Association Study Of CAV1 Rs1049337 Polymorphism And Phlegm-blood Stasis Syndrome Of Ischemic Stroke And Coronary Heart Disease

Objective:Previous studies have shown that CAV1 gene plays an important role in the pathological process of ischemic stroke（IS）and coronary heart disease（CHD）.Phlegm-blood stasis syndrome is an important TCM syndrome type in IS and CHD.However,the effect of CAV1 3’UTR polymorphisms on risk of IS and CHD remains unknow.Thus,this current study aims to explore the association among CAV1 gene expression and rs1049337 polymorphism and phlegm-blood stasis syndrome of IS and CHD,as well as clinical indicators.Methods:This case-control study contained 550 IS patients with phlegm-blood stasis syndrome,550 CHD patients with phlegm-blood stasis syndrome and 550 controls.Genotyping determination for rs1049337 of CAV1 was performed by Sequenom MassARRAY iPLEX.Dual-luciferase reporter assay was conducted to confirm the binding of studied miRNA and CAV1.Expression level of CAV1 mRNA was detected by quantitative reverse transcription polymerase chain reaction（qRT-PCR）.Clinical biochemical indicators were detected by HITACHI 7600 automatic biochemical analyzer,Mindray 6900 automatic blood cell analyzer and StAgO Capact blood coagulation instrument.Statistical analysis was done by Plink and SPSS sofrware.Results:1、Genotypic frequencies and Hardy-Weinberg Equilibrium（HWE）TestResults of HWE test indicated that,the frequencies of CAV1rs1049337 genotypes distribution were in HWE in control group,as well as male and female control subgroups.The frequencies of rs1049337 CC,CT,TT genotypes were significantly different between CHD patients with phlegm-blood stasis syndrome and controls（χ2=8.710,P=0.013）.2、Association analyses of CAV1 rs1049337 and susceptibility of phlegm-blood stasis syndrome with IS and CHDIn dominant model,rs1049337 was significantly associated with phlegm-blood stasis syndrome of IS risk(OR=0.75,95%CI=0.58-0.97,P=0.030;after adjusting for sex and age:OR_adj=0.76,95%CI=0.59-0.98,P_adj=0.032).And significant associations were found between rs1049337and phlegm-blood stasis syndrome of CHD risk in additive(OR=0.77,95%CI=0.64-0.92,P=0.003;after adjusting for sex and age:OR_adj=0.77,95%CI=0.64-0.92,Padj=0.004),dominant(OR=0.71,95%CI=0.55-0.92,P=0.009;after adjusting for sex and age:OR_adj=0.71,95%CI=0.55-0.92,P_adj=0.009),recessive(OR=0.71,95%CI=0.51-0.98,P=0.040;after adjusting for sex and age:OR_adj=0.71,95%CI=0.51-0.99,P_adj=0.043),and allelic models（OR=0.78,95%CI=0.09-0.66,P=0.004）.3、Correlation analysis between CAV1 rs1049337 and related clinical indicators in IS patients with phlegm-blood stasis syndromeCAV1 rs1049337 was positively correlated to serum UA level in IS patients with phlegm-blood stasis syndrome（P=0.025）.After gender stratification analysis,CAV1 rs1049337 was associated with diastolic blood pressure（DBP）,very low density lipoprotein（VLDL）,triglyceride（TG）in male IS patients with phlegm-blood stasis syndrome（all P<0.050）,and triglyceride cholesterol（TC）,thrombin time（TT）in female IS patients with phlegm-blood stasis syndrome（both P<0.050）.4、Correlation analysis between CAV1 rs1049337 and related clinical indicators in CHD patients with phlegm-blood stasis syndromeSignificant correlation was found between rs1049337 and serum CRP level in CHD patients with phlegm-blood stasis syndrome（P=0.014）.After gender stratification analysis,CAV1 rs1049337 was associated with P2hPG in male CHD patients with phlegm-blood stasis syndrome(P_adj=0.006).5、Comparison of CAV1 mRNA expression level in phlegm-blood stasis syndrome with IS and CHD and controlCHD patients exhibited a lower CAV1 expression level than controls（P<0.001）.6、Results of dual-luciferase report assayResults of dual-luciferase report assay confirmed the binding of miR-1285 and CAV1 at rs1049337.Conclusions:CAV1 rs1049337 might be a shared risk loci in phlegm-blood stasis syndrome of both IS and CHD.The underlying pathological mechanism might be due to the mutation of rs1049337 affecting the binding efficiency of miR-1285 and CAV1 gene,thus affecting the susceptibility of phlegm-blood stasis syndrome of both IS and CHD.Besides,CAV1 rs1049337 might participate in the occurrence of IS with phlegm-blood stasis syndrome through affecting UA levels,blood pressure fluctuation and lipid metabolism in male,lipid metabolism and coagulation in female.CAV1 gene rs1049337 polymorphism might affect the occurrence of CHD with phlegm-blood stasis syndrome by regulate C-reactive protein levels,and blood glucose fluctuation in male.