Effect And Mechanism Of Bisdemethoxycurcumin On Mast Cell Mediated Allergic Diseases

Allergic disease is a pathological immune response to tissue dysfunction caused by antigen stimulation.It eventually leads to inflammation and tissue damage.It HSA rapid onset and strong reaction.Mast cells play an important role in the development of allergic diseases.When antigens repeatedly attack the body,the antigens are presented to B cells after being processed by the antigen-presenting cells,prompting B cells to secrete antigen-specific Ig E antibodies,antigen-specific Ig E.The antibody binds to receptor FCεRI on the surface of mast cells,resulting in an allergic reaction.Objections: To explore the effect of disdemethoxycurcumin on DNP-Ig E/HSA-induced mast cell degradulations model,OVA induced mouse allergic rhinitis model,Compound 48/80 induced mouse anaphylactic shock model,OVA induced active systemic immune disease model.Methods: In vitro experiments: 1.Establishment of DNP-Ig E/HSA induced degranulation model of RBL-2H3 cells The effects of different cell densities,DNP-Ig E concentration,stimulation time,and concentration of DNP-HSA on the degranulation model of RBL-2H3 cells were investigated.2.To investigate the effect of curcuminoids on the cell viability of the RBL-2H3 degranulation model.Using the optimal stimulation conditions determined in the above steps,the effects of various doses of curcuminoids on cell proliferation in DNP-Ig E/HSA-induced RBL-2H3 cell degranulation model were determined.(1)Using the optimal stimulation conditions determined in the above steps,the effects of various dose groups of curcuminoids on the release of β-HEX by DNP-Ig E/HSA-induced degranulation model of RBL-2H3 cells were investigated.(2)Toluidine blue staining was used to observe the effect of the above test substance on the degree of cell degranulation.(3)F-actin microfilament staining was used to explore the effect of BDMC on the rearrangements of F-actin cytoskeleton.In vivo experiments: 1.To investigate the effect of disdemethoxycurcumin on OVA-induced allergic rhinitis model in mice(1)Observe the number of nasal rubbing of allergic rhinitis mice after daily challenge.(2)Collect serum to determine OVA-specific Ig E levels.(3)The nasal mucosa was pathologically sectioned,and then stained by H&E for the observation of pathological changes,and stained by toluidine blue for exploration mast cell levels and degranulation.2.To investigate the effect of disdemethoxycurcumin on Compound 48/80 induced anaphylactic shock in mice After intraperitoneal administration of Compound 48/80,the survival rate of each group of mice within 1 hour was observed.3.To investigate the effect of disdemethoxycurcumin on OVA-induced mouse systemic immune disease model After the last challenge,rectal temperature changes were determined in mice within 80 min.3.To investigate the effect of curcuminoids on the degranulation of RBL-2H3 degranulation modelResults: In vitro experiments: 1.Establishment of DNP-Ig E/HSA induced degranulation model of RBL-2H3 cells Finally,the experimental conditions were determined as follows: RBL-2H3 was plated in 48-well plates at 6×105 cell/m L,and after being adhered overnight.The supernatant was discarded and stimulated with 100ng/m L of DNP-Ig E for 12 hours before discarding the supernatant.250 ng/m L of DNP-HSA was added for 1 h and then the reaction was stopped in an ice bath for 10 min.After centrifugation at 300 ×g for 10 min,50 μL supernatant was added in 50 μL of the substrate for 1.5 h.The 405 nm OD was measured.2.To investigate the effect of curcuminoids on the cell viability of the RBL-2H3 degranulation model.Curcumin compounds in each dose group(6.3μM-73μM)did not reduce the cell viability of DNP-Ig E/HSA induced RBL-2H3 cell degranulation model.3.To investigate the effect of curcuminoids on the degranulation of RBL-2H3 degranulation model(1)To investigate the effect of curcuminoids on β-Hex levels Curcumin,demethoxycurcumin and disdemethoxycurcumin all decreased the release rate of β-HEX to some extent.The IC50 were curcumin 99.47 μM,demethoxycurcumin 30.15 μM,and bisdesmethoxycurcumin 19.27 μM,respectively.(2)To investigate the effect of curcuminoids on cell degranulation Bisdemethoxycurcumin inhibit the shape changes and reduce the release of purple granule of RBL-2H3 cells induced by DNP-Ig E/HSA.(3)To explore the effect of BDMC on the rearrangements of F-actin cytoskeleton The results suggest that Bisdemethoxycurcumin suppress rearrangements of F-actin cytoskeleton.In vivo experiments: 1.To investigate the effect of disdemethoxycurcumin on OVA-induced Allergic Rhinitis Model in Mice(1)Bisdemethoxycurcumin significantly inhibited the number of nasal rubbing of mice with OVA-induced allergic rhinitis(P<0.05).(2)Bisdemethoxycurcumin significantly reduce OVA-specific Ig E levels in mice with OVA-induced allergic rhinitis(P<0.05).(3)Bisdemethoxycurcumin markedly ameliorate nasal mucosa edema,exuviation,goblet cell hyperplasia,inflammatory cellular infiltration,and mast cell degranulation.2.To investigate the effect of disdemethoxycurcumin on Compound 48/80 induced anaphylactic shock in mice Bisdemethoxycurcumin significantly increased the survival rate of Compound 48/80-induced anaphylactic shock model mice(P<0.05).3.To investigate the effect of disdemethoxycurcumin on OVA-induced mouse systemic immune disease model Bisdemethoxycurcumin significantly inhibited the reduction of rectal temperature in OVA-induced active systemic immune disease mice(P<0.05).Conclusion: 1.Among the curcuminoids,bisdemethoxycurcumin HSA the greatest inhibitory effect on Ig E-induced mast cell activation,which can significantly reduce the degree of degranulation of mast cells.2.Bisdemethoxycurcumin inhibits mast cell-mediated allergic diseases,including OVA-induced allergic rhinitis in mice,Compound 48/80-induced anaphylactic shock in mice,and inhibition of OVA-induced active systemic immune disease in mice.