目的探讨驱动蛋白KIF4A对巨噬细胞不同亚群中血管生成相关因子表达的影响。方法 THP-1细胞在PM A及不同人重组细胞因子的外界刺激下分别分化为M0、M1、M2细胞,采用ELISA技术检测此3种巨噬细胞亚群中血管生成相关因子的表达水平。利用siRNA转染敲除M0、M1、M2细胞中KIF4A的表达,采用实时定量PCR检测敲除后血管生成相关因子的表达水平。结果巨噬细胞M0上清中可检测到LIF、VEGF、s VEGFR1及TIM P1,而s VEGFR2、Flt-3、Leptin、LeptinR、CD40L均不表达。其中LIF、VEGF、TIM P1在M0向M1、M2分化过程中,表达差异均无统计学意义(P〉0.05);s VEGFR1在M0向M1分化过程中表达差异无统计学意义(P〉0.05),向M2分化过程中表达显著上调(P〈0.01)。转染KIF4A siRNA后,M0与M1细胞中LIF、s VEGFR1、VEGF、TIM P1的表达差异均无统计学意义(P〉0.05);M2细胞中LIF、VEGF、TIM P1表达差异均无统计学意义(P〉0.05),但s VEGFR1表达水平明显降低(P〈0.05)。结论 M2细胞s VEGFR1表达水平显著上调;KIF4A参与M2细胞中s VEGFR1的表达。
Objective To explore the effect of KIF4 A on expressions of angiogenesis-associated factors in M0,M1 and M2 cells. Methods THP-1 cells were differentiated into M0,M1 and M2 cells under the treatment of PMA and different recombination human cytokines in vitro. The expressions of several angiogenesis-associated factors in these three macrophage subpopulations were analyzed by ELISA. KIF4 A siRNA was transfected into M0,M1 and M2 cells,and the levels of angiogenesis-associated factors after the transfection were analyzed by qRT-PCR. Results LIF,VEGF,s VEGFR1 and TIMP1 were detected from supernatant of M0 cells,whereas the expressions of s VEGFR2,Flt-3,Leptin,LeptinR and CD40 L were not detected. The expressions of LIF,VEGF and TIMP1 did not significantly change during the differentiation from M0 to M1 or M2 cells( P〉0. 05). The expression of s VEGFR1 was also constant in M0 and M1 cells( P〉0. 05),but its expression was significantly up-regulated in M2 cells( P〈0. 01). After the transfection of KIF4 A siRNA,the levels of LIF,s VEGFR1,VEGF and TIMP1 did not significantly change in M0 and M1 cells,and LIF,VEGF and TIMP1 in M2 cells( P〉0. 05). However,s VEGFR1 expression in M2 cells was significantly decreased after KIF4 A siRNA transfection( P〈0. 05). Conclusion M2 cells exhibite higher s VEGFR1 expression and KIF4 A is involved in s VEGFR1 expression in M2 cells.