机构地区:[1]中国人民解放军306医院儿科 [2]北京军区总医院附属八一儿童医院新生儿重症监护中心,100700
出 处:《中华实用儿科临床杂志》2014年第24期1848-1853,共6页Chinese Journal of Applied Clinical Pediatrics
基 金:国家自然科学基金(81170577)
摘 要:目的 探讨孕鼠补充牛磺酸对宫内生长受限(IUGR)胎鼠脑组织蛋白激酶A(PKA)-钙/钙调素依赖性蛋白激酶Ⅱ(CaMKⅡ)/c-fos(PKA-CaMKⅡ/c-fos)信号通路活性的影响.方法 将15只SD孕鼠随机分为对照组、IUGR组、IUGR+孕鼠补充牛磺酸组(牛磺酸组).通过低蛋白饮食法建立IUGR模型,用荧光实时定量(real-time) PCR监测各组胎鼠脑组织PKA、CaMKⅡ、c-fos mRNA的表达水平,Western blot法检测各组胎鼠脑组织PKA、CaMKⅡ、c-fos蛋白表达,免疫组织化学方法检测各组胎鼠脑组织PKA、CaMKⅡ、c-fos阳性细胞数.结果 3组胎鼠脑组织PKA mRNA、CaMKⅡmRNA和c-fos mRNA表达比较差异均有统计学意义(F =7.934,P=0.021;F=5.568,P=0.043;F =7.332,P=0.024).3组胎鼠脑组织PKA蛋白、CaMKⅡ蛋白和c-fos蛋白表达比较差异均有统计学意义(F=57.743,P=0.000;F=163.405,P=0.000;F=160.136,P=0.000).3组胎鼠脑组织PKA阳性细胞、CaMKⅡ阳性细胞和c-fos阳性细胞比较差异亦有统计学意义(F=42.903,P=0.000;F =43.674,P=0.000;F =329.123,P=0.000).与对照组比较,IUGR组胎鼠脑组织PKA、CaMKⅡ和c-fos mRNA、蛋白和阳性细胞数明显降低,差异均有统计学意义(P均<0.05);牛磺酸组与对照组比较无明显差异;牛磺酸组较IUGR组明显增高,差异均有统计学意义(P均<0.05).结论 孕鼠补充牛磺酸可上调IUGR胎鼠脑组织PKA、c-fos和CaMKⅡmRNA及蛋白表达水平,并增加其阳性细胞数,牛磺酸可能通过促进PKA-CaMKⅡ/c-fos信号通路活性而促进IUGR胎鼠脑发育。Objective To explore the effects of prenatal taurine supplementation on the expression of protein kinase A(PKA) and c-fos,Ca^2+/calmodulin-dependent protein kinase Ⅱ (GaMK Ⅱ) in fetal rat brains with intrauterine growth restriction and its significance.Methods Fifteen pregnant Sprague-Dawley rats were randomly divided into 3 groups:the control group,the intrauterine growth restriction (IUGR) group and the IUGR with prenatal taurine supplementation group(the taurine group),with 5 fetal rats in each group.All the fifteen fetal rat brains were detected as following:the expression of PKA,c-fos,CaMK Ⅱ mRNA in fetal rat brains was detected by way of real-time polymerase chain reaction(real-time PCR),while the expression changes of PKA,CaMK Ⅱ,and c-fos protein in fetal rat brains were detected by using Western blot,and the number of PKA,CaMK Ⅱ,c-fos positive cells in fetal rat brains was detected by using immunohistochemistry.Results The control group,the IUGR group and the taurine group:Comparison of the expression of PKA,CaMK Ⅱ,c-fos mRNA among 3 groups were of significant differences(F =7.934,P =0.021 ; F =5.568,P =0.043 ;F =7.332,P =0.024).Comparison of the expression of PKA,CaMK Ⅱ,c-fos protein among the 3 groups were of significant differences(F =57.743,P =0.000 ; F =163.405,P =0.000 ; F =160.136,P =0.000).Comparison of the number of PKA,CaMK Ⅱ,c-fos positive cells among the 3 groups were of significant differences (F =42.903,P =0.000 ;F =329.123,P =0.000 ; F =43.674,P =0.000).Compared with the control group,the expression of mRNA,protein,positive cells of three indicators in IUGR group was less,and the difference was statistically significant (P 〈 0.05).The expression of mRNA,protein,positive cells of three indicators in fetal rat brains of the taurine group were not different from the control group.The expression of mRNA,protein,positive cells of three indicators in fetal rat brains of the taurine group were significantly more than the IUGR group(P
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