ABSTRACT
Objective The changes of Mg2+ concentration and cell apoptosis were detected by using siRNA to silence MagT1 in rat cardio myocytes.Methods MagT1 siRNA sequences were designed and synthetized,then transfected into primary cultured rat myocardial cell for silencing MagT1.The expression of MagT1 mRNA and protein were detected by RT-PCR and Western blot.The changes of Mg2+ concentration in the cells were detected by fluorescence microscopy.Flow cytometry (FCM) was used to detect the cell apoptosis.Results Compared with negative siRNA group,MagT1 siRNA transfected rat cardiomyocytes after 48 h,MagT1 mRNA silence efficiency was 51.83 ％ (P＜0.05),the silence of MagT1 protein efficiency was 56.75 ％ (P＜0.05),intracellular Mg2+ concentration was reduced by 29.13％ (P＜0.05),the apoptosis rate was 31.18％ (P＜0.01);MagT1 siRNA transfected rat cardiomyocytes after 60 h,MagT1 mRNA silence efficiency was 86.91％ (P＜0.01),the silence of MagT1 protein efficiency was 83.85％ (P＜0.01),intracellular Mg2+ concentration was reduced by 41.32％ (P＜0.01),the apoptosis rate was 40.61％ (P＜0.01).Conclusion After the silencing of MagT1,the concentration of Mg2+ in the cells decreased significantly,the apoptotic rate increased significantly,cell life activities are greatly affected.
ABSTRACT
Objective To investigate the correlation of ERα gene PvuⅡ ,XbaⅠ and ERβ gene RsaⅠ ,AluⅠ digestion polymorphism with coronary atherosclerotic heart disease(CHD) risk factors in Yancheng area .Methods A total of 124 cases of CHD and 163 persons undergoing physical examination served as the CHD group and CON group .The enzyme method was adopted to detect TG and TC .The direction method was adopted to detect HDL and LDL .ERα gene PvuⅡ ,XbaⅠ and ERβ gene RsaⅠ ,AluⅠ digestion polymorphisms were detected by adopting RFLP-PCR .Results The ratios of smoking history ,family history ,complicating hypertension and diabetes ,and the level of body mass index ,TC ,TG and LDLC in the CHD group were significantly higher than those in the control group ,the difference was statistically significant (P＜0 .05) .The various indicators had no statistically difference between male and female(P＞0 .05) .The frequency distribution and geographic distribution of ERα gene PvuⅡ ,XbaⅠ and ERβ gene RsaⅠ ,Alu Ⅰ digestion polymorphisms had no difference between the two groups ,all conformed to Hardy-Weinberg genetic equilibrium and had the group representativeness .pp ,xx ,RR and AA genotypes in the CHD group were maximal ,while PP , XX ,rr and aa genotypes were minimal ;Pp ,xx ,RR and AA genotypes in the CON group were maximal ,while PP ,XX ,rr and aa genotypes were minimal .The distribution frequency of p and x genes in the CHD group was significantly higher than that in the control group ,the difference was statistically significant(P＜0 .05) .Conclusion The estrogen gene polymorphism might be a target spot for effectively treating CHD ,and p and x gene distribution frequency may be related with CHD risk factors .