【摘要】 目的探讨凝血酶是否上调体外培养的大鼠星形胶质细胞(As)核转录因子(NF-κB)的表达,及其对细胞增殖的影响。方法对新生Wistar鼠大脑As体外培养并纯化,经不同浓度(0.1、1、10、100U/ml)凝血酶干预后,用免疫细胞化学染色和逆转录多聚酶链反应(RT-PCR)检测培养各组AsNF-κBmRAN和蛋白的表达;用化学反应法测定培养细胞上清液中乳酸脱氢酶活性(LDH);用MTT法观察不同浓度凝血酶对细胞生长活性的影响;用流式细胞术分析凝血酶对细胞周期和凋亡的影响。结果与未用凝血酶的对照组比较,各种浓度凝血酶均可引起AsNF-κBmRNA水平增加;凝血酶(1、10、100U/ml)还引起了细胞活性的明显下降;同时导致了培养上清液中LDH活性增高;另外,浓度为100U/ml的凝血酶,可使细胞阻滞在G2/M期的比例增加。结论凝血酶能上调体外培养As的NF-κB的表达,降低细胞生长活性,影响细胞生长周期。更多还原

【Abstract】 Objective To determine whether thrombin can up-regulate the expression of NF-κB in cultured cortical astrocyte and the effect of thrombin on cell proliferation. Methods Primary astrocyte was dissociated from new-born Wistar rats and then purified. The purified astrocytes were treated by thrombin at different concentrations (0.1, 1, 10, 100 U/ml). Immunocytochemistry dyeing and RT-PCR were used to measure the protein and gene expression of NF-κB in cultured astrocytes. The activity of lactate dehydrogenate of cultured liquid was determined by chemistry reaction. The viability of astrocytes were measured by MTT. Flow cytometry was used to determine the proliferation and apoptosis of cultured astrocytes. Results Compared with the control group without thrombin treatment, various concentrations of thrombin increased gene expression of NF-κB in cultured astrocytes, the viability of astrocytes decreased while the activity of LDH in the supernatant increased after 0.1, 1, 10, 100 U/ml thrombin treatment. 100u/ml thrombin treatment led the proliferated cycle of a higher percentage of astrocytes to be blocked at the stage G2/M. Conclusions Thrombin can up-regulate the expression NF-κB in cultured cortical astrocytes, decreased viability of astrocytes and adversely affect cycle of the proliferative astrocytes at the stage G2/M.更多还原