Association Of Mitogen-activated Protein Kinase 1(MAPK1) And Mitogen-activated Protein Kinase Kinase 4(MAP2K4) Gene Polymorphism With Risk Of Ischemic Stroke And Is-related Traditional Chinese Medicine Syndrome

Objective: Mitogen-activated protein kinase(MAPK)Signaling Pathway plays a significant role in ischemic stroke(IS),This study was aimed to evaluate the association of the MAPK1 gene polymorphisms rs6928,rs9340,and MAP2K4 gene polymorphisms rs3826392 with risk of ischemic stroke(IS),its different TCM syndromes and the IS-related quantitative traits.Methods: This study included 816 ischemic stroke patients(465 males and 351 females)and 816 controls matched with sex-and age-(433males and 383 females).The SNPs genotyping were conducted by Sequenom Mass Array technology.Biochemical parameters of total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)were measured by standard laboratory procedures using Hitachi 7600 Automatic Biochemistry Analyzer.The SPSS version 16.0.software(SPSS Inc.,Chicago,IL,USA)was used to perform statistical analyses of the general data between case and control.PLINK software(http://pngu.mgh.harvard.edu/~purcell/plink/)was used to evaluate the association between rs6928,rs9340,and rs3826392 polymorphisms and risk of ischemic stroke and its related phenotypes.The m RNA expression of MAPK1 and MAP2K4 gene in peripheral blood mononuclear cells was detected using reverse transcriptionpolymerase chain reaction.Results: 1.Hardy-Weinberg Equilibrium(HWE)test: the genotype frequencies of the MAPK1 gene polymorphisms rs6928,rs9340,and MAP2K4 gene polymorphisms rs3826392 in the control group do not deviate from HWE(all P>0.050).2.Association analysis:(1)rs6928,rs9340 and rs3826392 polymorphisms were not statistically significantly associated with risk of IS(all P>0.050);(2)After gender-specific stratification analysis,rs6928,rs9340 and rs3826392 polymorphisms were not statistically significantly associated with risk of IS(all P>0.050).3.Association between MAPK1 and MAP2K4 gene polymorphism and TCM syndrome:(1)Wind-phlegm obstruction syndrome: no significant differences were observed in the association between rs6928,rs9340 and rs3826392 polymorphisms and risk of IS wind-phlegm obstruction syndrome(all P>0.050);After gender-specific stratification analysis,MAPK1 gene polymorphisms rs9340 was significantly associated with risk of IS wind-phlegm obstruction syndrome patients in males(recessive: P=0.047,Padj=0.043)(2)Qi-deficiency and blood-stasis syndrome: No significant differences were observed in the association between rs6928,rs9340 and rs3826392 polymorphisms and risk of IS Qi-deficiency and blood-stasis syndrome as well as after gender-specific stratification analysis(all P>0.050).4.IS-related quantitative traits analysis:(1)rs6928,rs9340 and rs3826392 polymorphisms were not significantly associated with the serum levels of TC,TG,HDL-C?LDL-C of the IS patients(all P>0.050);After gender-specific stratification analysis,rs6928 polymorphisms was significantly associated with TG level of IS patients in males(additive: Padj = 0.0.35).5.Association between MAPK1 and MAP2K4 gene polymorphism and IS-related quantitative traits with different TCM syndrome:(1)Wind-phlegm obstruction syndrome: no significant differences were observed in the association between rs6928,rs9340 and rs3826392 polymorphisms and the serum levels of TC,TG,HDL-C?LDL-C of IS wind-phlegm obstruction syndrome patients(all P>0.050);(2)Qi-deficiency and blood-stasis syndrome: MAPK1 gene polymorphisms rs6928 and rs9340 was significantly associated with TG level of IS patients(additivers6928: Padj = 0.048;dominantrs6928: Padj = 0.008;dominantrs9340: Padj = 0.041);No significant differences were observed in the association between MAP2K4 gene polymorphisms rs3826392 and the serum levels of IS Qi-deficiency and blood-stasis syndrome patients;After gender-specific stratification analysis,MAPK1 gene polymorphisms rs6928 was significantly associated with TG level of IS patients in males(additive : Padj=0.010;dominant: Padj=0.004);MAPK1 gene polymorphisms rs9340 was significantly associated with HDL-C level of IS patients in females(dominant: Padj = 0.046).6.Association between MAPK1 and MAP2K4 gene expression levels and IS: Compared with normal control group,the expression of MAPK1 gene in IS group were significantly declined(P<0.050);There wa s no statistically significant difference between the expression of MAP2K4 gene in IS group and normal control group(P>0.050).7.Association between MAPK1 and MAP2K4 gene expression levels and IS with Different TCM syndrome: There was no statistically significant difference between the expression of MAPK1 and MAP2K4 gene in IS group with Wind-phlegm obstruction syndrome and normal control group,as well as IS group with Qi-deficiency and blood-stasis syndrome and normal control group(P>0.050).8.Tthere was a statistically significant difference between the expression of MAPK1 and MAP2K4 gene in IS group with Wind-phlegm obstruction syndrome and with Qi-deficiency and blood-stasis syndrome(P<0.050);The expression level of m RNA MAPK1 was significantly lower in patients with wind phlegm and blood stasis syndrome carrying rs6928 CC genotype than other genotype carriers(P = 0.006).Conclusions: MAPK1 gene polymorphisms rs6928,rs9340,and MAP2K4 gene polymorphisms rs3826392 may be not associated with the susceptibility of IS in Chinese Han population from Guangxi area.However,MAPK1 gene rs9340 polymorphism may affect the occurrence of ischemic stroke with wind phlegm and blood stasis syndrome in Chinese Han male in Guangxi,China.MAPK1 gene polymorphism affects the blood lipid metabolism in ischemic stroke with Qi deficiency and blood stasis.