Ginkgo biloba Extract EGb761 Attenuates Bleomycin-Induced Experimental Pulmonary Fibrosis in Mice by Regulating the Balance of M1/M2 Macrophages and Nuclear Factor Kappa B (NF-κB)-Mediated Cellular Apoptosis.

BACKGROUND The aim of this study was to show whether the standardized Ginkgo biloba extract EGb761, a traditional Chinese medicine, has a therapeutic effect on pulmonary fibrosis (PF). MATERIAL AND METHODS Bleomycin (BLM) was used for establishing the PF mouse model. The mice were treated with a gradient of EGb761 for 28 days to determine an appropriate drug dose. On day 28, the effect of EGb761 on lung injury and inflammation was confirmed by hematoxylin and eosin and Masson staining and evaluated by pulmonary alveolitis and Ashcroft score. The balance of M1/M2 macrophages was evaluated with the respective markers inducible nitric oxide synthase and and interleukin-10 by real-time polymerase chain reaction. Furthermore, the expressions of fibrosis-associated protein alpha-smooth muscle actin (SMA), related inflammatory protein transforming growth factor (TGF)-ß1, the apoptosis-related proteins B-cell lymphoma-associated X protein (Bax), B-cell lymphoma (Bcl)-2, caspase-3, caspase-9, and phosphorylated nuclear factor (NF)-kappaB (p65) were assessed by western blot. RESULTS On day 28, PF was induced by treating with BLM, whereas EGb761 suppressed the PF of lung tissue. The BLM-induced imbalance of M1/M2 macrophages was reduced by EGb761. Furthermore, the increasing amounts of alpha-SMA and TGF-ß1 induced by BLM were suppressed by EGb761. In addition, the protein or messenger ribonucleic acid expression levels of phosphorylated NF-kappaB (p65), caspase-3, and caspase-9 were upregulated, whereas Bax and Bcl-2 were downregulated. Treatment with EGb761 restored the levels of these proteins except for caspase-9. CONCLUSIONS This study illustrated the protective effect of EGb761 on BLM-induced PF by regulating the balance of M1/M2 macrophages and NF-kappaB (p65)-mediated apoptosis. The results demonstrated the potential clinical therapeutic effect of EGb761, providing a novel possibility for curing PF.

Notch signaling pathway mediates the immunomodulatory mechanism of Yangfei Huoxue decoction alleviating bleomycin-induced pulmonary fibrosis in rats.

OBJECTIVE: To investigate the immunomodulatory mechanism by which Yangfei Huoxue decoction (YHD) alleviates bleomycin (BLM)-induced pulmonary fibrosis (PF) in rats. METHODS: Rats were randomly divided into two time-point groups (day 14 and 28), and each time-point group comprised the following six subgroups: control, BLM, dexamethasone (DXM), YHD high dose (YHD-H), YHD middle dose (YHD-M), and YHD low dose (YHD-L). Haematoxylin and eosin and Masson staining, flow cytometry, enzyme-linked immunosorbent assay, Western blotting and UPLC-QT of analyses were examined. RESULTS: The results showed that YHD reduced the degree of alveolar inflammation and fibrosis; downregulated the expression of CD28, CD80, CD86, Delta-like 1, Notch2, and Notch3; and upregulated the proportions of Th1/Th2 and Tc1/Tc2. The seven components of YHD were detected. CONCLUSION: The current study indicates that YHD mainly functions by regulating the immune system and that the molecular mechanism may be related to the regulation of the Notch signaling pathway.

Protective Effects of Functional Amino Acids on Apoptosis, Inflammatory Response, and Pulmonary Fibrosis in Lipopolysaccharide-Challenged Mice.

Lung injury is a complicated and lethal condition characterized by alveolar barrier disruption, pulmonary edema, enhanced inflammation, and apoptosis in alveoli. However, therapeutic strategies to ameliorate lung injury without exerting side effects are not available. Functional amino acids have been shown to have anti-inflammatory and anti-apoptotic effects under various conditions. The objective of this study was to test the hypothesis that arginine, glutamine, or glycine supplementation ameliorated lipopolysaccharide (LPS)-induced lung injury in mice. Mice pretreated with aerosolized arginine, glutamine, or glycine were exposed to aerosolized LPS to induce lung injury. Results showed that arginine or glycine pretreatment beneficially reduced LPS-induced collagen deposition, apoptosis of alveolar cells, expression of inflammatory cytokines and chemokines, and accumulation of neutrophils and macrophages in lung tissues of mice, thus contributing to improved alveolar integrity and function. Glutamine administration reduced LPS-induced collagen deposition and inflammatory cytokines without affecting any other parameters examined in the study. Our findings indicated that arginine or glycine pretreatment effectively alleviated LPS-induced lung injury by inhibiting the accumulation of lymphocytes, the release of inflammatory cytokines and chemokines, and the apoptosis of alveolar cells. Supplementation of arginine or glycine may be a novel nutritional strategy to reduce deleterious effects of bacterial infection on alveolar function.

Andrographolide ameliorates silica induced pulmonary fibrosis.

The purpose of this study was to investigate the protective effect of andrographolide in silica-induced pulmonary fibrosis (PF) in mice and its underlying mechanisms. Male Swiss albino mice were divided into five groups: Normal control group, disease control group (1.5 mg silica/60 µL/mice) via oropharyngeal route, low dose (LD) group received silica + andrographolide (3 mg/kg), high dose (HD) group received silica + andrographolide (10 mg/kg), andrographolide per se group received 10 mg/kg andrographolide. Various bronchoalveolar lavage fluid (BALF) and biochemical parameters, inflammatory cytokines, histology and protein expression studies were carried out. Andrographolide significantly reduced total protein concentration, albumin, accumulation of inflammatory cells and lactate dehydrogenase (LDH) level in BALF. We found that andrographolide intervention led to decreased levels of the inflammatory cells including neutrophils, macrophages and lymphocytes in the BALF of the treated animals. In addition, andrographolide significantly reduced nitrite (p < 0.01 at HD), malondialdehyde (p < 0.01 at HD) and upregulated glutathione (p < 0.01 at HD) in silica challenged animals. Andrographolide showed anti-fibrotic activity by reducing collagen deposition and inflammation in lung. Histopathology revealed that andrographolide decreased irregular cellular nodules, inflammatory infiltration and fibrosis. Andrographolide intervention significantly reduced the expression of N-cadherin, α-SMA and vimentin (mesenchymal markers) and upregulated the expression of E-cadherin (an epithelial marker). Hence, andrographolide elicits its anti-pulmonary fibrotic effect by halting the progression of epithelial-to-mesenchymal transition (EMT) via affecting fibroblasts. We, to the best of our knowledge prove for the first time that andrographolide possesses potent antifibrotic activity by targeting inflammatory cells and EMT associated fibroblasts.

Gentiopicroside ameliorates bleomycin-induced pulmonary fibrosis in mice via inhibiting inflammatory and fibrotic process.

Pulmonary fibrosis (PF) is a chronic and ultimately fatal interstitial lung disease of various causes. The advent of nintedanib and pirfenidone provides treatment options for PF patients for the first time. However, the adverse effects of the two drugs such as gastrointestinal disorders and hepatic dysfunction often lead to treatment discontinuation. Gentiopicroside (GPS) is a natural secoiridoid glycoside from gentian species of medicinal plants, and has a variety of pharmacological activities, including hepatoprotective and cholagogic, anti-inflammatory, antinociceptive, and smooth muscle relaxing activities. The present study aimed to investigate the therapeutical effects of GPS on bleomycin (BLM)-induced PF in mice. Severe lung inflammation and fibrosis were observed in BLM-treated mice. GPS significantly ameliorated inflammatory and fibrotic responses in lungs of PF mice which were confirmed by histopathological examinations including light microscopy and transmission electron microscopy. Additionally, GPS significantly decreased the levels of inflammatory cytokines including TNF-α and IL-1ß in bronchoalveolar lavage fluid and reduced the content of hydroxyproline in lungs of PF mice. Furthermore, GPS significantly downregulated the expression of TGF-ß1 and CTGF in lungs of PF mice. In vitro, GPS inhibited epithelial-mesenchymal transition of A549 cells stimulated by TGF-ß1, in a dose-dependent manner. Our findings suggest that GPS has the potential as an ideal drug candidate for PF, as it has both anti-inflammatory and anti-fibrotic effects. Alveolar epithelial cells and TGF-ß1 may be the main target cells and molecule of GPS on BLM-induced PF, respectively.

Piper nigrum extract ameliorated allergic inflammation through inhibiting Th2/Th17 responses and mast cells activation.

Piper nigrum (Piperaceae) is commonly used as a spice and traditional medicine in many countries. P. nigrum has been reported to have anti-oxidant, anti-bacterial, anti-tumor, anti-mutagenic, anti-diabetic, and anti-inflammatory properties. However, the effect of P. nigrum on allergic asthma has not been known. This study investigated the effect of P. nigrum ethanol extracts (PNE) on airway inflammation in asthmatic mice model. In the ovalbumin (OVA)-induced allergic asthma model, we analysed the number of inflammatory cells and cytokines production in bronchoalveolar lavage fluid (BALF) and lung tissue; histological structure; as well as the total immunoglobulin (Ig)E, anti-OVA IgE, anti-OVA IgG1 and histamine levels in serum. The oral administration (200 mg/kg) of PNE reduced the accumulation of inflammatory cells (eosinophils, neutrophils in BALF and mast cells in lung tissue); regulated the balance of the cytokines production of Th1, Th2, Th17 and Treg cells, specifically, inhibited the expressions of GATA3, IL-4, IL-6, IL-1ß, RORγt, IL-17A, TNF-α and increased the secretions of IL-10, INF-γ in BALF and lung homogenate. Moreover, PNE suppressed the levels of total IgE, anti-OVA IgE, anti-OVA IgG1 and histamine release in serum. The histological analysis showed that the fibrosis and infiltration of inflammatory cells were also ameliorated in PNE treated mice. On the other hand, PNE inhibited the allergic responses via inactivation of rat peritoneal mast cells degranulation. These results suggest that PNE has therapeutic potential for treating allergic asthma through inhibiting Th2/Th17 responses and mast cells activation.

Grape seed extract ameliorates bleomycin-induced mouse pulmonary fibrosis.

Pulmonary fibrosis is common in a variety of inflammatory lung diseases, such as interstitial pneumonia, chronic obstructive pulmonary disease, and silicosis. There is currently no effective clinical drug treatment. It has been reported that grape seed extracts (GSE) has extensive pharmacological effects with minimal toxicity. Although it has been found that GSE can improve the lung collagen deposition and fibrosis pathology induced by bleomycin in rat, its effects on pulmonary function, inflammation, growth factors, matrix metalloproteinases and epithelial-mesenchymal transition remain to be researched. In the present study, we studied whether GSE provided protection against bleomycin (BLM)-induced mouse pulmonary fibrosis. ICR strain mice were treated with BLM in order to establish pulmonary fibrosis models. GSE was given daily via intragastric administration for three weeks starting at one day after intratracheal instillation. GSE at 50 or 100mg/kg significantly reduced BLM-induced inflammatory cells infiltration, proinflammatory factor protein expression, and hydroxyproline in lung tissues, and improved pulmonary function in mice. Additionally, treatment with GSE also significantly impaired BLM-induced increases in lung fibrotic marker expression (collagen type I alpha 1 and fibronectin 1) and decreases in an anti-fibrotic marker (E-cadherin). Further investigation indicated that the possible molecular targets of GSE are matrix metalloproteinases-9 (MMP-9) and TGF-ß1, given that treatment with GSE significantly prevented BLM-induced increases in MMP-9 and TGF-ß1 expression in the lungs. Together, these results suggest that supplementation with GSE may improve the quality of life of lung fibrosis patients by inhibiting MMP-9 and TGF-ß1 expression in the lungs.

Earthworm extract attenuates silica-induced pulmonary fibrosis through Nrf2-dependent mechanisms.

Silicosis is an occupational pulmonary fibrosis caused by inhalation of silica (SiO2) and there are no ideal drugs to treat this disease. Earthworm extract (EE), a natural nutrient, has been reported to have anti-inflammatory, antioxidant, and anti-apoptosis effects. The purpose of the current study was to test the protective effects of EE against SiO2-induced pulmonary fibrosis and to explore the underlying mechanisms using both in vivo and in vitro models. We found that treatment with EE significantly reduced lung inflammation and fibrosis and improved lung structure and function in SiO2-instilled mice. Further mechanistic investigations revealed that EE administration markedly inhibited SiO2-induced oxidative stress, mitochondrial apoptotic pathway, and epithelial-mesenchymal transition in HBE and A549 cells. Furthermore, we demonstrate that Nrf2 activation partly mediates the interventional effects of EE against SiO2-induced pulmonary fibrosis. Our study has identified EE to be a potential anti-oxidative, anti-inflammatory, and anti-fibrotic drug for silicosis.

Salidroside protects against bleomycin-induced pulmonary fibrosis: activation of Nrf2-antioxidant signaling, and inhibition of NF-κB and TGF-ß1/Smad-2/-3 pathways.

Pulmonary fibrosis (PF) can severely disrupt lung function, leading to fatal consequences. Salidroside is a principal active ingredient of Rhodiola rosea and has recently been reported to protect against lung injures. The present study was aimed at exploring its therapeutic effects on PF. Lung fibrotic injuries were induced in SD rats by a single intratracheal instillation of 5 mg/kg bleomycin (BLM). Then, these rats were administrated with 50, 100, or 200 mg/kg salidroside for 28 days. BLM-triggered structure distortion, collagen overproduction, excessive inflammatory infiltration, and pro-inflammatory cytokine release, and oxidative stress damages in lung tissues were attenuated by salidroside in a dose-dependent manner. Furthermore, salidroside was noted to inhibit IκBα phosphorylation and nuclear factor kappa B (NF-κB) p65 nuclear accumulation while activating Nrf2-antioxidant signaling in BLM-treated lungs. Downregulation of E-cadherin and upregulation of vimentin, fibronectin, and α-smooth muscle actin (α-SMA) indicated an epithelial-mesenchymal transition (EMT)-like shift in BLM-treated lungs. These changes were suppressed by salidroside. The expression of TGF-ß1 and the phosphorylation of its downstream targets, Smad-2/-3, were enhanced by BLM, but weakened by salidroside. Additionally, salidroside was capable of reversing the recombinant TGF-ß1-induced EMT-like changes in alveolar epithelial cells in vitro. Our study reveals that salidroside's protective effects against fibrotic lung injuries are correlated to its anti-inflammatory, antioxidative, and antifibrotic properties.

Pyunkang-hwan (Pyunkang-Tang) ameliorates air pollutant-induced inflammatory hypersecretion of airway mucus and bleomycin- induced pulmonary fibrosis in rats.

OBJECTIVE: To investigate the effect of Pyunkanghwan (Pyunkang-tang) extract (PGT) on secretion of airway mucin in an experimental animal model involving hyperplasia of goblet cells and mucus hypersecretion, and to test its effects on bleomycin (BLM)-induced pulmonary fibrosis in vivo. METHODS: The protective activity of orally administered PGT was assessed in two rat pulmonary disease models. Effects on hypersecretion of pulmonary mucin in sulfur dioxide (SO2)-induced bronchitis in rats were assessed by quantifying the amount of mucus secreted and examining histopathology in the tracheal epithelium. In a rat model for BLMinduced pulmonary fibrosis, toxicity to the pulmonary system was examined by measuring levels of malondialdehyde and hydroxyproline, indicators of lipid peroxides and collagen, respectively, in lung tissue 28 days post-BLM treatment. Serial sections of lung tissue were stained with Masson trichrome to visualize collagen deposition. Effects of PGT on collagen synthesis were also assessed in vitro, in a cell culture model. RESULTS: PGT inhibited mucin secretion and normalized SO2-induced increased muco- substances in goblet cells. In the BLM- induced model, PGT decreased the characteristic histopathological features of lung fibrosis and inhibited fibrotic lesions, as indicated by decreased hydroxyproline content. PGT also inhibited the BLM-induced increase in malondialdehyde levels, demonstrating its protective effect against lipid peroxidation in cell membranes of the lung. In MLg 2908 mouse lung fibroblast cells, PGT decreased transforming growth factor (TGF)-ß-stimulated type I collagen synthesis. CONCLUSION: PGT can inhibit both hypersecretion of airway mucins and pulmonary fibrosis.

Protective effect of polyphenols in an inflammatory process associated with experimental pulmonary fibrosis in mice.

Polyphenols have been described to have a wide range of biological activities, and many reports, published during recent years, have highlighted the beneficial effects of phenolic compounds, illustrating their promising role as therapeutic tools in several acute and chronic disorders. The purpose of study was to evaluate, in an already-assessed model of lung injury caused by bleomycin (BLM) administration, the role of resveratrol and quercetin, as well as to explore the potential beneficial properties of a mango leaf extract, rich in mangiferin, and a grape leaf extract, rich in dihydroquercetin (DHQ), on the same model. Mice were subjected to intra-tracheal administration of BLM, and polyphenols were administered by oral route immediately after BLM instillation and daily for 7 d. Treatment with resveratrol, mangiferin, quercetin and DHQ inhibited oedema formation and body weight loss, as well as ameliorated polymorphonuclear infiltration into the lung tissue and reduced the number of inflammatory cells in bronchoalveolar lavage fluid. Moreover, polyphenols suppressed inducible nitric oxide synthase expression, and prevented oxidative and nitroxidative lung injury, as shown by the reduced nitrotyrosine and poly (ADP-ribose) polymerase levels. The degree of apoptosis, as evaluated by Bid and Bcl-2 balance, was also suppressed after polyphenol treatment. Finally, these natural products down-regulated cyclo-oxygenase-2, extracellular signal-regulated kinase phosphorylated expression and reduced NF-κBp65 translocation. Our findings confirmed the anti-inflammatory effects of resveratrol and quercetin in BLM-induced lung damage, and highlight, for the first time, the protective properties of exogenous administration of mangiferin and DHQ on experimental pulmonary fibrosis.

Acute and subacute pulmonary toxicity and mortality in mice after intratracheal instillation of ZnO nanoparticles in three laboratories.

Inhalation is the main pathway of ZnO exposure in the occupational environment but only few studies have addressed toxic effects after pulmonary exposure to ZnO nanoparticles (NP). Here we present results from three studies of pulmonary exposure and toxicity of ZnO NP in mice. The studies were prematurely terminated because interim results unexpectedly showed severe pulmonary toxicity. High bolus doses of ZnO NP (25 up to 100 µg; ≥1.4 mg/kg) were clearly associated with a dose dependent mortality in the mice. Lower doses (≥6 µg; ≥0.3 mg/kg) elicited acute toxicity in terms of reduced weight gain, desquamation of epithelial cells with concomitantly increased barrier permeability of the alveolar/blood as well as DNA damage. Oxidative stress was shown via a strong increase in lipid peroxidation and reduced glutathione in the pulmonary tissue. Two months post-exposure revealed no obvious toxicity for 12.5 and 25 µg on a range of parameters. However, mice that survived a high dose (50 µg; 2.7 mg/kg) had an increased pulmonary collagen accumulation (fibrosis) at a similar level as a high bolus dose of crystalline silica. The recovery from these toxicological effects appeared dose-dependent. The results indicate that alveolar deposition of ZnO NP may cause significant adverse health effects.

Ojeok-san, a traditional Korean herbal medicine attenuates airway inflammation and pulmonary fibrosis induced by repeated ovalbumin challenge.

ETHNOPHARMACOLOGICAL EVIDENCE: Ojeok-san, a traditional Korean herbal medicine, is widely used in China, Japan and Korea for treatment of the common cold, pain and fever. AIM OF THE STUDY: In this study, we investigated the protective effects of Ojeok-san aqueous extract (OJS) against pulmonary fibrosis using a chronic asthma murine model. METHODS: Mice were sensitized by intraperitoneal injection of ovalbumin (OVA), followed 1 weeks later by an airway challenge with OVA delivered three times a week for 4 weeks. OJS (50mg/kg or 100mg/kg) was also administered by oral gavage once a day for 4 weeks. RESULTS: OJS significantly reduced interleukin (IL)-4, IL-13, eotaxin, immunoglobulin E and the number of inflammatory cells in bronchoalveolar lavage fluid; in addition, it reduced inflammatory cell infiltration and mucus production in the respiratory tract. We found that OJS also attenuated the OVA-induced increase in vascular endothelial growth factor (VEGF), transforming growth factor (TGF)-ß1 and Smad3 protein in lung tissue, as determined by Western analysis and immunohistochemistry. These inhibitory effects of OJS were accompanied by a reduction in pulmonary fibrosis, consistent with the histopathology of lung tissue stained with Masson's trichrome. CONCLUSION: Administration of OJS reduced the airway inflammation and pulmonary fibrosis, as well as the level of T helper type 2 cytokines and VEGF and TGF-ß1/Smad3 expressions in lung tissue. These results suggest that OJS might represent a useful new oral therapy for the treatment of chronic asthma.

High-dose N-acetylcysteine decreases silica-induced lung fibrosis in the rat.

OBJECTIVE: To study the potential of high-dose N-acetylcysteine (NAC) to attenuate silica-induced pulmonary fibrosis in the rat. METHODS: Rats exposed to intratracheal instillation of silica particles were treated with 500 mg/kg NAC orally every day for 7 days, before and up to 28 days after silica administration (n = 32), or received no treatment following silica exposure (n = 32); a third group received intratracheal saline (n = 32). Fibrosis score, and hydroxyproline (HYP) and malondialdehyde (MDA) content, were assessed in lung tissue. Bronchoalveolar lavage fluid (BALF) and serum levels of tumour necrosis factor (TNF)-α, interleukin (IL)-8 and high-sensitivity C-reactive protein (hsCRP) were assessed by enzyme-linked immunosorbent assay. RESULTS: Histopathology revealed inflammation and fibrosis in lung tissue from rats exposed to silica, but not in saline controls. The fibrosis score was significantly lower in animals treated with NAC compared with silica-exposed untreated rats. HYP and MDA content were significantly lower at all timepoints, following NAC treatment versus no treatment, in silica-exposed rats. NAC attenuated silica-induced increases in TNF-α, IL-8 and hsCRP in BALF and serum. CONCLUSIONS: Oral treatment with high-dose NAC during early silica exposure can ameliorate the activity of proinflammatory cytokines, thus attenuating subsequent lung fibrosis. These results suggest that NAC has potential as a treatment for silica-induced lung fibrosis.

Protective effects of neferine on amiodarone-induced pulmonary fibrosis in mice.

The effects of neferine, a bisbenzylisoquinline alkaloid extracted from the Chinese traditional medicine seed embryo of Nelumbo nucifera Gaertn, on amiodarone-induced pulmonary fibrosis in mice were evaluated. Adult Kunming mice were induced to develop pulmonary fibrosis through intratracheal instillation of amiodarone (6.25 mg/kg) on the 1st, 3rd and 5th day. Mice were treated orally with saline, neferine (20 mg/kg), prednisolone (15 mg/kg), pirfenidone (100 mg/kg) twice a day after the third amiodarone instillation. On Day 21, all the lung tissues were collected for hydroxyproline measurement and the histological examination by hematoxylin-eosin and Masson staining. All the blood sample were collected for surfactant protein-D (SP-D) levels assay, Th1/Th2 balance valuation, CD4+CD25+ regulatory T cells (Tregs) analysis by Enzyme-linked immunosorbent assay and flow cytometry. Our data showed that neferine significantly restored the significant reductions in body weights, the increased levels of lung index and hydroxyproline, the abnormal histological findings, the serum SP-D increase, the Th1/Th2 imbalance by decreasing IL-4 and increasing IFN-γ levels and the increases in the population of CD4+CD25+ Tregs associated with amiodarone instillation in mice. Similar changes were also observed in the prednisolone or pirfenidone treated mice. In conclusion, these results indicated that neferine possessed a significant inhibitory effect on amiodarone-induced pulmonary fibrosis, probably due to its properties of anti-inflammation, SP-D inhibition and restoring increased CD4+CD25+ Tregs which may modulate Th1/Th2 imbalance by suppressing Th2 response (from Th2 polarity toward a Th1 dominant response).

Modulatory effects of curcumin and green tea extract against experimentally induced pulmonary fibrosis: a comparison with N-acetyl cysteine.

The study was aimed to investigate the protective effect of green tea extract (GTE), curcumin, and N-acetyl cysteine (NAC) on experimentally induced pulmonary fibrosis. Curcumin (200 mg/kg b.w), GTE (150 mg/kg b.w), and NAC (490 mg/kg b.w) were administered orally for 14 days with concomitant administration of cyclophosphamide (CP). Lung fibrosis was assessed by measuring hydroxyproline and elastin levels and confirmed by histopathological examination. Oxidative stress was also observed in the CP group. Lung myeloperoxidase activity was significantly decreased in animals of the CP group. N-acetyl-ß-d-glucosaminidase, leukotriene C4, and protein were increased in bronchoalveolar lavage fluid (BALF). Transforming growth factor-ß, interleukin -1ß, and histamine were increased in both serum and BALF. All modulators markedly attenuated the altered biochemical parameters as compared to CP-treated rats. These results suggest the possibility of using these treatments as protective agents with chemotherapy and as protective agents for lung fibrosis.

Fasudil, a Rho-kinase inhibitor, attenuates bleomycin-induced pulmonary fibrosis in mice.

The mechanisms underlying the pathogenesis of idiopathic pulmonary fibrosis (IPF) involve multiple pathways, such as inflammation, epithelial mesenchymal transition, coagulation, oxidative stress, and developmental processes. The small GTPase, RhoA, and its target protein, Rho-kinase (ROCK), may interact with other signaling pathways known to contribute to pulmonary fibrosis. This study aimed to determine the beneficial effects and mechanisms of fasudil, a selective ROCK inhibitor, on bleomycin-induced pulmonary fibrosis in mice. Our results showed that the Aschcroft score and hydroxyproline content of the bleomycin-treated mouse lung decreased in response to fasudil treatment. The number of infiltrated inflammatory cells in the bronchoalveolar lavage fluid (BALF) was attenuated by fasudil. In addition, fasudil reduced the production of transforming growth factor-ß1 (TGF-ß1), connective tissue growth factor (CTGF), alpha-smooth muscle actin (α-SMA), and plasminogen activator inhibitor-1 (PAI-1) mRNA and protein expression in bleomycin-induced pulmonary fibrosis. These findings suggest that fasudil may be a potential therapeutic candidate for the treatment of pulmonary fibrosis.

Sumas Mountain chrysotile induces greater lung fibrosis in Fischer344 rats than Libby amphibole, El Dorado tremolite, and Ontario ferroactinolite.

The physical properties of different types of asbestos may strongly affect health outcomes in exposed individuals. This study was designed to provide understanding of the comparative toxicity of naturally occurring asbestos (NOA) fibers including Libby amphibole (LA), Sumas Mountain chrysotile (SM), El Dorado tremolite (ED), and Ontario ferroactinolite (ON) cleavage fragments. Rat-respirable fractions (PM2.5) were prepared by water elutriation. Surface area was greater for SM (64.1 m²/g) than all other samples (range: 14.1-16.2 m²/g), whereas mean lengths and aspect ratios (ARs) for LA and SM were comparable and greater than ED and ON. Samples were delivered via a single intratracheal (IT) instillation at doses of 0.5 and 1.5mg/rat. One day post-IT instillation, low-dose NOA exposure resulted in a 3- to 4-fold increase in bronchoalveolar lavage fluid (BALF) cellularity compared with dispersion media (DM) controls, whereas high-dose exposure had a more severe effect on lung inflammation which varied by source. Although inducing less neutrophilic inflammation than ON and ED, exposure to either LA or SM resulted in a greater degree of acute lung injury. Three months post-IT instillation, most BALF parameters had returned to control levels, whereas the development of fibrosis persisted and was greatest in SM-exposed rats (SM > LA > ON > ED). These data demonstrate that fiber length and higher AR are directly correlated with the severity of fibrosis and that, in the rat, exposure to SM is more fibrogenic than LA which suggests that there may be cause for concern for people at risk of being exposed to NOA from the Sumas Mountain landslide.

Houttuynia cordata inhibits lipopolysaccharide-induced rapid pulmonary fibrosis by up-regulating IFN-γ and inhibiting the TGF-ß1/Smad pathway.

This study aimed to explore the effect and mechanism of H. cordata vapor extract on acute lung injury (ALI) and rapid pulmonary fibrosis (RPF). We applied the volatile extract of HC to an RPF rat model and analyzed the effect on ALI and RPF using hematoxylin-eosin (H&E) staining, routine blood tests, a cell count of bronchoalveolar lavage fluid (BALF), lactate dehydrogenase (LDH) content, van Gieson (VG) staining, hydroxyproline (Hyp) content and the dry/wet weight ratio. The expression of IFN-γ/STAT(1), IL-4/STAT(6) and TGF-ß(1)/Smads was analyzed using ELISA, immunohistochemistry and western blotting methods. The active ingredients of the HC vapor extract were analyzed using a gas chromatograph-mass spectrometer (GC-MS), and the effects of the active ingredients of HC on the viability of NIH/3T3 and RAW264.7 cells were detected using an MTT assay. The active ingredients of the HC vapor extract included 4-terpineol, α-terpineol, l-bornyl acetate and methyl-n-nonyl ketone. The results of the lung H&E staining, Hyp content, dry/wet weight ratio and VG staining suggested that the HC vapor extract repaired lung injury and reduced RPF in a dose-dependent manner and up-regulated IFN-γ and inhibited the TGF-ß1/Smad pathway in vivo. In vitro, it could inhibit the viability of RAW264.7 and NIH/3T3 cells. It also dose-dependently inhibited the expression of TGF-ß1 and enhanced the expression of IFN-γ in NIH/3T3. The HC vapor extract inhibited LPS-induced RPF by up-regulating IFN-γ and inhibiting the TGF-ß1/Smad pathway.