Abstract
Background: Melittin, the major medicinal component of honeybee venom, exerts antiinflammatory, analgesic, and anti-arthritic effects in patients with Rheumatoid Arthritis (RA). RA is an inflammatory autoimmune joint disease that leads to irreversible joint destruction and functional loss. Fibroblast-Like Synoviocytes (FLS) are dominant, special mesenchymal cells characterized by the structure of the synovial intima, playing a crucial role in both the initiation and progression of RA.
Objective: In this study, we evaluated the effects of melittin on the viability and apoptosis of FLS isolated from patients with RA.
Methods: Cell viability was determined using CCK-8 assays; apoptosis was evaluated by flow cytometry, and the expression levels of apoptosis-related proteins (caspase-3, caspase-9, BAX, and Bcl-2) were also determined. To explore whether melittin alters inflammatory processes in RA-FLS, IL-1β levels were determined using an enzyme-linked immunosorbent assay (ELISA). Furthermore, we performed GFP-LC3 punctate fluorescence dot assays and western blotting (for LC3, ATG5, p62, and Beclin 1) to assess autophagy in RA-FLS.
Results: Our results show that melittin can significantly impair viability, promote apoptosis and autophagy, and inhibit IL-1β secretion in RA-FLS.
Conclusion: Melittin may be useful in preventing damage to the joints during accidental local stimulation.
Keywords: Melittin, rheumatoid arthritis, fibroblast-like synoviocytes, autophagy, apoptosis, ELISA.
Graphical Abstract
[PMID: 23327517]
[http://dx.doi.org/10.2146/ajhp050514] [PMID: 17158693]
[http://dx.doi.org/10.1038/nri846] [PMID: 12094227]
[http://dx.doi.org/10.1111/j.0105-2896.2009.00859.x] [PMID: 20193003]
[http://dx.doi.org/10.1186/ar1153] [PMID: 15059280]
[http://dx.doi.org/10.1111/j.0105-2896.2009.00854.x] [PMID: 20193004]
[http://dx.doi.org/10.1007/s10495-009-0317-y] [PMID: 19199037]
[http://dx.doi.org/10.1006/cyto.1999.0552] [PMID: 10704256]
[http://dx.doi.org/10.1101/gad.1599207] [PMID: 18006683]
[http://dx.doi.org/10.1074/jbc.M113.462408] [PMID: 23576438]
[http://dx.doi.org/10.3389/fimmu.2018.01577] [PMID: 30072986]
[http://dx.doi.org/10.1002/art.38190] [PMID: 24449574]
[http://dx.doi.org/10.1007/s00726-014-1833-9] [PMID: 25200682]
[http://dx.doi.org/10.3390/molecules21050616] [PMID: 27187328]
[http://dx.doi.org/10.1016/j.jtcms.2016.09.005]
[http://dx.doi.org/10.1080/01926230701320337] [PMID: 17562483]
[http://dx.doi.org/10.1038/cddis.2016.490] [PMID: 28102843]
[PMID: 29652658]
[http://dx.doi.org/10.1007/978-3-642-79437-7_8]
[http://dx.doi.org/10.1161/01.HYP.32.2.280] [PMID: 9719055]
[http://dx.doi.org/10.1038/sj.cdd.4400476] [PMID: 10200555]
[http://dx.doi.org/10.1006/cyto.1998.0372] [PMID: 9770330]
[http://dx.doi.org/10.1016/j.molimm.2007.02.001] [PMID: 17353049]
[http://dx.doi.org/10.1042/bse0550105] [PMID: 24070475]
[http://dx.doi.org/10.1038/ncb1007-1102] [PMID: 17909521]
[http://dx.doi.org/10.3389/fimmu.2017.00165] [PMID: 28289410]
[http://dx.doi.org/10.1242/jcs.02735] [PMID: 16390869]
[http://dx.doi.org/10.1007/978-1-59745-157-4_4]
[http://dx.doi.org/10.4161/auto.1.2.1858] [PMID: 16874032]
[http://dx.doi.org/10.1016/j.ymeth.2014.11.021] [PMID: 25484342]