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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 investigate the effect of kinesin KIF4A on the expression of angiogenesis-related factors in different subpopulations of macrophages. Methods THP-1 cells differentiated into M0, M1 and M2 cells under the external stimuli of PM A and different human recombinant cytokines. The expression of angiogenesis-related factors in these three macrophage subsets was detected by ELISA. The expression of KIF4A in M0, M1 and M2 cells was knocked down by siRNA transfection, and the expression level of angiogenesis-related factors after knockdown was detected by real-time quantitative PCR. Results LIF, VEGF, s VEGFR1 and TIMP1 were detected in M0 supernatant of macrophages, while none of s VEGFR2, Flt-3, Leptin, LeptinR and CD40L were detected. There was no significant difference in the expression of LIF, VEGF and TIMP1 between M0 and M1 and M2 (P> 0.05). There was no significant difference in VEGFR1 expression between M0 and M1 (P> 0.05) , Up-regulated to M2 differentiation (P <0.01). The expression of LIF, s VEGFR1, VEGF and TIM P1 in M0 and M1 cells was not significantly different after transfected with KIF4A siRNA (P> 0.05). There was no significant difference in the expressions of LIF, VEGF and TIM P1 (P> 0.05), but the expression of s VEGFR1 was significantly decreased (P <0.05). Conclusion The expression of VEGFR1 in M2 cells was significantly up-regulated. KIF4A was involved in the expression of VEGFR1 in M2 cells.