3种活血化瘀中药复方含药血清对神经胶质瘤U251细胞JAK/STAT信号通路的影响
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篇名: 3种活血化瘀中药复方含药血清对神经胶质瘤U251细胞JAK/STAT信号通路的影响
TITLE:
摘要: 目的:评价3种活血化瘀中药复方含药血清对神经胶质瘤U251细胞侵袭行为及Janus激酶(JAK)/信号转导及转录激活因子(STAT)通路的影响。方法:将大鼠随机分为生理盐水组(5 mL/kg)、桃红四物汤组(5.7 g/kg)、血府逐瘀汤组(8.5 g/kg)和抵挡汤组(2.8 g/kg),以生药计,每天ig给药1次,连续10 d,末次给药2 h后制备10%含药血清培养液。以10%含药血清培养液干预U251细胞1周后,采用Transwell法检测细胞侵袭率,Western blot法检测细胞中金属基质蛋白酶2(MMP-2)、MMP-9、磷酸化JAK2(p-JAK2)、磷酸化STAT3(p-STAT3)蛋白表达,实时荧光定量聚合酶链式反应法检测细胞中MMP-2、MMP-9 mRNA表达。结果:与空白血清比较,血府逐瘀汤含药血清能降低细胞侵袭率(P<0.05),下调细胞中MMP-2、MMP-9 mRNA及其蛋白表达以及细胞中p-JAK2、p-STAT3蛋白表达(P<0.05或P<0.01);而桃红四物汤含药血清和抵挡汤含药血清作用后上述指标差异无统计学意义 (P>0.05)。结论:在3种活血化瘀中药复方中,血府逐瘀汤具有显著抑制U251细胞侵袭的能力;其机制可能与抑制JAK2/STAT3信号通路激活,下调MMP-2、MMP-9基因及蛋白表达有关。
ABSTRACT: OBJECTIVE: To evaluate the effects of 3 kinds of serum containing blood-activating and stasis-eliminating TCM compound formulas on aggressive behavior, Janus kinase (JAK)/signal transduction and transcriptional activator (STAT) pathway of glioma U251 cells. METHODS: Rats were randomly divided into normal saline group (5 mL/kg), Taohong Siwu decoction group (5.7 g/kg), Xuefu Zhuyu decoction (8.5 g/kg) and Didang decoction (2.8 g/kg), calculated by crude drug, intragastrically administrated once a day, for 10 d. 10% drug-containing serum culture medium was prepared after 2 h of last administration. After 10% drug-containing serum culture medium intervening U251 cells for 1 week, Transwell method was conducted to detect the cell invasion rate, Western blot was adopted to detect the metal matrix protease 2 (MMP-2), MMP-9, phosphorylated JAK2 (p-JAK2), phosphorylated STAT3 (p-STAT3) protein expression; and real-time fluorescence quantitative polymerase chain reaction method was used to detect MMP-2, MMP-9 mRNA expression. RESULTS: Compared with blank serum, Xuefu Zhuyu decoction drug-containing serum can reduce cell invasion rate (P<0.05), decrease the MMP-2, MMP-9 mRNA and its protein expression, and p-JAK2, p-STAT3 protein expression in cells (P<0.05 or P<0.01), while there was no significant difference in above-mentioned indexes in Taohong Siwu decoction drug-containing serum group and Didang drug-containing serum group (P>0.05). CONCLUSIONS: In the 3 kinds of blood-activating and stasis-eliminating TCM compound formulas, Xuefu Zhuyu decoction shows significant invasive effect on inhibiting U251 cells; the mechanism may be related to inhibiting the activation of JAK2/STAT3 signal pathway and decreasing MMP-2, MMP-9 gene and protein expressions.
期刊: 2017年第28卷第16期
作者: 刘建民,黄良文,朱旭红,胡鹏,袁淮涛
AUTHORS: LIU Jianmin,HUANG Liangwen,ZHU Xuhong,HU Peng,YUAN Huaitao
关键字: 活血化瘀;侵袭;Janus激酶/信号转导及转录激活因子信号通路;金属基质蛋白酶;胶质瘤U251细胞
KEYWORDS: Blood-activating and stasis-eliminating; Invasion; Janus kinase/signal transduction and transcriptional activator pathway; Metal matrix protease; Glioma U251 cells
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