salvianolic-acid-B and Pulmonary-Fibrosis

Pulmonary fibrosis is the end‑stage manifestation of wide range of respiratory diseases and during pulmonary fibrosis, pulmonary inflammation and epithelial‑mesenchymal transition (EMT) play important roles. Salvianolic acid B (SAB) from the herb Salviae miltiorrhiza has been reported to possess an excellent anti‑inflammatory, antifibrotic and antioxidant activity. The present study aimed to investigate the ameliorative effect of SAB on bleomycin induced pulmonary fibrosis in mice. Adult albino mice were divided as SHAM/control group (saline alone), BLM group (bleomycin @ 1mg/kg intratracheally once) and SAB groups (BLM challenged once and SAB administration in three dosages @ 5, 10 and 15 mg/kg intraperitoneally daily for 30 days). Lungs wet/dry ratio and protein concentration in bronchoalveolar lavage fluid, MPO activity, oxidative stress markers, hydroxyproline assay, levels of inflammatory cytokines (TNF‑α, IL‑6 and TGF‑β1), NF‑κB activity, histopathology, immunostaining (E‑cadherin, vimentin and alpha ‑smooth muscle actin) and ultrastructural changes were studied. SAB showed anti‑inflammatory and anti‑fibrotic effects through inhibition of inflammatory cell infiltration, alveolar structure disruption, and collagen deposition and the expression of several fibrogenic cytokines. SAB also up‑regulate E‑cadherin and down‑regulated vimentin and alpha‑smooth muscle actin expression. In conclusion, Salvianolic acid B is effective in alleviating the BLM induced lung fibrosis through suppression of oxidative stress, inflammation, histological, ultrastructural changes and EMT.

Topics: Actins; Animals; Anti-Inflammatory Agents; Bleomycin; Cadherins; Cytokines; Mice; Pulmonary Fibrosis; Rodent Diseases; Vimentin

The purpose of this study was to investigate the antifibrotic effect and anticoagulant ability of salvianolic acid B (SAB) inhalation solution on bleomycin (BLM)-induced idiopathic pulmonary fibrosis (IPF) in rats. We investigated how the osmotic pressure and concentration of SAB in an aerosol exerted effects. We also determined the aerodynamic particle size distribution and the uniformity of the delivery dose; these parameters were found to be suitable for inhalation. Compared with BLM group, the levels of hydroxyproline (HYP), collagen-1 (Col-1), tissue factor (TF) / coagulation factor VII (TF-VIIa), activated coagulation factor X (FXa), thrombin-antithrombin complex (TAT), fibrinogen degradation product (FDP) and plasminogen activator inhibitor-1 (PAI-1) decreased in SAB group. The increased expression of coagulation factor Ⅱ (FⅡ), coagulation factor X (FX), tissue type plasminogen activator (t-PA) and urokinase type plasminogen activator (u-PA) proved that SAB has obvious antifibrotic and anticoagulant effects. Western blotting and immunofluorescence further showed that compared with the BLM group, the SAB group of rats exhibited significant reductions in the expression levels of protease-activated receptors-1 (PAR-1) and phospho-protein kinase C (p-PKC) and increased expression levels of protein kinase C (PKC) in lung tissue. Furthermore, SAB reduced the infiltration of lymphocytes and neutrophils, protected the basic structure of the lung from destruction, inhibited the proliferation of fibrous tissue. Collectively, our data revealed that SAB may exert its antifibrotic and anticoagulant effects by preventing the expression of PAR-1 and phosphorylation of PKC.

Topics: Administration, Inhalation; Animals; Anticoagulants; Antifibrinolytic Agents; Benzofurans; Bleomycin; Dose-Response Relationship, Drug; Drug Synergism; Drugs, Chinese Herbal; Male; Pulmonary Fibrosis; Rats; Rats, Sprague-Dawley

Pulmonary fibrosis (PF) is an interstitial lung disease characterized by interstitial inflammation and fibrosis. Salvianolic acid B (SAB) and sodium tanshinone IIA sulfonate (STS) are representative components in Salvia miltiorrhiza, which have been reported using in the treatment of PF. The aim of the study was to explain the role of inflammation in the process of PF and to investigate the effect of SAB and STS on inflammation and fibrosis in vitro. The results clearly indicated that lipopolysaccharide (LPS)-stimulated inflammatory response could induce fibroblast proliferation and fibroblast to myofibroblast transformation (FMT). Both SAB and STS significantly inhibited LPS-induced inflammation in vitro, including down-regulated the protein expression levels of IL-1β and TNF-α and the mRNA expression levels of IL1B and TNFA. Furthermore, both SAB and STS inhibited TGF-β1-induced the proliferation in MRC-5 cells and the overexpression of α-SMA and COL1α1, both the protein and mRNA levels. In conclusion, these results indicate that the inflammatory response is necessary for the development of PF, and the therapeutic effect of SAB and STS on PF may be related to anti-inflammatory and anti-fibrotic effects.

Topics: Actins; Anti-Inflammatory Agents; Benzofurans; Cell Proliferation; Coculture Techniques; Collagen Type I; Collagen Type I, alpha 1 Chain; Cytokines; Fibroblasts; Humans; Inflammation Mediators; Lung; Macrophages; Phenanthrenes; Pneumonia; Pulmonary Fibrosis; THP-1 Cells

Salvianolic acid B (SalB) is one of the most bioactive components extracted from Salvia miltiorrhiza, and its antioxidant capacity corresponds with its protective effects against cell injury from oxidative stress. The aim of the present study was to evaluate the effect of SalB on experimental pulmonary fibrosis and its ability to ameliorate the oxidative/antioxidative imbalance during fibrosis pathogenesis. The anti-fibrotic activity of SalB was first confirmed in Transforming growth factor β1(TGF-β1)-stimulated MRC-5 cells. The protection of SalB against oxidative stress during fibrogenesis in vitro was verified by detecting ROS production, the levels of glutathione (GSH) and malondialdehyde (MDA). The Western blot and PCR results indicated that SalB could up-regulate nuclear factor erythroid-derived 2-like 2 (Nrf2) at both the protein and mRNA levels and induce Nrf2 nuclear translocation in vitro, which may be the mechanism underlying the anti-fibrotic capacity of SalB. Furthermore, the anti-fibrotic and antioxidant capacities of SalB in vivo were confirmed in rats with BLM-induced pulmonary fibrosis. The immunohistochemistry results showed that Nrf2 was absent in fibroblastic foci (FF) areas, while the SalB treatment could increase the expression of Nrf2 in lung tissues, especially in FF areas.

Topics: Animals; Benzofurans; Cell Line; Cell Transdifferentiation; Dose-Response Relationship, Drug; Humans; Male; Myofibroblasts; NF-E2-Related Factor 2; Pulmonary Fibrosis; Rats; Rats, Wistar; Reactive Oxygen Species; Up-Regulation

Pulmonary fibrosis is a progressive and fatal disorder. In our previous study, we found that the Yiqihuoxue formula (YQHX), a prescription of Traditional Chinese Medicine, had a curative effect on scleroderma, a typical fibrotic disease. The aim of this study was to determine the key ingredient mediating the therapeutic effects of YQHX and to examine its effect on pulmonary fibrosis, including its mechanism. Luciferase reporter assays showed that the most important anti-fibrotic component of the YQHX was Salviae miltiorrhiza (SM). Experiments performed using a bleomycin-instilled mouse model of pulmonary fibrosis showed that Salvianolic acid B (SAB), the major ingredient of SM, had strong anti-inflammatory and anti-fibrotic effects through its inhibition of inflammatory cell infiltration, alveolar structure disruption, and collagen deposition. Furthermore, SAB suppressed TGF-β-induced myofibroblastic differentiation of MRC-5 fibroblasts and TGF-β-mediated epithelial-to-mesenchymal transition of A549 cells by inhibiting both Smad-dependent signaling and the Smad-independent MAPK pathway. Taken together, our results suggest that SM is the key anti-fibrotic component of the YQHX and that SAB, the major ingredient of SM, alleviates experimental pulmonary fibrosis both in vivo and in vitro by inhibiting the TGF-β signaling pathway. Together, these results suggest that SAB potently inhibits pulmonary fibrosis.

Topics: A549 Cells; Animals; Benzofurans; Bleomycin; Bronchoalveolar Lavage Fluid; Cell Differentiation; Cell Proliferation; Disease Models, Animal; Drugs, Chinese Herbal; Fibroblasts; Humans; Medicine, Chinese Traditional; Mice; NIH 3T3 Cells; Pulmonary Fibrosis; Salvia; Signal Transduction; Transforming Growth Factor beta

Salvianolic acid B (Sal B), one of the major water-soluble compounds of Danshen (a popular Chinese herb), possesses many of the biological activities, such as antifibrogenic effect in liver and renal diseases. Transforming growth factor-β1 (TGF-β1) plays a central role in the development of pulmonary fibrosis by stimulating extracellular matrix (ECM) accumulation and activating fibroblasts. Here, we investigated the effects of Sal B on cell proliferation, collagen synthesis, endogenous TGF-β1 production, and α-smooth muscle actin (α-SMA, a marker of myofibroblasts) expression in human lung fibroblasts stimulated by TGF-β1 in vitro. The cell proliferation rates were analyzed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assay. The expression of TGF-β1 and type I collagen at both the mRNA and protein levels was detected by reverse transcription polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and radioimmunoassay, respectively. The α-SMA expression was detected by Western blot. TGF-β1 treatment of lung fibroblasts increased cell proliferation rates, and enhanced the expression level of type I collagen, endogenous TGF-β1 production, and α-SMA expression (P < .05). The treatment with only Sal B did not affect the proliferation and differentiation of lung fibroblasts. Interestingly, Sal B was found to inhibit TGF-β1-induced cell proliferation, expression of type I collagen, endogenous TGF-β1 production, and α-SMA expression in lung fibroblasts. Moreover, the inhibitory effect of Sal B on TGF-β1-induced proliferation and differentiation in lung fibroblasts was more significant when treated with high-dose Sal B (1 μmol/L versus 10 μmol/L, P < .05). These data demonstrate that Sal B inhibits TGF-β1-induced cell proliferation and differentiation in vitro experiment.

Topics: Actins; Benzofurans; Cell Differentiation; Cell Line; Cell Proliferation; Collagen Type I; Down-Regulation; Drug Evaluation, Preclinical; Drugs, Chinese Herbal; Fibroblasts; Humans; Phytotherapy; Pulmonary Fibrosis; Salvia miltiorrhiza; Transforming Growth Factor beta1