表1三组大鼠窦房结周围界嵴处和腔静脉窦处的Cx43荧光强度(略)
aP<0.05 vs幼年;bP<0.01 vs幼年; cP<0.05 vs成年.
3讨论
窦房结包埋在右心房内,结内兴奋如何传导到心房,而起搏点如何免受心房肌的超极化抑制,这与细胞间电信号传导有很大的关系. 从窦房结中央区到周围心房肌细胞间缝隙连接蛋白分布目前有两种观点,Joyner等[5]认为从窦房结中央区到周围心房肌细胞间耦合逐渐增多,这保证窦房结能正常启动心房除极,然而更多学者发现兔和小鼠窦房结中央区到周围心房肌细胞间耦合无逐渐变化规律,而是表达Cx43和Cx40的心房肌细胞束延伸到主要由表达Cx45的P细胞构成的窦房结中央区,又因为Cx43和Cx40的电导大于Cx45[68],因此,起源于窦房结中央区的冲动沿着这些心房肌束顺利地传导到心房,而不能逆向传导.
不同种属窦房结及周围心房组织缝隙连接蛋白分布差异很大,目前研究未发现大鼠窦房结表达Cx40,心房是否表达Cx40各家报道不一[1-2],窦房结中是否有Cx45表达未见报道,Cx45在心房肌表达弱[2-3],而Cx43在窦房结不表达,而在心房肌强表达[2]. 因此,Cx43是大鼠窦房结周边区的主要蛋白,主要负责冲动由窦房结传导到心房[4].
Jones等[9]用免疫荧光法研究豚鼠窦房结缝隙连接蛋白的增龄性改变,发现Cx43在豚鼠出生时存在于整个窦房结和心房肌,1月龄时,SAN中心区即为阴性,以后,随年龄增长Cx43阴性区域逐增大,Western Blot法发现随年龄增加Cx43表达量减少,Cx40, Cx45表达不变. 我们对大鼠窦房结周边区Cx43荧光强度进行研究,发现在不同年龄段大鼠中Cx43在表达HCN4的窦房结区为阴性,未发现Cx43阳性的心房肌延伸到HCN4阳性的窦房结区,窦房结周边区Cx43表达随年龄增长逐渐减少. 免疫荧光测量的阀值是每个缝隙连接处要有大于90个缝隙连接蛋白通道[4],Cx45表达于很多种属动物窦房结,而在大鼠窦房结未见报道,可能是Cx45表达低于免疫荧光测量的阀值,同样作者未发现Cx43阳性的心房肌延伸到HCN4阳性的窦房结区,是否为Cx43表达低于免疫荧光测量的阀值,这需要寻找更精确的实验方法来判断.
Kuga等[10]观察到人SACT随年龄增长而延长,本实验也发现大鼠SACT随年龄增长亦逐渐延长. 电生理研究发现随年龄增长,从SAN中心区到外周区,传导时间延长,传导距离加大,传导速度减慢[9]. 大鼠窦房结周围界嵴处和心房处的Cx43对冲动传导到心房有重要作用,本研究发现,随年龄增长大鼠窦房结周围界嵴处和固有心房处的Cx43表达下降,这可能与窦房结冲动传导到心房速度减慢有关,可能是窦房结功能随增龄而下降的原因之一,但这种现象的确切机制尚需进一步探讨.
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