孕期吸烟对SD大鼠胚胎心脏发育的影响
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摘要
目的探讨孕期吸烟对SD大鼠胚胎心脏发育的影响及分子机制。方法将24只怀孕SD母鼠随机分为4组:正常对照组、10支烟/d组、20支烟/d组、40支烟/d组,每组6只,并于孕期0.5 d(GD 0.5)后开始给予吸烟处理直至分娩。采用超声多普勒Vevo2000检测出生后1 d的新生乳鼠的左心结构及其心脏功能;采用Color-Doppler检测心室血流方向,以确认室间隔的发育是否异常。心功能检测完毕后,收集各组新生乳鼠心脏组织,采用Western-blot检测心肌特异转录因子GATA4的表达量。结果各组新生乳鼠心脏重量无明显差异,40支烟/d处理组乳鼠体重显著下降,与对照组相比,差异有统计学意义(P<0.001)。孕期持续给予40支烟/d被动吸烟处理能显著降低左心室重量、左心室舒张/收缩末期前壁厚度和左心室舒张/收缩末期后壁厚度,差异均有统计学意义(P<0.05)。孕期给予20支烟/d和40支烟/d处理后,新生乳鼠的GATA4蛋白表达呈剂量依赖性下调,40支烟/d组与对照组相比,差异有统计学意义(P<0.05)。结论母鼠在怀孕期间接受被动吸烟后,引发后代新生乳鼠的心功能受损,其机制可能与下调心肌特异转录因子GATA4表达相关。
Objective To investigate the effect of maternal smoking on cardiac development in Sprague Dawley rats and the underlying mechanism. Methods Pregnant Sprague Dawley rats(SD rats) were randomly dived into 4 groups: Control, 10cigarettes / day, 20 cigarettes / day, 40 cigarettes / day, and exposed to passive cigarettes smoking throughout gestation. On postnatal day 1(PND1), echocardiography was performed to assess the left ventricular structure and cardiac function in neonatal rats. Color Doppler was used to detect the blood direction and velocity and confirm the presence of ventricular septum.Neonatal heart tissues from each group were collected after echocardiography assessment, and western blot was used to detect the expression level of cardiac transcriptional factor GATA4. Results(1) The average of heart weight had no significant change between each group, however the body weight decreased significantly in 40 cigarettes / day group(P<0.001).(2) Maternal smoking to 40 cigarettes daily significantly reduced the weight of left ventricular(LV), LV diastolic / systolic anterior and LV diastolic / systolic posterior wall(P<0.05).(3) Maternal smoking to 40 cigarettes / day significantly down-regulated the expression of GATA4(P<0.05). Conclusion Maternal smoking induced cardiac malfunction in rat offspring, which might correlate with down regulation of GATA4 expression.
Objective To investigate the effect of maternal smoking on cardiac development in Sprague Dawley rats and the underlying mechanism. Methods Pregnant Sprague Dawley rats(SD rats) were randomly dived into 4 groups: Control, 10cigarettes / day, 20 cigarettes / day, 40 cigarettes / day, and exposed to passive cigarettes smoking throughout gestation. On postnatal day 1(PND1), echocardiography was performed to assess the left ventricular structure and cardiac function in neonatal rats. Color Doppler was used to detect the blood direction and velocity and confirm the presence of ventricular septum.Neonatal heart tissues from each group were collected after echocardiography assessment, and western blot was used to detect the expression level of cardiac transcriptional factor GATA4. Results(1) The average of heart weight had no significant change between each group, however the body weight decreased significantly in 40 cigarettes / day group(P<0.001).(2) Maternal smoking to 40 cigarettes daily significantly reduced the weight of left ventricular(LV), LV diastolic / systolic anterior and LV diastolic / systolic posterior wall(P<0.05).(3) Maternal smoking to 40 cigarettes / day significantly down-regulated the expression of GATA4(P<0.05). Conclusion Maternal smoking induced cardiac malfunction in rat offspring, which might correlate with down regulation of GATA4 expression.
引文
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[2]肖燕燕.小儿先天性心脏病诊治的进展[J].中国临床医生,2013,41(5):13-14.Xiao YY.The progress of diagnosis and treat-ment of children with congenital heart disease[J].Chinese Journal for Clinician,2013,41(5):13-14.
[3]Vander BT,Zomer AC,Zwinderman AH,et al.The changing epidemiology of congenital heart disease[J].Nat Rev Car,2011,8(1):50-60.
[4]Padula AM,Tager IB,Carmichael SL,et al.Ambient air pollution and traffic exposures and congenital heart defects in the san joaquin valley of california[J].Paed Per Epid,2013,27(4):329-339.
[5]Browne ML,Van Zutphen AR,Botto LD,et al.Maternal butalbital use and selected defects in the national birth defects prevention study[J].Head,2014,54(1):54-66.
[6]Mateja WA,Nelson DB,Kroelinger CD,et al.The association between maternal alcohol use and smoking in early pregnancy and congenital cardiac defects[J].J Wom Heal,2012,21(1):26-34.
[7]郭佳林,洪向丽,姚明珠.新生儿先天性心脏病危险因素分析[J].中国新生儿科杂志,2010,25(2):76-79.Guo JL,Hong XL,Yao MZ.Analysis of the risk factors of congenital heart diseases[J].Chinese Journal of Neonatology,2010,25(2):76-79.
[8]Alverson CJ,Strickland MJ,Gilboa SM,et al.Maternal smoking and congenital heart defects in the baltimore-washington infant study[J].Ped,2011,127(3):647-653.
[9]李杏肖,李晓红,林秋雄,等.转染isl1基因促进骨髓间充质干细胞向心肌样细胞分化[J].中国病理生理杂志,2012,28(1):131-135.Li XX,Li XH,Lin QX,et al.Isl1 gene transfection promotes BMSCs to differentiate into cardiomyocytes[J].Chinese Journal of Pathophysiology,2012,28(1):131-135.
[10]Ferreira-Martins J,Ogorek B,Cappetta D,et al.Cardiomyogenesis in the developing heart is regulated by c-kit-positive cardiac stem cells[J].Circ Res,2012,110(5):701-715.
[11]Buikema JW,Mady AS,Mittal NV,et al.Wnt/beta-catenin signaling directs the regional expansion of first and second heart field-derived ventricular cardiomyocytes[J].Dev,2013,140(20):4165-4176.
[12]Harmelink C,Peng Y,DeBenedittis P,et al.Myocardial mycn is essential for mouse ventricular wall morphogenesis[J].Dev Biol,2013,373(1):53-63.
[13]Gallagher JM,Komati H,Roy E,et al.Dissociation of cardiogenic and postnatal myocardial activities of gata4[J].Mol Cell Biol,2012,32(12):2214-2223.
[14]Yamak A,Latinkic BV,Dali R,et al.Cyclin d2 is a gata4 cofactor in cardiogenesis[J].Proc Natl Acad Sci USA,2014,111(4):1415-1420.
[15]Maitra M,Schluterman MK,Nichols HA,et al.Interaction of gata4and gata6 with tbx5 is critical for normal cardiac development[J].Dev Biol,2009,326(2):368-377.
[16]Brown CR,Chi X,Garcia-Gras E,et al.The cardiac determination factor,nkx2-5,is activated by mutual cofactors gata-4 and smad1/4 via a novel upstream enhancer[J].J Biol Chem,2004,279(11):10659-10669.
[17]Yang YQ,Li L,Wang J,et al.A novel gata4 loss-of-function mutation associated with congenital ventricular septal defect[J].Ped Car,2012,33(4):539-546.
[18]Zeisberg EM,Ma Q,Juraszek AL,et al.Morphogenesis of the right ventricle requires myocardial expression of gata4[J].J Clin Invest,2005,115(6):1522-1531.
[19]Yang YQ,Wang J,Liu XY,et al.Mutation spectrum of gata4associated with congenital atrial septal defects[J].Arch Med Sci,2013,9(6):976-983.