Radix puerariae extracts ameliorate paraquat-induced pulmonary fibrosis by attenuating follistatin-like 1 and nuclear factor erythroid 2p45-related factor-2 signalling pathways through downregulation of miRNA-21 expression.

BACKGROUND: Puerarin, extracted from Radix puerariae, was reported to ameliorate airway inflammation, lung injury and lung fibrosis induced by paraquat (PQ) in mice. However, effects of Radix puerariae extracts (RPEs) on lung fibrosis or signalling pathways in PQ-induced lung injury have not been well studied. Therefore, the goals of our study were to investigate whether Radix puerariae extracts are antifibrotic in a paraquat (PQ) induced lung fibrosis model in mice and to propose possible mechanisms of action of the RPE effects. METHODS: We used a long-term exposure model of PQ-induced lung fibrosis in mice to evaluate effects of antioxidant-containing RPE. We examined effects of miR-21 on follistatin-like 1 (Fstl 1) pathways and oxidative stress in the lung. Gene expression levels of miR-21, Fstl 1, transforming growth factor-ß1 (TGF-ß1), connective tissue growth factor (CTGF), collagen-1 and collagen III were measured by real-time PCR. Protein expression levels of Fstl 1(FSTL1), heme oxygenase-1 (HO-1), nuclear factor erythroid 2p45-related factor-2 (Nrf2), Smad2/3, p38MAPK, nuclear factor-κB 65 (NF-κB65), and matrix metalloproteinase-9 were detected by western blotting. FSTL1 andalpha-smooth muscle actin (α-SMA) in lung tissue were detected by immunohistochemistry. Malondialdehyde, superoxide dismutase (SOD), reduced (GSH) and oxidised (GSSH) glutathione and reactive oxygen species levels, hydroxyproline and total lung collagen were also determined. RESULTS: Long-term challenge with PQ enhanced miRNA-21 (miR-21), Fstl 1 pathways, oxidative stress and development of fibrotic features in the lungs. RPE reduced features of lung fibrosis by blocking Fstl 1 pathways and oxidative stress through decreased miR-21 expression. This was accompanied by suppression of CTGF, TGF-ß1, vascular endothelial growth factor, collagen I, and collagen III. In addition, PQ-induced activation of NF-κB, Nrf2 and α-SMA were enhanced by puerarin. We also found that puerarin increased HO-1, SOD and GSH levels. CONCLUSIONS: These findings demonstrated that RPEs blocked PQ-induced Fstl 1 pathways and oxidative stress by inhibiting miR-21 expression, leading to attenuation of PQ-induced lung fibrosis.

Effects of pre-exercise feeding on serum hormone concentrations and biomarkers of myostatin and ubiquitin proteasome pathway activity.

PURPOSE: The aim of the study was to examine the acute effects of pre-exercise ingestion of protein, carbohydrate, and a non-caloric placebo on serum concentrations of insulin and cortisol, and the intramuscular gene expression of myostatin- and ubiquitin proteasome pathway (UPP)-related genes following a bout of resistance exercise. METHODS: Ten untrained college-aged men participated in three resistance exercise sessions (3 × 10 at 80 % 1RM for bilateral hack squat, leg press, and leg extension) in a cross-over fashion, which were randomly preceded by protein, carbohydrate, or placebo ingestion 30 min prior to training. Pre-supplement/pre-exercise, 2 h and 6 h post-exercise muscle biopsies were obtained during each session and analyzed for mRNA fold changes in myostatin (MSTN), activin IIB, follistatin-like 3 (FSTL3), SMAD specific E3 ubiquitin protein ligase 1 (SMURF1), forkhead box O3, F-box protein 32 (FBXO32), and Muscle RING-finger protein-1, with beta-actin serving as the housekeeping gene. Gene expression of all genes was analyzed using real-time PCR. RESULTS: Acute feeding appeared to have no significant effect on myostatin or UPP biomarkers. However, resistance exercise resulted in a significant downregulation of MSTN and FBXO32 mRNA expression and a significant upregulation in FSTL3 and SMURF1 mRNA expression (p < 0.05). CONCLUSIONS: An acute bout of resistance exercise results in acute post-exercise alterations in intramuscular mRNA expression of myostatin and UPP markers suggestive of skeletal muscle growth. However, carbohydrate and protein feeding surrounding resistance exercise appear to have little influence on the acute expression of these markers.

Resolution of inflammation induces osteoblast function and regulates the Wnt signaling pathway.

OBJECTIVE: Inflammation in the bone microenvironment stimulates osteoclast differentiation, resulting in uncoupling of resorption and formation. Mechanisms contributing to the inhibition of osteoblast function in inflammatory diseases, however, have not been elucidated. Rheumatoid arthritis (RA) is a prototype of an inflammatory arthritis that results in focal loss of articular bone. The paucity of bone repair in inflammatory diseases such as RA raises compelling questions regarding the impact of inflammation on bone formation. The aim of this study was to establish the mechanisms by which inflammation regulates osteoblast activity. METHODS: We characterized an innovative variant of a murine model of arthritis in which inflammation is induced in C57BL/6J mice by transfer of arthritogenic K/BxN serum and allowed to resolve. RESULTS: In the setting of resolving inflammation, bone resorption ceased and appositional osteoblast-mediated bone formation was induced, resulting in repair of eroded bone. Resolution of inflammation was accompanied by striking changes in the expression of regulators of the Wnt/ß-catenin pathway, which is critical for osteoblast differentiation and function. Down-regulation of the Wnt antagonists secreted frizzled-related protein 1 (sFRP1) and sFRP2 during the resolution phase paralleled induction of the anabolic and pro-matrix mineralization factors Wnt10b and DKK2, demonstrating the role of inflammation in regulating Wnt signaling. CONCLUSION: Repair of articular bone erosion occurs in the setting of resolving inflammation, accompanied by alterations in the Wnt signaling pathway. These data imply that in inflammatory diseases that result in persistent articular bone loss, strict control of inflammation may not be achieved and may be essential for the generation of an anabolic microenvironment that supports bone formation and repair.

Effects of an alleged myostatin-binding supplement and heavy resistance training on serum myostatin, muscle strength and mass, and body composition.

This study examined 12 wk of resistance training and cystoseira canariensis supplementation on serum levels of myostatin and follistatin-like related gene (FLRG) and muscle strength and body composition. Twenty-two untrained males were randomly assigned to a placebo (PLC) or myostatin binder (MYO) group in a double-blind fashion. Blood was obtained before and after 6 and 12 wk of training. PLC and MYO trained thrice weekly using 3 sets of 6 to 8 repetitions at 85 % to 90 % 1 repetition maximum. MYO ingested 1200 mg/d of cystoseira canariensis. Data were analyzed with 2-way ANOVA. After training, total body mass, fat-free mass, muscle strength, thigh volume/mass, and serum myostatin and FLRG increased for both groups (P < 0.05); however, there were no differences between groups (P > 0.05). Twelve wk of heavy resistance training and 1200 mg/d of cystoseira canariensis supplementation appears ineffective at inhibiting serum myostatin and increasing muscle strength and mass or decreasing fat mass.