银杏叶提取物含药血清间接抑制肝癌细胞株恶性进展的机制  

Study on the Indirect Inhibition of Hepatocellular Carcinoma Cell Lines by the Decrease of Platelet Activating Factor in Ginkgo Biloba Extract Medicated Serum

作　　者：付江 刘敬莉[3] 柳科军 段磊 蒲俊 卜阳 FU Jiang;LIU Jingli;LIU Kejun;DUAN Lei;PU Jun;BU Yang(Department of Neurosurgery,Central Hospital of Xiaogan,Xiaogan 432000,China;School of Clinical Medicine,Ningxia Medical University,Yinchuan 750004,China;Department of Hepatobiliary Surgery,General Hospital of Ningxia Medical University,Yinchuan 750004,China;Department of Hepatobiliary Surgery,People's Hospital of Ningxia Hui Autonomous Region,Yinchuan 750002,China)

机构地区：[1]湖北省孝感市中心医院神经外科,孝感432000 [2]宁夏医科大学临床医学院,银川750004 [3]宁夏医科大学总医院肝胆外科,银川750004 [4]宁夏回族自治区人民医院肝胆外科,银川750002

出　　处：《宁夏医科大学学报》2021年第7期665-670,733,共7页Journal of Ningxia Medical University

基　　金：宁夏自然科学基金(NZ15130)。

摘　　要：目的研究银杏叶提取物(ginkgo biloba extract,GBE)含药血清中血小板活化因子(platelet activating factor,PAF)含量,以及此含药血清对肝癌细胞株增殖、侵袭与迁移的影响和相关作用机制。方法采用GBE注射液腹腔注射裸鼠BALB/c nu,2周后获取含药血清,酶联免疫吸附实验(enzyme-linked immunosorbent assay,ELISA)检测裸鼠外周含药血清中PAF水平,含药血清处理人肝癌细胞株MHCC97H和Hep3B,CCK8实验检测肝癌细胞的增殖能力、细胞划痕实验和Transwell实验检测肝癌细胞株的迁移和侵袭能力变化;Western blot进一步验证增殖相关标志物PCNA(proliferating cell nuclear antigen)和上皮间质转化(epithelial-mesenchymal transition,EMT)相关标志物E-cadherin、Vimentin、Snail的变化。结果GBE腹腔注射裸鼠血清中PAF含量降低;GBE含药血清处理肝癌细胞后,细胞增殖、侵袭和迁移能力显著降低;Western blot检测肝癌细胞经GBE含药血清处理后PCNA表达下调,细胞增殖能力降低,且E-cadherin升高、而Vimentin、Snail表达降低,呈间质细胞上皮化改变(mesenchymal-epithelial transition,MET)。结论GBE含药血清能够明显抑制肝癌细胞增殖,逆转肝癌细胞EMT,其作用可能与GBE含药血清中PAF含量降低有关。Objective To study the content of platelet activating factor(PAF)in ginkgo biloba extract(GBE)medicated serum.To explore the effect and mechanism of this medicated serum on proliferation and migration of hepatoma cell lines.Methods BALB/c nude mice were injected with GBE intra-peritonelly,and the serum of nude mice were obtained after 2 weeks.ELISA was used to detect the expression quantity of PAF in serum of nude mice.Treatment of human hepatoma cell lines MHCC97H and Hep3B with GBE medicated serum.CCK8 was used to detect the proliferation ability of hepatoma cells.The migration ability was evaluated by wound healing experiment.The Transwell chamber was also used to detect the invasion ability of the tumor cells.The marker of PCNA protein related to tumor proliferation was detected by Western blot.Furthermore,the expressions of Vimentin,E-cadherin and Snail proteins related to EMT were detected by Western blot.Results ELISA showed that the level of PAF was obviously decreased in serum of nude mice after GBE treatment.With the intervention of serum from nude mice after GBE treatment,the ability of MHCC97H cells and Hep3B cells with regard to proliferation,invasion and migration were significantly decreased.In hepatoma cell with intervention of mice serum treated with GBE,the expression level of PCNA was decreased by Western blot,indicating that the proliferative ability of hepatoma cells inhibited.Meanwhile,the E-cadherin expression was increased.But the expression level of Uimentin and Snail were obviously decreased.It was suggested that the hepatoma cells were MET.Conclusion Serum containing GBE showed decreased level of PAF.It might be related to the inhibition of proliferation,invasion and changes of related protein expression of hepatoma cell line by serum containing GBE.This study was an important supplement to the theoretical mechanism of anti-liver cancer by traditional Chinese medicine GBE,which would provide a theoretical basis for further clinical transformation and application.

关 键 词：银杏叶提取物 肝癌 侵袭 转移 血小板活化因子 

分 类 号：R735.7[医药卫生—肿瘤]