摘要
目的探讨刺激新生大鼠心肌细胞(NRCMs)整联蛋白增加某些大分子物质的摄取及其机制。方法以100～300mg/L甘氨酸-精氨酸-甘氨酸-天冬酰胺-色氨酸序列(RGD)刺激NRCMs整联蛋白,同时加入不同浓度德克萨斯红标记的卵蛋白(OTR)、右旋糖苷(DTR),孵育8h、16h、24h,应用免疫荧光技术研究心肌细胞对OTR、DTR的摄取及Dysferlin面积。结果 RGD增加NRCMs对OTR、DTR的摄取(与对照组比较P<0.001,P<0.05),并与孵育时间呈正相关(P<0.001),OTR的摄取与其浓度呈正相关(P<0.001)。RGD组Dysferlin面积为无RGD组的3倍。RGD刺激非搏动的心肌细胞,不增加OTR、DTR的摄取,亦不增加Dysferlin面积。结论 RGD刺激增加NRCMs对OTR、DTR的摄取。Dysferlin参与RGD诱导的心肌细胞膜损伤的修复过程。RGD诱导的心肌细胞膜损伤与细胞的收缩活动相关。
Objective To test whether stimulation of integrins on neonatal rat cardiomyocytes （NRCMs） would favor uptake of macromoleeules. Methods NRCMs were treated with Gly-Arg-Gly-Asp-Ser （ RGD, 100-300 mg/L） to stimulate integrins, or phosphate-buffered saline, in the presence of ovalbumin-texas red （OTR, 45 kD） or dextran-texas red （DTR, 70 kD）. After 8 hours, 16 hours, 24 hours, uptake of red label as well as Dysferlin immunoreaetivity were quantified by fluorescence microscopy. Results Uptake of OTR and DTR by NRCMs was intensified by RGD （ for trend P 〈 0. 001 and P 〈 0.05, respectively） and was correlated with duration of incubation （P 〈 0. 001 for both）. RGD-induced uptake of OTR by NRCMs was correlated with OTR concentration （P 〈 0. 001 ）. The Dysferlin-positive areas in RGD- treated NRCMs were about 3-fold larger than in NRCMs incubated without RGD （P 〈 0. 001 ）. However, in non-beating NRCMs RGD did not induce larger dysferlin-positive areas, nor did RGD stimulate uptake of OTR. Conclusion In NRCMs stimulation of integrins caused uptake of labeled macromolecules. Dysferlin may be involved in membrane repair following RGD-indueed membrane wounding. RGD-induced membrane wounding is closely associated with contractile behavior of the NRCMs.
出处
《解剖学报》
CAS
CSCD
北大核心
2010年第3期473-476,共4页
Acta Anatomica Sinica
基金
吉林省科技发展计划项目(200705405)
