摘要
本研究为探究七叶皂苷钠(aescin sodium, AS)对HepG2肝癌细胞的氧化损伤作用及对Nude无胸腺裸鼠体内成瘤的调控作用及其作用机制。体内试验以Nude无胸腺裸鼠为载体,皮下接种5 106个人肝癌细胞株HepG2建立肿瘤模型。低、中和高剂量七叶皂苷钠组分别腹腔注射1 mg/kg、2.5 mg/kg及5 mg/kg的七叶皂苷钠进行治疗。体外以HepG2细胞为载体,通过CCK8试验检测七叶皂苷钠对HepG2细胞的细胞活力的影响,通过乳酸脱氢酶(LDH)试剂盒检测七叶皂苷钠对HepG2细胞的LDH释放的影响,通过标准试剂盒检测HepG2细胞内氧化损伤因子谷胱甘肽过氧化物酶(GSH-px)、超氧化物歧化酶(SOD)、活性氧(ROS)水平及丙二醛(MAD)的表达,通过荧光定量PCR(real-time PCR)试验检测小鼠肝脏及HepG2细胞内PI3K/Akt/mTOR通路基因表达。结果与Model组相比,七叶皂苷钠处理组显著降低小鼠体内瘤重及增加Nude无胸腺裸鼠体重,且2.5 mg/kg的七叶皂苷钠治疗组有最好的治疗效果。与Control组相比,七叶皂苷钠呈浓度依赖性抑制HepG2细胞的细胞活力,增加细胞内LDH释放及细胞的氧化损伤。且七叶皂苷钠在体内外下调了肝癌中PI3K/Akt/mTOR通路基因的表达。而在使用PI3K/Akt/mTOR通路的激活剂450 Y-P后,七叶皂苷钠的抗肝癌作用被显著减弱。七叶皂苷钠能诱导HepG2肝癌细胞的氧化损伤,抑制Nude无胸腺裸鼠的体内成瘤,并且其可能是通过调控机体内的PI3K/AKT/mTOR通路来实现的。表明七叶皂苷钠对原发性肝癌有一定的抑制作用,具有成为原发性肝癌治疗药物的潜力。
To investigate the anti-cancer effect of aescin sodium(AS) in HepG2 cells and in Nude athymic nude mice. Nude athymic nude mice were used as the in vivo carrier through subcutaneously inoculate 5 106 HepG2 cells to establish a tumor model. 1 mg/kg, 2.5 mg/kg,and 5 mg/kg AS were treated by intraperitoneal injection to cure the tumor model. HepG2 cells were used as an in vitro carrier. The cell viability of HepG2 cells was detected by CCK8 assay, and the release of lactate dehydrogenase(LDH) from HepG2 cells was detected by standard LDH kit. The oxidative damage factors glutathione peroxidase(GSH-px), superoxide dismutase(SOD), reactive oxygen species(ROS)and malondialdehyde(MAD) in HepG2 cells were detected by standard kit. Real-time PCR was used to detect the gene expression of PI3K/Akt/mTOR pathway in mouse liver and HepG2 cells. Results compared with the Model group, AS treatment significantly reduced the tumor weight in mice and increased the body weight of Nude athymic nude mice, and the 2.5 mg/kg AS treatment had the best therapeutic effect.Compared with the control group, AS treatment inhibited the cell viability of HepG2 cells in a concentration-dependent manner, increased the release of intracellular LDH and the oxidative damage of the cells. And AS reduced the gene expression of PI3K/Akt/mTOR pathway in vitro and in vivo. However, the anti-cancer effect of AS was significantly attenuated after the use of 450 Y-P, an activator of the PI3K/Akt/mTOR pathway. AS treatment induced oxidative damage in HepG2 cells and inhibit tumorigenesis in Nude athymic nude mice, which was regulated by the PI3K/AKT/mTOR pathway. It showed that AS has a certain inhibitory effect on HCC and has the potential to become a therapeutic drug for HCC.
作者
刘东生
高成业
张宝书
翟菊敏
李广鑫
LIU Dong-sheng;GAO Cheng-ye;ZHANG Bao-shu;ZHAI Ju-min;LI Guang-xin(The Fifth Department of General Surgery,The First Hospital of Handan City,Handan 056002,China)
出处
《特产研究》
2022年第4期27-33,共7页
Special Wild Economic Animal and Plant Research
基金
邯郸市科学技术研究与发展计划(1523108078-5)。
作者简介
刘东生(1982-),男,河北省邯郸人,硕士,主治医师,从事普外科临床研究。
