清燥救肺汤对荷Lewis小鼠肺癌细胞中ACO1,ACL,FFA,CHO的影响

目的：观察清燥救肺汤对荷Lewis小鼠肺癌细胞顺乌头酸酶(Aconitase closing odor,ACO1)、ATP柠檬酸裂解酶(ATP citrate lyase,ACL)、游离脂肪酸(Free fatty acids,FFA)、胆固醇(Cholesterol,CHO)含量的影响。方法：C57BL/6J雄性小鼠,随机分为5组,分别为清燥救肺汤高剂量组、中剂量组、低剂量组,模型组,环磷酰胺(Cyclophosphamide,CTX)组,每组10只,共50只。于小鼠右腋下注射Lewis肺癌细胞建立肺癌荷瘤模型,清燥救肺汤低、中、高剂量组剂量分别为2.75 ,5.5,11g.kg_^-1.d_^-1,先灌胃给药2周然后造模,CTX组以50 mg.kg_^-1.(2d)_^-1进行腹腔注射给药,模型组用等体积的生理盐水进行灌胃给药,接种后继续灌胃给药2周,处死小鼠取瘤组织。酶联免疫吸附测定(Enzyme-linkedimmunesorbent assay,ELISA)检测FFA、CHO的含量;实时荧光定量聚合酶链式反应(Real-time PCR)检测ACL mRNA的表达;蛋白质免疫印记法(Western blot)检测ACO1蛋白表达。结果：与模型组比较,CTX组及清燥救肺汤高、中、低剂量组中肺癌细胞ACO1蛋白表达,ACL mRNA的表达及FFA、CHO含量显著降低(P＜0.05,P＜0.01)。结论：清燥救肺汤可有效抑制荷Lewis肺癌细胞增殖,减少肺癌细胞能量生成,ACO1、ACL、FFA、CHO可能是其药效作用靶点。

Effects of Qingzao Jiufei Decoction on ACO1, ACL, FFA and CHO in Lewis-bearing mice lung cancer cells

Objective:To observe the effects of Qingzao Jiufei Decoction on the contents of Aconitase closing odor (ACO1), ATP citrate lyase (ACL), free fatty acids (FFA) and cholesterol (CHO) in lung cancer cells of Lewis mice. Method: C57BL/6J male mice were randomly divided into Qingzao Jiufei Decoction high, medium and low dose groups, model group and cyclophosphamide (CTX) group,with 10 mice in each group,a total of 50. Lewis lung cancer cells were injected into the right axillary of all mice to establish lung cancer bearing model. The dosage of Qingzao Jiufei Decoction in low , middle and high dose groups was 2.75, 5.5 and 11 g.kg_^-1.d_^-1.First intragastric administration for 2 weeks and the model was established. The mice in CTX group were administrated by intraperitoneal injection of 50 mg.kg_^-1.2d_^-1, and the mice in model group were administrated by intragastric administration of equal volume normal saline after inoculation.Continued intragastric administration for 2 weeks after inoculation, mice were executed to get tumor tissues.The contents of FFA and CHO were detected by enzyme-linked immunosorbent assay (ELISA); the expression of ACL gene was detected by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR);and the expression of ACO1 protein was detected by Western blot. Result:As compared with model group, the expression of ACO1 protein, the expression of ACL gene and the contents of FFA and CHO in lung cancer cells in high, medium and low dose groups and CTX group of Qingzao Jiufei Decoction were significantly decreased(P<0.05,P<0.01). Conclusion:Qingzao Jiufei Decoction can effectively inhibit the proliferation of Lewis lung cancer cells and reduce the energy production of lung cancer cells. ACO1, ACL, FFA and CHO may be the targets of its pharmacodynamics.