidone were fitted a one-compartment model and the pharmacokinetic parameters were as follows: t1/2were (2. 16 ± 0. 47) and (2. 05 ± 0. 42) h;tmaxwere(0. 69 ± 0. 16)and (1. 46 ± 0. 40)h;Cmaxwere (12. 95 ± 1. 79) and (9. 16 ± 2. 87) mg·L-1;AUC0-12were (44. 97 ± 15. 06) and (36. 19 ± 14. 44) mg·h·L-1;AUC0-∞were (46. 55 ± 16. 79) and (37. 41 ± 15. 43) mg·h·L-1, respectively. When compared with that of the fasted group, tmaxwas significantly increased (P<0. 001) while Cmaxand AUC were remarkedly decreased in the fed group (P<0. 001 and P<0. 01, respectively). Conclusion: Concomitant food intake significantly influences the pharmacokinetics and bioavail-ability of pirfenidone as indicated by reducing its extent and rate of absorption, which is associated with better tolerability.

Pirfenidone inhibits TGF-β1-induced fibration in rat corneal stromal cells / 实用医学杂志

Objective To study the effect of pirfenidone (PFD) on the transformation of rat corneal stromal cells into fibroblasts in vitro and further explore the anti-fibrotic effect of PFD. Methods The corneal stromal cells from SD rat was isolated and cultured ,and was determined by vimentin stain. The experiment was divided into control group(DMEM+10%FBS),TGF-β1 group(2 ng/mL TGF-β1+DMEM+10%FBS)and PFD group(1 mg/mL PFD+ 2 ng/mL TGF-β1+DMEM+ 10%FBS). Cell proliferation was detected by CCK-8 assay. Collagen Ⅰ,Collagen Ⅲ,Keratocan and CD99 expression were detected by Western blot. Results Compared with control group and TGF-β1 group,the cell proliferation were significantly decreased in PFD group(P<0.05). Western blot showed that PFD can up-regulated Collagen Ⅰand Keratocan but down-regulated Collagen ⅢCD90 expression(P < 0.05). The ratio of Collagen Ⅲ/Collagen Ⅰ in PFD group was lowest in all groups(P < 0.05). Conclusion PFD can resistant fibration in corneal stromal cells may through the inhibition of TGF-β ,which affect the collagen synthesis and Keratocan,CD90 expression.