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Abstract Background Pharmacological modulation of cannabinoid 2 receptor (CB2R) is a promising therapeutic strategy for pulmonary fibrosis (PF). Thus, to develop CB2R selective ligands with new chemical space has attracted much research interests. This work aims to discover a novel CB2R agonist from an in-house library, and to evaluate its therapeutic effects on PF model, as well as to disclose the pharmacological mechanism. Methods Virtual screening was used to identify the candidate ligand for CB2R from a newly established in-house library. Both in vivo experiments on PF rat model and in vitro experiments on cells were performed to investigate the therapeutic effects of the lead compound and underlying mechanism. Results A “natural product-like” pyrano[2,3-b]pyridine derivative, YX-2102 was identified that bound to CB2R with high affinity. Intraperitoneal YX-2102 injections significantly ameliorated lung injury, inflammation and fibrosis in a rat model of PF induced by bleomycin (BLM). On one hand, YX-2102 inhibited inflammatory response at least partially through modulating macrophages polarization thereby exerting protective effects. Whereas, on the other hand, YX-2102 significantly upregulated CB2R expression in alveolar epithelial cells in vivo. Its pretreatment inhibited lung alveolar epithelial-to-mesenchymal transition (EMT) in vitro and PF model induced by transforming growth factor beta-1 (TGF-β1) via a CB2 receptor-dependent pathway. Further studies suggested that the Nrf2-Smad7 pathway might be involved in. Conclusion These findings suggest that CB2R is a potential target for PF treatment and YX-2102 is a promising CB2R agonist with new chemical space.

Summary: The current procedure thoroughly explains how to reprogram induced pluripotent stem cells (iPSCs) from the patient’s peripheral blood mononuclear cells (PBMCs), a less invasive source; e.g., somatic cells. We describe how to isolate PBMCs and reprogram them into iPSCs by electroporation. Furthermore, we provide an alternative approach to generating iPSC using Geltrex or Matrigel matrix to replace MEF-feeder. The challenge of this process is the relatively lower cell survival rates of PBMCs due to the damage of electroporation.For complete details on the use and execution of this protocol, please refer to Hu et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Abstract Radiation-induced lung fibrosis (RILF) is a long-term adverse effect of curative radiotherapy. The accumulation of myofibroblasts in fibroblastic foci is a pivotal feature of RILF. In the study, we found the inhibitory effect of grape seed proanthocyanidins (GSPs) on irradiation-induced differentiation of human fetal lung fibroblasts (HFL1). To explore the mechanism by which GSPs inhibit fibroblast differentiation, we measured the reactive oxygen species (ROS) levels, mitochondrial function, mitochondrial dynamics, glycolysis and the signaling molecules involved in fibroblast transdifferentiation. GSPs significantly reduced the production of cellular and mitochondrial ROS after radiation. The increases in mitochondrial respiration, proton leak, mitochondrial ATP production, lactate release and glucose consumption that occurred in response to irradiation were ameliorated by GSPs. Furthermore, GSPs increased the activity of complex I and improved the mitochondrial dynamics, which were disturbed by irradiation. In addition, the elevation of phosphorylation of p38MAPK and Akt, and Nox4 expression induced by irradiation were attenuated by GSPs. Blocking Nox4 attenuated irradiation-mediated fibroblast differentiation. Taken together, these results indicate that GSPs have the ability to inhibit irradiation-induced fibroblast-to-myofibroblast differentiation by ameliorating mitochondrial dynamics and mitochondrial complex I activity, regulating mitochondrial ROS production, ATP production, lactate release, glucose consumption and thereby inhibiting p38MAPK-Akt-Nox4 pathway.

Caffeic acid (CA) could be considered as an important natural antioxidant. However, the low solubility and stability of CA in various solvent systems is a major limiting factor governing its overall application in the lipid industry. However, the synthesis of methyl caffeate (MC) using CA and methanol is a feasible way to improve its lipophilicity. Herein, the reaction conditions and kinetic parameters for the synthesis of MC using p-toluenesulfonic acid (PTSA) as a catalyst were investigated, and the product was confirmed by liquid chromatography--mass spectrometry (LC--MS), Fourier transform infrared (FTIR) and nuclear magnetic resonance (NMR) spectroscopy, and melting point analysis. The highest yield of MC catalyzed by PTSA attained 84.0 % under the optimum conditions as follows: molar ratio of methanol to CA of 20:1, reaction temperature of 65 °C, mass ratio of catalyst to substrate of 8 %, and a reaction time of 4 h. The esterification kinetics of CA and methanol is described by the pseudo-homogeneous second order reversible model. The relationship between temperature and the forward rate constant gives the activation energy of 17.5 kJ mol-1. These results indicated that PTSA possesses high catalytic activity in the synthesis of MC, which is an efficient catalyst suitable for MC production in the chemical industry. [ABSTRACT FROM AUTHOR]