Basic & Clinical Medicine ›› 2016, Vol. 36 ›› Issue (3): 331-336.

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Inhibition of nitric oxide on the activation of NLRP3 inflammasome myoblast /myotubes stimulated by IFN-γ in vitro

  

  • Received:2015-06-15 Revised:2015-12-02 Online:2016-03-05 Published:2016-02-22
  • Supported by:
    ; the Science and Technology Planning Project of Guangdong Province, People’s Republic of China

Abstract: Objective: To observe the effect of sodium nitroprusside (SNP) or nitro L arginine acid methyl ester (L-NAME) on the NLRP3 inflammasome activation of C2C12 myoblast or differentiated myotubes in inflammatory culture in vitro. Methods: C2C12 myoblasts were stimulated by IFN-γ, then the formation and activation of NLRP3 inflammasome were analyzed by qPCR and Western blot,and the secretion of IL-1β in the cell culture supernatant was detected by ELISA. Further, C2C12 myoblasts induced by IFN-γ were treated by L-NAME or SNP respectively, then the formation and activation of NLRP3 inflammasome and the secretion of IL-1β were analyzed. Results: qPCR and Western blotting detection confirmed that expression of NLRP3, ASC and mature-caspase-1 in C2C12 myoblasts or differentiation myotubes induced by IFN-γ was up-regulated(P<0.01). ELISA analysis further confirmed that the IL-1β concentration in the medium of C2C12 induced by IFN-γ was up-regulated compared with that of the non stimulating cells(P<0.01).At 6 h after SNP treatment, a significant down-regulation of mRNA and protein levels of differentiation myotube inflammasome (ASC and NLRP3, caspase-1) was found(P<0.01) and a contrary result was obtained after L-NAME treatment(P<0.05). Consisvent with the above results, the expression of IL-1βin the culture supernantent was down-regulated or up-regulated by SNP or L-NAME respectively(P<0.05). Conclusions: In inflammatory conditions, C2C12 myoblasts or differentiation myotubes possess the ability to synthesize inflammasome NLRP3. NO might cause a certain inhibition on the NLRP3 inflammasome formation and activation.

Key words: Nitric Oxide, C2C12 myoblast , Inflammasomes

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