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目的:研究维泰醇对小鼠黑色素瘤B16F0细胞糖酵解水平的影响并初步探究其机制。方法:以小鼠黑色素瘤B16F0细胞为研究对象,不同浓度维泰醇处理小鼠黑色素瘤B16F0细胞,另设空白组,采用磺酰罗丹明(SRB)法检测维泰醇对细胞增殖的影响,反向高效液相色谱法检测维泰醇对B16F0细胞ATP含量影响,试剂盒检测B16F0细胞培养液中乳酸含量影响,放射同位素法测定维泰醇对B16F0细胞葡萄糖摄取影响,试剂盒检测维泰醇对B16F0细胞己糖激酶(HK),丙酮酸激酶(PK),乳酸脱氢酶(LDH)活性影响。结果:与空白组比较,维泰醇可浓度依赖性的抑制B16F0细胞ATP的产生和乳酸的释放,减少B16F0细胞对葡萄糖的摄取,浓度依赖性的降低PK和LDH的活性(P<0.05,P<0.01),但对HK的活性无显著性影响。结论:维泰醇可抑制B16F0细胞糖酵解水平,其机制可能与抑制葡萄糖摄取,下调糖酵解的关键酶PK和LDH的活性有关。
OBJECTIVE: To study the effect of vetiol on the glycolytic activity of mouse melanoma B16F0 cells and explore its mechanism. Methods: The mouse melanoma B16F0 cells were treated with different concentrations of vetetinol in mouse B16F0 melanoma cells. A blank group was also established. The effect of vetiverol on cell proliferation was detected by the method of sulforhodamine (SRB) The effects of vetiol on the ATP content of B16F0 cells were determined by reverse-phase high-performance liquid chromatography (RP-HPLC). The contents of lactate in B16F0 cells were assayed by kit, the glucose uptake of B16F0 cells was determined by radioisotope method, On B16F0 cells hexokinase (HK), pyruvate kinase (PK), lactate dehydrogenase (LDH) activity. Results: Compared with the blank group, Viteritol inhibited the ATP production and lactate release in B16F0 cells in a concentration-dependent manner, decreased the uptake of glucose by B16F0 cells and decreased the activity of PK and LDH in a concentration-dependent manner (P <0.05, P <0.01), but no significant effect on the activity of HK. CONCLUSION: Vitermetanol can inhibit the glycolytic activity of B16F0 cells. The mechanism may be related to the inhibition of glucose uptake and down-regulation of PK and LDH activities.