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肝部分切除术后Tmub1蛋白与securin蛋白的相互作用研究
作者:赵晓彪 李光耀 刘孟刚 范霞 陈平 
单位:400042 重庆 第三军医大学大坪医院野战外科研究所肝胆外科(赵晓彪、李光耀、刘孟刚、陈平) 一室(范霞) 
关键词:Tmub1蛋白 securin蛋白 肝再生 肝切除术 
分类号:R657.3
出版年,卷(期):页码:2014,39(11):864-867
摘要:
目的 探讨肝部分切除术后Tmub1蛋白与securin蛋白的相互作用。方法 SD大鼠30只,随机分为肝切除术后0、24、48h组,每组10只。0h组行肝切除术后直接灌注肝脏并取原代肝细胞,后两组分别于术后24、48h灌注肝脏提取原代肝细胞。每组肝细胞中一部分行原代肝细胞培养,并通过免疫荧光及激光共聚焦实验检测Tmub1蛋白与securin蛋白的共定位情况;另外一部分肝细胞经裂解后提取蛋白,行免疫共沉淀及Western blotting检测Tmub1/securin共沉淀蛋白的表达。结果 激光共聚焦实验结果显示,肝切除术后0h细胞核内几乎没有Tmub1和securin蛋白表达,术后24h及48h Tmub1与securin蛋白大量共定位于细胞核内;免疫共沉淀及Western blotting检测结果显示,术后0h Tmub1/securin蛋白条带较弱,术后24~48h明显增强(P<0.01),但24h与48h之间差异无统计学意义(P>0.05)。结论 Tmub1在正常肝细胞中几乎不与securin结合,但在肝切除术后从胞质进入细胞核,与securin蛋白相互结合,从而抑制肝细胞的增殖。
Objective To investigate the interaction of Tmub1 and securin protein after partial hepatectomy. Methods A total of 30 adult male SD rats were randomly divided into 3 groups (10 each) according to the time after hepatectomy, i.e, 0, 24 and 48h. Partial hepatectomy was performed in the rats in the first group, and then the primary hepatocytes were directly harvested by liver perfusion. Partial hepatectomy was also performed in the rest two groups at postoperative 24 and 48 h. Then the primary hepatocytes were also harvested respectively after liver perfusion. One part of primary hepatocytes was cultured for one night and for the observation of the colocalization of Tmub1 and securin protein with immunofluorescence. The other part of hepatocytes were used for co-immunoprecipitation and Western blotting to detect the expression of protein Tmub1/securin. Results Immunofluorescence and confocal laser scanning microscopy showed that there were almost no Tmub1 and securin proteins in the nucleus 0 h after liver resection, while a great amount of Tmub1/securin proteins were found to be co-localized in the nucleus 24 and 48 h. Co-immunoprecipitation and Western blotting showed that the expression of Tmub1/securing protein significantly enhanced in 24-48 h after operation compared with that at 0 h (P<0.01), while the difference between 24 h and 48 h showed no statistically significant difference (P>0.05). Conclusion Tmub1 hardly combines with securin protein in normal hepatocytes, but it combines with securin protein when it migrates from cytoplasm into nucleus after hepatectomy, thus inhibiting the proliferation process of hepatocytes.
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