摘要
目的探讨缝隙连接阻断剂对大鼠血管平滑肌细胞表型转化的影响。方法采用贴块法培养大鼠主动脉平滑肌细胞,细胞免疫荧光染色法检测大鼠主动脉平滑肌细胞缝隙连接蛋白43的表达,荧光漂白后恢复技术检测缝隙连接介导的细胞间通讯。四唑盐比色法测定细胞增殖能力,逆转录聚合酶链反应法检测平滑肌α-肌动蛋白的表达,并观察缝隙连接特异性阻断剂18α-甘草次酸对上述指标的影响。结果大鼠血管平滑肌细胞体外培养3天后可表达缝隙连接蛋白43。荧光物质能够通过缝隙连接在相邻细胞间进行传递,孤立细胞荧光恢复率显著低于相邻细胞(7.30%±0.58%比80.61%±6.57%,P<0.01);而18α-甘草次酸能够抑制缝隙连接介导的细胞间通讯,18α-甘草次酸组荧光恢复率显著低于对照组(61.43%±7.62%比80.61%±6.57%,P<0.05)。18α-甘草次酸可抑制血管平滑肌细胞增殖,并且可促进平滑肌α-肌动蛋白信使核糖核酸的表达。结论缝隙连接阻断剂可促进大鼠血管平滑肌细胞由合成型向收缩型转化,提示缝隙连接在调节血管平滑肌细胞表型转化过程起一定作用。
Ahn To explore the effect of the blocker of gap junction on phenotypic transition in cultured rat vascular smooth muscle cells. Methods Rat aortic smooth muscle cells （SMC） were cultured by explanted rat aortic wall tissue. Cell immunofluoreseence staining was applied to detect the expressions of connexin （Cx）43 in rat aortic SMC. Fluorescence redistribution after photobleaching （FRAP） was used to measure the communications between cells via gap junctions. MTT and RT-PCR were hired to measure the proliferative capability of rat aortic SMC and the expression of smooth muscle （SM） α-actin respectively. Meanwhile, 18α-glycyrrhetinic acid （18α-GA）, a specific blocker of gap junction, was administered to observe its effect on the contents above. Results At 3rd day after cultured, Cx43 was expressed in rat aortic SMC. Fluorescent dye could only be transferred between conjugated cells, and mean fluorescence recovery rate in isolated cells were significantly lower compared with that in conjugated cells （7.30% ±0.58% vs 80.61% ±6.57%, P〈0.01）. Compared to control group, fluorescence recovery rate in 18α-GA group were significant lower （61.43% ± 7.62% vs 80.61% ± 6.57%, P 〈 0.05）. Therefore, 18α-GA could inhibit dye transfer between conjugated cells. 18α-GA could also inhibit the proliferation of rat aortic SMC and promote the expression of SM a-actin in cultured rat aortic SMC. Condusions The blocker of gap junction could prt,mote the phenotypic transition rat aortic SMC from the synthetic to the contraetile phenotype, which suggested that gap junction played a role on the phenotypic modulation in cultured rat aortic MSC.
出处
《中国动脉硬化杂志》
CAS
CSCD
2008年第2期85-88,共4页
Chinese Journal of Arteriosclerosis
基金
国家自然科学基金(30570765
30700889)
