The Mechanism Of Trimethylamine-N-oxide Regulating The Inflammatory Response Of Smooth Muscle Cells Through SM22α

Objective: To investigate whether trimethylamine-n-oxide(TMAO)can promote the inflammatory response of vascular smooth muscle cells(VSMCs)by inhibiting smooth muscle 22 alpha(SM22α),and to provide a basis for future clinical treatment.Methods:1.VSMCs were treated with 0 μmol/L and 50 μmol/L TMAO respectively to verify the effect of TMAO on VSMCs.VSMCs were treated with the optimal concentration of TMAO for 0,6,12 and 24 h,and used as the time gradient group.SM22α was detected by Western blot,and the optimal time of TMAO on VSMCs were selected.2.VSMCs were treated with 100 μmol/L TMAO for 24 h,and the expressions of IL-1β,IL-6 and TNF-α were detected by Western blot to determine the effect of TMAO on inflammatory cytokines of VSMCs.3.VSMCs were transfected with p EX3-SM22α for 24 h to overexpress SM22α,and then treated with TMAO.The expression of the above-mentioned inflammatory factors was detected by Western blot to determine whether TMAO regulates the expression of inflammatory factors through SM22α.Results: 1.TMAO inhibited the expression of SM22α in VSMCs.Compared with the Con group,TMAO significantly inhibited the expression of SM22α in VSMCs,and the difference was statistically significant(P<0.005).2.Western blot results showed that TMAO could promote the expression of IL-1β in VSMCs compared with Con group,and the difference was statistically significant(P<0.05).The expression of IL-6 and TNF-α was significantly increased,and the difference was statistically significant(P<0.005).3.VSMCs were transfected with SM22α overexpressed plasmid vector,and the expression of SM22α was detected by Western blot.The results showed that the expression of SM22α in VSMCs in p EX3-SM22α+TMAO group was higher than that in p EX3-Con +TMAO group,and the difference was statistically significant(P<0.005).4.After TMAO treatment,the expression levels of IL-1β,IL-6 and TNF-α in VSMCs overexpressing SM22α were decreased in Pex3-SM22α +TMAO group compared with Pex3-Con +TMAO group,with statistical significance(P<0.05).Conclusions: TMAO can promote the inflammatory response and it promotes the inflammatory response by inhibiting SM22α.