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目的探讨羰基-氰-对-三氟甲氧基本腙(FCCP)对脂肪酸结合蛋白3(FABP3)基因表达沉默的P19细胞线粒体功能的影响。方法构建FABP3表达沉默载体(A组)及相应的空载体(B组),包装病毒后感染P19细胞,建立稳定转染FABP3沉默的P19细胞株,采用实时定量PCR检测干扰效率。给予FCCP 2.5μmol/L(C组)、5μmol/L(D组)干预FABP3沉默的P19细胞,采用荧光素酶化学发光法检测细胞ATP含量,流式细胞术观察线粒体膜电位的变化。结果 B组FABP3基因表达量较A组下调(0.50±0.08vs.2.23±0.42)(P<0.05)。与A、B组相比,C、D组细胞ATP生成减少(2.47±0.58、1.05±0.13vs.0.09±0.03、0.12±0.02)(P<0.05),而细胞线粒体膜电位升高(358.36±12.23、389.66±12.13vs.437.35±9.27、473.21±19.65)(P<0.05)。结论FABP3基因在维持P19细胞线粒体功能中发挥重要作用。
Objective To investigate the effects of carbonyl-cyano-p-trifluoromethoxyhydrazone (FCCP) on the mitochondrial function of P19 cells with silenced FABP3 gene expression. Methods The vector of FABP3 silencing vector (group A) and corresponding empty vector (group B) were constructed and infected with P19 cells after virus infection. The P19 cell line stably transfected with FABP3 was established and the interference efficiency was detected by real - time PCR. FCM 2.5μmol / L (group C) and 5μmol / L (group D) were used to intervene the FABP3-silenced P19 cells. The ATP content of the cells was detected by luciferase chemiluminescence and the change of mitochondrial membrane potential was observed by flow cytometry. Results The expression of FABP3 gene in group B was significantly lower than that in group A (0.50 ± 0.08 vs.2.23 ± 0.42) (P <0.05). Compared with group A and group B, the ATP production in C and D groups decreased (2.47 ± 0.58, 1.05 ± 0.13 vs. 0.09 ± 0.03, 0.12 ± 0.02, P <0.05), while the mitochondrial membrane potential increased (358.36 ± 12.23, 389.66 ± 12.13 vs. 437.35 ± 9.27, 473.21 ± 19.65) (P <0.05). Conclusion FABP3 gene plays an important role in maintaining the mitochondrial function of P19 cells.