In healthy joints, synovial fibroblasts (SFs) provide the microenvironment required to mediate homeostasis, but these cells adopt a pathological function in rheumatoid arthritis (RA). Carbohydrates (glycans) on cell surfaces are fundamental regulators of the interactions between stromal and immune cells, but little is known about the role of the SF glycome in joint inflammation. Here we study stromal guided pathophysiology by mapping SFs glycosylation pathways. Combining transcriptomic and glycomic analysis, we show that transformation of fibroblasts into pro-inflammatory cells is associated with glycan remodeling, a process that involves TNF-dependent inhibition of the glycosyltransferase ST6Gal1 and α2-6 sialylation. SF sialylation correlates with distinct functional subsets in murine experimental arthritis and remission stages in human RA. We propose that pro-inflammatory cytokines remodel the SF-glycome, converting the synovium into an under-sialylated and highly pro-inflammatory microenvironment. These results highlight the importance of glycosylation in stromal immunology and joint inflammation.
24 January 2022 in Molecular Nutrition & Food Research
Brazilian green propolis is a complex mixture of natural compounds that is difficult to analyze and standardize; as a result, controlling its quality is challenging. In this study, we used the positive and negative modes of ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time of flight mass spectrometry in conjunction with high-performance liquid chromatography for the identification and characterization of seven phenolic acid compounds in Brazilian green propolis. The optimal operating conditions for the electrospray ionization source were capillary voltage of 3500 V and drying and sheath gas temperatures of 320 °C and 350 °C, respectively. Drying and sheath gas flows were set to 8 L/min and 11 L/min, respectively. Brazilian green propolis was separated using the HPLC method, with chromatograms for samples and standards measured at 310 nm. UPLC-ESI-QTOF-MS was used to identify the following phenolic compounds: Chlorogenic acid, caffeic acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, caffeic acid phenethyl ester (CAPE), and artepillin C. Using a methodologically validated HPLC method, the seven identified phenolic acids were then quantified among different Brazilian green propolis. Results indicated that there were no significant differences in the content of a given phenolic acid across different Brazilian green propolis samples, owing to the same plant resin sources for each sample. Isochlorogenic acid B had the lowest content (0.08 ± 0.04) across all tested Brazilian green propolis samples, while the artepillin C levels were the highest (2.48 ± 0.94). The total phenolic acid content across Brazilian green propolis samples ranged from 2.14–9.32%. Notably, artepillin C quantification is an important factor in determining the quality index of Brazilian green propolis; importantly, it has potential as a chemical marker for the development of better quality control methods for Brazilian green propolis.
A comparative study between Chinese propolis and Brazilian green propolis showed that they have a distinct chemome but similar antioxidant and anti-inflammatory potential.
Pectolinarin and linarin are two major flavone O-glycosides of Cirsium japonicum, which has been used for thousands of years in traditional Chinese medicine. Pharmacological research on pectolinarin and linarin is meaningful and necessary. Here, a process for the purification of pectolinarin and linarin from C. japonicum was established using macroporous resin enrichment followed by prep-HPLC separation. The results show the purity of pectolinarin and linarin reached 97.39% and 96.65%, respectively. The in vitro bioactivities result shows the ORAC values of pectolinarin and linarin are 4543 and 1441 µmol TE/g, respectively, meanwhile their inhibition rate of BSA-MGO-derived AGEs is 63.58% and 19.31% at 2 mg/mL, which is 56.03% and 30.73% in the BSA-fructose system, respectively. The COX-2 inhibition rate at 50 µg/mL of linarin and pectolinarin reached 55.35% and 40.40%, respectively. Furthermore, the in vivo bioassay combining of histopathologic evaluation and biochemical analysis of liver glutamic oxaloacetic transaminase, serum creatinine and TNF-α show pectolinarin can alleviate lipopolysaccharide (LPS)-induced acute liver and kidney injury in mice. Metabolomics analysis shows that pectolinarin attenuates LPS-challenged liver and kidney stress through regulating the arachidonic acid metabolism and glutathione synthesis pathways. Collectively, our work presents a solid process for pectolinarin and linarin purification and has discovered a promising natural therapeutic agent—pectolinarin.