Effects Of Tetramethylpyrazine On Calmodulin-dependent Protein Kinase ? Delta Isoform Promote Rats Vascular Remodeling

Objective:To observe the effects of tetramethylpyrazine(TMP)on Calmodulin dependent protein kinase ? delta isoform promote rats vascular remodeling.TMP is the effective component of Ligusticum Chuanxiong Hort.We use TMP intervene primary cultured rat aortic smooth muscle cells and observation the effects of TMP on the primary cultured rat aortic smooth muscle cells proliferation,migration and apoptosis.To explore the relationship of CaMKII-delta isoform and miRNAs(miRNA-21?miRNA-221?miRNA-223?miRNA-143 and miRNA-145)and the mechanism of TMP in inhabit vascular remodeling.Methods:SD rat's thoracic sorta VSMCs in vitro cultured were divided into four groups.The control group?the low dose TMP(50umol/L)group?the middle dose TMP(100umol/L)group?the high dose TMP(200umol/L)groug.VSMCs was incubated different concentrations of TMP for 48 hours.The proliferation ratio was detected by cck-8.The migration was measured by transwell.Flow cytometry instrument is used to test the change of cells apoptosis.Gene and protein expressions of miRNAs(miRNA-21?miRNA-221?miRNA-223?miRNA-143 and miRNA-145)and CaMK???CaMKIIN were detected with real-time fluorescent quantitative PCR and Westernblot respectively.Results:Compared with the control group.the proliferation and migration ratio of VSMCs obviously declined in the different concentrations of TMP(P<0.05).The ratio of apoptosis VSMCs was obviously increased in the different concentrations of TMP(P<0.05).Comparedwith the control group.the gene expressions of miRNA-21?miRNA-221?miRNA-223 and CaMK?? were obviously down-regulation(P<0.05).the gene expressions of miRNA-143,miRNA-145 and CaMKIIN were obviously up-regulation(P<0.05).Protein expression of CaMK?? was obviously down-regulation(P<0.05)and CaMKIIN were obviously up-regulation(P<0.05).The result has the statistically significant(P<0.05).The aforesaid indices were dose-dependent in the low?middle and high dose TMP.Conclusion:TMP can inhabit the proliferation?migration and promote apoptosis by down-regulation the gene expressions of miRNA-21?miRNA-221?miRNA-223 and up-regulation the gene expressions of miRNA-143?miRNA-145 and then inhabit the expression of CaMK?? and up-regulation the expression of CaMKIIN.This maybe the mechanism of TMP inhabit vascular remodeling.We can infer CaMK?? is one of the important target genes of specific miRNAs(miRNA-21?miRNA-221?miRNA-223?miRNA-143?miRNA-145).