对正常大鼠心肌细胞连接立体结构及连接蛋白表达增龄变化的研究

发布时间：2018-02-27 23:08

本文关键词： Cx43 Cx40 N-cadherin α-catenin 缝隙连接增龄扫描电镜激光共聚焦显微镜　出处：《河北医科大学》2005年硕士论文　论文类型：学位论文

【摘要】：目的:本研究应用HE 染色、免疫组织化学技术、激光共聚焦显微镜技术及消化扫描电镜技术,对胚胎、新生、幼年、成年、中年及老年正常大鼠左心室心肌细胞闰盘的立体构筑和连接蛋白Connexin43(Cx43)、Connexin(Cx40)、N-cadherin、α-catenin 含量、分布的增龄变化进行了研究,以期为人类心脏发育相关研究提供生物学资料。方法:本实验选取SD 大鼠胎鼠5 只(孕20 天)和出生后雄性SD 大鼠,按年龄分为新生组(出生3～5 天),幼年组(出生20～25 天),体重25±4g;青年组(出生3～4 月),体重250±10g;中年组(出生6～8 月),体重350±10g和老年组(出生13～15 月),体重450±10g,每组各5 只。1、选取上述6 组动物的心脏经主动脉灌流4%多聚甲醛后,常规石蜡包埋,横断面连续系列切片,片厚5μm。HE 染色、免疫组织化学技术结果结合图像分析系统分别检测大鼠心肌Cx43、Cx40、N-cadherin、α-catenin 的表达情况,并计量阳性表达的平均光密度值(OD/um~2)。2、激光扫描共聚焦显微镜技术选取青年组动物,取材方法同前,采用Cx43 标红色荧光、Cx40 标绿色荧光进行荧光双重标记。3、扫描电镜技术选取青年、中年和老年组,心脏样品取心外膜侧0.5×0.5×0.3 cm~3 大小,经6N 氢氧化钠水溶液620C 消化10min,超声波处理1min,常规脱水、干燥、镀膜后进行扫
[Abstract]:Objective: to study the effects of HE staining, immunohistochemistry, laser confocal microscopy and digestive scanning electron microscopy on embryo, neonate, infancy and adulthood. The stereoarchitecture of intercalated disc of left ventricular cardiomyocytes in middle-aged and aged normal rats and the changes of Connexin43Cx43Cx43Cx40Cx40, N-cadherin, 伪 -catenin content and distribution were studied in order to provide biological data for the study of human heart development. Methods: in this experiment, 5 SD rats (20 days pregnant) and 5 postnatal male SD rats were selected. According to age, they were divided into newborn group (35 days old), young group (20 ~ 25 days old, weight 25 卤4 g), youth group (3 ~ 4 months old, weight 250 卤10 g), middle age group (68 months old, 350 卤10 g) and elderly group (13 ~ 15 months old, 450 卤10 g, 5 rats in each group). The hearts of the above 6 groups were perfused with 4% paraformaldehyde through aorta. The expression of Cx43 Cx40 N-cadherin and 伪 -catenin in rat myocardium was detected by immunohistochemical technique and image analysis system. The average optical density of the positive expression was measured. The young animals were selected by laser scanning confocal microscopy (LSCM). The method was the same as that before. The green fluorescence labeled with Cx43 was used to carry out the double labeling. 3, and the young people were selected by scanning electron microscopy. In the middle-aged and elderly groups, the epicardial side of the heart was taken as 0.5 脳 0. 5 脳 0. 3 cm~3, digested by 620C sodium hydroxide solution for 10 minutes, then treated with ultrasound for 1 min, then dehydrated, dried, and scanned after coating.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R33

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