藏红花素通过PI3K/Akt/mTOR通路促进人肝癌HepG2细胞凋亡和自噬  

Crocin promotes apoptosis and autophagy of human hepatoma HepG2 cells through PI3K/Akt/mTOR pathway

作　　者：赵彩莎 张运豪 肖婷 ZHAO Cai-sha;ZHANG Yun-hao;XIAO Ting(Seventh Department of Hepatology,Handan Infectious Disease Hospital,Handan 056008;First Department of Spleen and Stomach,Handan Traditional Chinese Medicine Hospital,Handan 056001;First Department of Hepatology,Handan Infectious Disease Hospital,Handan 056008,China)

机构地区：[1]邯郸市传染病医院肝七科,河北邯郸056008 [2]邯郸市中医院脾胃一科,河北邯郸056001 [3]邯郸市传染病医院肝一科,河北邯郸056008

出　　处：《解剖科学进展》2024年第3期287-290,294,共5页Progress of Anatomical Sciences

基　　金：河北省医学科学研究课题(20211577)。

摘　　要：目的研究藏红花素对人肝癌HepG2细胞凋亡和自噬的影响及机制。方法以不同浓度藏红花素(0、0.1、0.2、0.4、0.8、1.6、3.2 mmol/L)干预对数生长期HepG2细胞,48 h后MTT法检测细胞增殖抑制率,计算半抑制浓度(IC50),以IC50作为后续实验药物浓度。设空白对照组、藏红花素组、藏红花素+IGF-1(PI3K激动剂)组、藏红花素+LY294002(PI3K抑制剂)组,药物干预48 h后,MTT法、Annexin V-FITC/PI双染法检测细胞增殖抑制率和凋亡率;GFP-LC3荧光质粒转染HepG2细胞后观察自噬小体;Western blot法检测Cleaved Caspase-3、Cleaved Caspase-9、LC3、Beclin-1、PI3K、p-PI3K、Akt、p-Akt、mTOR、p-mTOR蛋白表达。结果藏红花素对HepG2细胞增殖抑制作用呈现一定的剂量依赖性,IC50值为0.78 mmol/L。与空白对照组比较,藏红花素组HepG2细胞增殖抑制率、凋亡率及自噬小体数量显著升高(P<0.05),Cleaved Caspase-3、Cleaved Caspase-9、LC3-Ⅰ、LC3-Ⅱ、Beclin-1表达量及LC3-Ⅱ/LC3-Ⅰ比值显著升高(P<0.05),PI3K、Akt、mTOR磷酸化水平(p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR)显著降低(P<0.05);IGF-1能够明显逆转藏红花素对HepG2细胞的上述作用,LY294002则能够明显加强藏红花素对HepG2细胞的上述作用(P<0.05)。结论藏红花素具有促进人肝癌HepG2细胞凋亡和自噬的作用,其机制可能与抑制PI3K/Akt/mTOR通路有关。Objective To study the effect and mechanism of Crocin on apoptosis and autophagy of human hepatoma HepG2 cells.Methods Different concentrations of Crocin(0,0.1,0.2,0.4,0.8,1.6,3.2 mmol/L)were used to intervene HepG2 cells in logarithmic growth phase for 48 h,the proliferation inhibition rate was detected by MTT method,and the half-inhibitory concentration(IC50)was calculated.The IC50 was used as the drug concentration in subsequent experiments.The blank control group,Crocin group,Crocin+IGF-1(PI3K activator)group and Crocin+LY294002(PI3K inhibitor)group were set up.Forty-eight hours after drug intervention,the proliferation inhibition rate and the apoptosis rate were respectively detected by MTT or Annexin V-FITC/PI double staining method.The autophagosomes was observated after transfection of GFP-LC3 fluorescent plasmid into HepG2 cells.The expression of Cleaved Caspase-3,Cleaved Caspase-9,LC3,Beclin-1,PI3K,p-PI3K,Akt,p-Akt,mTOR,p-mTOR were detected by Western blot.Results The inhibitory effect of Crocin on HepG2 cell proliferation was dose-dependent,which IC50 value was 0.78 mmol/L.Compared with the blank control group,the proliferation inhibition rate,apoptosis rate and the autophagosomes number of HepG2 cell in Crocin group were significantly increased(P<0.05);the expressions of Cleaved Caspase-3,Cleaved Caspase-9,LC3-Ⅰ,LC3-Ⅱ,Beclin-1 and the ratio of LC3-Ⅱ/LC3-Ⅰwere significantly increased(P<0.05);the phosphorylation level of PI3K,Akt,mTOR(p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR)were significantly decreased(P<0.05).IGF-1 could significantly reverse the above-mentioned effects of Crocin on HepG2 cells,while LY294002 could significantly enhance the above-mentioned effects of Crocin on HepG2 cells,the differences were significant(P<0.05).Conclusion Crocin can promote the apoptosis and autophagy of human hepatoma HepG2 cells,which mechanism may be related to the inhibition of PI3K/Akt/mTOR pathway.

关 键 词：藏红花素 肝癌 PI3K/Akt/mTOR通路 凋亡 自噬 

分 类 号：R735.7[医药卫生—肿瘤]