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作 者:黄琰菁[1] 王琳[1] 李赛[1] 盛莉[1] 王海霞[1] 杨生辉[1] HUANG Yanjing;WANG Lin;LI Sai;SHENG Li;WANG Haixia;YANG Shenghui(Department of Oncology,Hainan Provincial People's Hospital,Haikou 570311,Hainan,China)
机构地区:[1]海南省人民医院肿瘤内科
出 处:《中国肿瘤生物治疗杂志》2019年第8期876-881,共6页Chinese Journal of Cancer Biotherapy
基 金:海南省自然科学基金资助项目(No.814315)~~
摘 要:目的:研究罗汉果醇(MO)对肝细胞癌HepG2细胞脂代谢的调控作用及其分子机制。方法:采用油酸(OA)诱导肝细胞癌HepG2细胞脂肪累积,建立脂肪变性细胞模型。运用CCK-8法检测MO对HepG2的细胞毒性,筛选其无明显细胞毒性的实验工作浓度。不同工作浓度MO作用后运用油红O染色法观察模型细胞内脂质累积情况,测定细胞内甘油三酯(TG)、胆固醇(TC)含量。运用高通量转录组测序方法筛选参与脂代谢的关键基因,运用qPCR检测模型及给药细胞SREBP-1c、FASN mRNA的表达、WB法检测p-AMPKα、SREBP-1c、FASN等蛋白的表达水平。结果:运用OA诱导的模型HepG2细胞内脂质大量累积,TG、TC含量显著升高。OA诱导后参与肝癌细胞脂代谢的关键基因SREBP-1c、FASN mRNA表达升高;p-AMPKα蛋白表达降低,SREBP-1c、FASN等蛋白的表达显著升高。工作浓度MO干预后,细胞内脂质累积显著减少、TG和TC含量降低,SREBP-1c、FASN mRNA表达降低,p-AMPKα蛋白表达升高而SREBP-1c、FASN等蛋白的表达明显降低。结论:MO能够通过激活HepG2细胞中AMPK信号通路相关因子SREBP-1c、FASN的表达抑制脂肪酸合成,从而发挥调节脂代谢的作用。Objective:To study the regulatory effect of mogrol(MO) on lipid metabolism of hepatic cancer cells and its molecular mechanism. Methods: Oleic acid(OA) was used to induce fat accumulation in hepatocellular carcinoma HepG2 cells and to establish a steatosis cell model. CCK-8 method was used to detect the cytotoxicity of MO to HepG2 cells, and an experimental working concentration without obvious cytotoxicity of MO was chosen. After being treated with different concentrations of MO, lipid accumulation in the cells was observed by oil red O staining, and the contents of triglyceride(TG) and cholesterol(TC) in the cells were measured. Key genes involving in lipid metabolism were screened out by high-throughput transcriptome sequencing qPCR was used to detect the mRNA expressions of,SREBP-1 c and FASN, while Western Blot was used to detect the protein expressions of p-AMPKα, SREBP-1 c and FASN in cells of model group and treatment group. Results: After OA induction, a large amount of lipids accumulated in HepG2 cells, the contents of TG and TC increased significantly. Three key genes(SREBP-1 c, FASN and p-AMPK α) involving in lipid metabolism of hepatic cancer cells were screened out. After OA induction, the m RNA expressions of SREBP-1 c and FASN increased, the protein expression of p-AMPK α decreased while the protein expressions of SREBP-1 c, FASN and other proteins increased significantly.After intervention with working concentration of MO, intracellular lipid accumulation, contents of TG and TC, mRNA expressions of SREBP-1 c, FASN and protein expressions of SREBP-1 c, FASN decreased significantly, while the expression of p-AMPKα increased.Conclusion: Mogrol can inhibit the synthesis of fatty acids by activating the expression level of AMPK signaling pathway related factors SREBP-1 c and FASN, so as to play the role of regulating lipid metabolism.
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