AIM To investigate the antitumor activity of α-hederin in hepatocellular carcinoma(HCC) cells and its underlying mechanisms in vitro and in vivo.METHODS SMMC-7721, Hep G-2 and Huh-7 HCC cells were cultured in vitro a...AIM To investigate the antitumor activity of α-hederin in hepatocellular carcinoma(HCC) cells and its underlying mechanisms in vitro and in vivo.METHODS SMMC-7721, Hep G-2 and Huh-7 HCC cells were cultured in vitro and treated with α-hederin(0, 5 μmol/L, 10 μmol/L, 15 μmol/L, 20 μmol/L, 25 μmol/L, 30 μmol/L, 35 μmol/L, 40 μmol/L, 45 μmol/L, 50 μmol/L, 55 μmol/L, or 60 μmol/L) for 12 h, 24 h, or 36 h, and cell viability was then detected by the Cell Counting Kit-8. SMMC-7721cells were treated with 0, 5 μmol/L, 10 μmol/L, or 20 μmol/L α-hederin for 24 h with or without DL-buthionineS,R-sulfoximine(2 mmol/L) or N-acetylcysteine(5 mmol/L) pretreatment for 2 h, and additional assays were subsequently performed. Apoptosis was observed after Hoechst staining. Glutathione(GSH) and adenosine triphosphate(ATP) levels were measured using GSH and ATP Assay Kits. Intracellular reactive oxygen species(ROS) levels were determined by measuring the oxidative conversion of 2',7'-dichlorofluorescin diacetate. Disruption of the mitochondrial membrane potential was evaluated using JC-1 staining. The protein levels of Bax, Bcl-2, cleaved caspase-3, cleaved caspase-9, apoptosis-inducing factor and cytochrome C were detected by western blotting. The antitumor efficacy of α-hederin in vivo was evaluated in a xenograft tumor model.RESULTS The α-hederin treatment induced apoptosis of HCC cells. The apoptosis rates in the control, low-dose α-hederin(5 μmol/L), mid-dose α-hederin(10 μmol/L) and highdose α-hederin(20 μmol/L) groups were 0.90% ± 0.26%, 12% ± 2.0%, 21% ± 2.1% and 37% ± 3.8%, respectively(P < 0.05). The α-hederin treatment reduced intracellular GSH and ATP levels, induced ROS, disrupted the mitochondrial membrane potential, increased the protein levels of Bax, cleaved caspase-3, cleaved caspase-9, apoptosis-inducing factor and cytochrome C, and decreased Bcl-2 expression. The α-hederin treatment also inhibited xenograft tumor growth in vivo. CONCLUSION The α-hederin saponin induces apoptosis of HCC cells via the mitochondrial pathway mediated by increased intracellular ROS and may be an effective treatment for human HCC.展开更多
OBJECTIVEα-Hederin is an effective component of the traditional Chinese medicine Pulsatilla chinensis,which has been reported to exert many pharmacological activities.However,the effect ofα-hederin on metabolism is ...OBJECTIVEα-Hederin is an effective component of the traditional Chinese medicine Pulsatilla chinensis,which has been reported to exert many pharmacological activities.However,the effect ofα-hederin on metabolism is still unclear.This study aimed to illuminate the role ofα-hederin in glucose metabolism in lung cancer cells and investigate the molecular mechanism ofα-hederin.METHODS CCK8 and colony formation assays were employed to assess the anti-proliferative effects induced byα-hederin.Glucose uptake,ATP generation,and reduced lactate production were measured using kits,and an A549 tumor xenograft mouse model of lung cancer was used to assess the in vivo antitumor effect ofα-hederin(5,10 mg·kg^-1).Glycolytic-related key enzymes were detected by Western blotting and immunohisto⁃chemical staining.RESULTS Cell proliferation was significantly inhibited byα-hederin in a dose-dependent manner and thatα-hederin inhibited glucose uptake and ATP generation and reduced lactate production.Furthermore,α-hederin remarkably inhibited hexokinase 2(HK2),glucose transporters 1(GLUT1),pyruvate kinase M2(PKM2),lactate dehydro⁃genase A(LDHA),monocarboxylate transporter(MCT4),c-Myc,and hypoxia inducible factor-1α(HIF-1α)protein expres⁃sion.Using inhibitors,we proved thatα-hederin inhibits glycolysis by inhibiting glycolytic regulators.Moreover,a tumor xenograft mouse model of lung cancer further confirmed thatα-hederin inhibits lung cancer growth via inhibiting glycolysis in vivo.CONCLUSIONα-Hederin inhibits the growth of non-small cell lung cancer A549 cells by inhibiting glycolysis.The mechanism of glycolysis inhibition includesα-hederin inhibiting the expression of the glycolytic regulatory factors HIF-1α and c-Myc.展开更多
[目的]建立黄褐毛忍冬(Lonicera fulvotomentosa Hsu et S.C.Cheng)总皂苷的水提工艺。[方法]以黄褐毛忍冬总皂苷中有效成分α-常春藤皂苷的含量为考察指标,采用正交设计法考察加水量、煎煮时间、煎煮次数及浸泡时间4个因素对黄褐毛忍...[目的]建立黄褐毛忍冬(Lonicera fulvotomentosa Hsu et S.C.Cheng)总皂苷的水提工艺。[方法]以黄褐毛忍冬总皂苷中有效成分α-常春藤皂苷的含量为考察指标,采用正交设计法考察加水量、煎煮时间、煎煮次数及浸泡时间4个因素对黄褐毛忍冬总皂苷提取工艺的影响,同时采用高效液相法测定α-常春藤皂苷的含量。[结果]加水量、煎煮次数、煎煮时间、浸泡时间对α-常春藤皂苷含量的影响程度不同,各因素影响程度的大小为:煎煮次数>加水量>浸泡时间>煎煮时间,即煎煮次数对煎煮条件影响最大,为最重要的因素,其次为加水量、浸泡时间、煎煮时间。最优方案为:黄褐毛忍冬药材不用浸泡,加10倍量水煎煮3次,每次1.0h。方差分析结果表明,加水量和煎煮次数对α-常春藤皂苷含量的影响具有显著意义(PA=0.034<0.05,PB=0.020<0.05),而煎煮时间、浸泡时间对水α-常春藤皂苷含量无显著影响(PC=0.500>0.05,PD=0.264>0.05),对测定结果影响小,这与直观分析结果相吻合。[结论]优选提取工艺简单、稳定、可行。展开更多
基金Supported by the National Natural Science Foundation of China,No.81572426the Natural Science Foundation of Hubei Province,No.2015CKB755
文摘AIM To investigate the antitumor activity of α-hederin in hepatocellular carcinoma(HCC) cells and its underlying mechanisms in vitro and in vivo.METHODS SMMC-7721, Hep G-2 and Huh-7 HCC cells were cultured in vitro and treated with α-hederin(0, 5 μmol/L, 10 μmol/L, 15 μmol/L, 20 μmol/L, 25 μmol/L, 30 μmol/L, 35 μmol/L, 40 μmol/L, 45 μmol/L, 50 μmol/L, 55 μmol/L, or 60 μmol/L) for 12 h, 24 h, or 36 h, and cell viability was then detected by the Cell Counting Kit-8. SMMC-7721cells were treated with 0, 5 μmol/L, 10 μmol/L, or 20 μmol/L α-hederin for 24 h with or without DL-buthionineS,R-sulfoximine(2 mmol/L) or N-acetylcysteine(5 mmol/L) pretreatment for 2 h, and additional assays were subsequently performed. Apoptosis was observed after Hoechst staining. Glutathione(GSH) and adenosine triphosphate(ATP) levels were measured using GSH and ATP Assay Kits. Intracellular reactive oxygen species(ROS) levels were determined by measuring the oxidative conversion of 2',7'-dichlorofluorescin diacetate. Disruption of the mitochondrial membrane potential was evaluated using JC-1 staining. The protein levels of Bax, Bcl-2, cleaved caspase-3, cleaved caspase-9, apoptosis-inducing factor and cytochrome C were detected by western blotting. The antitumor efficacy of α-hederin in vivo was evaluated in a xenograft tumor model.RESULTS The α-hederin treatment induced apoptosis of HCC cells. The apoptosis rates in the control, low-dose α-hederin(5 μmol/L), mid-dose α-hederin(10 μmol/L) and highdose α-hederin(20 μmol/L) groups were 0.90% ± 0.26%, 12% ± 2.0%, 21% ± 2.1% and 37% ± 3.8%, respectively(P < 0.05). The α-hederin treatment reduced intracellular GSH and ATP levels, induced ROS, disrupted the mitochondrial membrane potential, increased the protein levels of Bax, cleaved caspase-3, cleaved caspase-9, apoptosis-inducing factor and cytochrome C, and decreased Bcl-2 expression. The α-hederin treatment also inhibited xenograft tumor growth in vivo. CONCLUSION The α-hederin saponin induces apoptosis of HCC cells via the mitochondrial pathway mediated by increased intracellular ROS and may be an effective treatment for human HCC.
基金National Natural Science Foundation of China(8146061881860720+1 种基金81660683)Scientific Foundation of Double World-classes Subject Development of Jiangxi University of TCM(JXSYLXK-ZHYAO124)
文摘OBJECTIVEα-Hederin is an effective component of the traditional Chinese medicine Pulsatilla chinensis,which has been reported to exert many pharmacological activities.However,the effect ofα-hederin on metabolism is still unclear.This study aimed to illuminate the role ofα-hederin in glucose metabolism in lung cancer cells and investigate the molecular mechanism ofα-hederin.METHODS CCK8 and colony formation assays were employed to assess the anti-proliferative effects induced byα-hederin.Glucose uptake,ATP generation,and reduced lactate production were measured using kits,and an A549 tumor xenograft mouse model of lung cancer was used to assess the in vivo antitumor effect ofα-hederin(5,10 mg·kg^-1).Glycolytic-related key enzymes were detected by Western blotting and immunohisto⁃chemical staining.RESULTS Cell proliferation was significantly inhibited byα-hederin in a dose-dependent manner and thatα-hederin inhibited glucose uptake and ATP generation and reduced lactate production.Furthermore,α-hederin remarkably inhibited hexokinase 2(HK2),glucose transporters 1(GLUT1),pyruvate kinase M2(PKM2),lactate dehydro⁃genase A(LDHA),monocarboxylate transporter(MCT4),c-Myc,and hypoxia inducible factor-1α(HIF-1α)protein expres⁃sion.Using inhibitors,we proved thatα-hederin inhibits glycolysis by inhibiting glycolytic regulators.Moreover,a tumor xenograft mouse model of lung cancer further confirmed thatα-hederin inhibits lung cancer growth via inhibiting glycolysis in vivo.CONCLUSIONα-Hederin inhibits the growth of non-small cell lung cancer A549 cells by inhibiting glycolysis.The mechanism of glycolysis inhibition includesα-hederin inhibiting the expression of the glycolytic regulatory factors HIF-1α and c-Myc.
文摘[目的]建立黄褐毛忍冬(Lonicera fulvotomentosa Hsu et S.C.Cheng)总皂苷的水提工艺。[方法]以黄褐毛忍冬总皂苷中有效成分α-常春藤皂苷的含量为考察指标,采用正交设计法考察加水量、煎煮时间、煎煮次数及浸泡时间4个因素对黄褐毛忍冬总皂苷提取工艺的影响,同时采用高效液相法测定α-常春藤皂苷的含量。[结果]加水量、煎煮次数、煎煮时间、浸泡时间对α-常春藤皂苷含量的影响程度不同,各因素影响程度的大小为:煎煮次数>加水量>浸泡时间>煎煮时间,即煎煮次数对煎煮条件影响最大,为最重要的因素,其次为加水量、浸泡时间、煎煮时间。最优方案为:黄褐毛忍冬药材不用浸泡,加10倍量水煎煮3次,每次1.0h。方差分析结果表明,加水量和煎煮次数对α-常春藤皂苷含量的影响具有显著意义(PA=0.034<0.05,PB=0.020<0.05),而煎煮时间、浸泡时间对水α-常春藤皂苷含量无显著影响(PC=0.500>0.05,PD=0.264>0.05),对测定结果影响小,这与直观分析结果相吻合。[结论]优选提取工艺简单、稳定、可行。
