固脾消积饮对肝星状细胞共培养条件下HepG2细胞氨基酸代谢和能量代谢的影响  被引量:6

Effects of Gupi Xiaoji Decoction on Amino Acid Metabolism and Energy Metabolism of HepG2 Cells under Co-culture of Hepatoma Stellate Cells

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作  者:柳卓 田雪飞[2] 谭小宁[3] 郜文辉[2] 翦慧颖[2] 李可心 周琳 张振 曾普华 LIU Zhuo;TIAN Xuefei;TAN Xiaoning;GAO Wenhui;JIAN Huiying;LI Kexin;ZHOU Lin;ZHANG Zhen;ZENG Puhua(Hunan Academy of Traditional Chinese Medicine,Changsha 412000,China;Hunan University of Chinese Medicine,Changsha 412006,China;Affiliated Hospital of Hunan Academy of Traditional Chinese Medicine,Changsha 412000,China)

机构地区:[1]湖南省中医药研究院,湖南长沙412000 [2]湖南中医药大学,湖南长沙412006 [3]湖南省中医药研究院附属医院,湖南长沙412000

出  处:《中国中医药信息杂志》2023年第4期94-99,共6页Chinese Journal of Information on Traditional Chinese Medicine

基  金:国家自然科学基金(82074425);湖南省重点研发计划(2021SK2006);湖南省自然科学基金(2021JJ30417);湖南省中医药管理局青年项目(2021178);抗肿瘤中药创制技术湖南省工程研究中心项目(2016年);国家中医药管理局青年岐黄学者项目(2020年)。

摘  要:目的 观察固脾消积饮对人肝星状细胞LX-2共培养条件下肝癌细胞HepG2氨基酸代谢和能量代谢的影响。方法 建立HepG2细胞和LX-2细胞共培养体系,与单独培养的HepG2细胞同时设置对照组、空白血清组、中药血清组(固脾消积饮含药血清)和阳性药组(顺铂)。干预24 h后,CCK8法检测细胞增殖活力,DAPI染色观察细胞状态,ELISA检测细胞上清液三酰甘油(TG)、总胆固醇(TC)、丙酮酸和葡萄糖含量,试剂盒检测细胞丙二醛(MDA)含量和糖酵解速率,高效液相色谱法检测细胞上清液氨基酸含量,Western blot检测腺苷酸活化蛋白激酶(AMPK)α表达。结果 与同一培养体系对照组比较,10%、20%中药血清组及阳性药组单独培养和共培养HepG2细胞增殖活力降低,上清液TG、TC、葡萄糖含量减少,丙酮酸含量增加,细胞糖酵解速率降低,MDA含量增加,17种氨基酸含量减少(P<0.05,P<0.01)。Western blot结果显示,10%中药血清能升高单独培养和共培养HepG2细胞p-AMPKα/AMPKα比值。结论 固脾消积饮能调节肝星状细胞共培养条件下HepG2细胞氨基酸代谢和能量代谢,其机制与激活p-AMPKα通路有关。Objective To observe the effects of Gupi Xiaoji Decoction on amino acid metabolism and energy metabolism of HepG2 cells under co-culture of human hepatoma stellate LX-2 cells.Methods A co-culture system of HepG2 cells and LX-2 cells was established.The control group,blank serum group,TCM serum group(Gupi Xiaoji Decoction containing serum) and positive drug group(cisplatin) were set simultaneously with HepG2 cells cultured alone.After 24 hours of intervention,the proliferation activity of HepG2 was detected by CCK8 method and apoptosis was observed by DAPI staining,the contents of triacylglycerol(TG),cholesterol(TC),pyruvate and glucose in cell supernatant were detected by ELISA,kit was used to detect cellular malondialdehyde(MDA) content and glycolysis rate,the content of amino acids in cell supernatant was detected by HPLC,the expression of adenylateactivated protein kinase(AMPK)α was detected by Western blot.Results Compared with the control group of the same culture system,the proliferation activity of HepG2 cells cultured alone and co-cultured in the 10%,20% TCM serum group and the positive drug group decreased,the contents of TG,TC and glucose in supernatant reduced,the content of pyruvate increased,the glycolysis rate of cell decreased,the content of MDA increased,and the content of 17 amino acids decreased,all with statistical significance(P<0.05,P<0.01).Western blot results showed that 10% TCM serum could increase the ratio of p-AMPKα/AMPKα in HepG2 cells cultured alone and co-cultured.Conclusion Gupi Xiaoji Decoction can regulate the amino acid metabolism and energy metabolism of HepG2 cells under the co-culture condition of hepatic stellate cells,and its mechanism is related to the activation of p-AMPKα pathway.

关 键 词:固脾消积饮 HEPG2 LX-2 氨基酸代谢 能量代谢 

分 类 号:R285.5[医药卫生—中药学]

 

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