有氧运动联合枸杞多糖在抑制小鼠H_(22)肝肿瘤过程中mTOR作用的研究

Effect of aerobic exercise and Lycium Barbarum Polysaccharide on expression of mTOR in H_(22) tumor-bearing mice

目的探讨有氧运动联合枸杞多糖对小鼠H_(22)肝肿瘤模型雷帕霉素靶蛋白(mTOR)信号通路的抑制作用,以及抑制mTOR信号通路对H_(22)肝肿瘤增殖的影响。方法 40只雄性昆明种小鼠,构模成功小鼠随机分为模型组(C组)、枸杞多糖组(L组)、有氧运动组(E组)和有氧运动联合枸杞多糖组(EL组)4组,每组10只。观察肝肿瘤体积,计算抑瘤率,HE染色观察,蛋白质印迹法检测肿瘤AMPK、mTOR和Survivin蛋白。结果各组荷瘤小鼠皮下肿瘤抑制率,L组为36.09%,E组为20.19%,EL组为54.55%。HE染色发现,C组肿瘤排列较密集、规则,E组肿瘤排列欠规则并出现较小坏死区,L组肿瘤出现坏死区且比E组明显,EL组肿瘤内部可见明显坏死区。蛋白质印迹法检测提示,L组、E组和EL组肿瘤组织AMPK蛋白表达量分别为1.54±0.23、1.09±0.17和2.03±0.29,均较C组增加,均P<0.001。与C组、L组、E组比较,EL组AMPK蛋白表达最高,均P<0.001。L组、E组和EL组肿瘤组织mTOR蛋白表达量分别为1.98±0.23、2.41±0.29和1.53±0.17,均较C组降低,F=25.876,P<0.001。与C组、L组、E组比较,EL组mTOR蛋白表达最低,均P<0.001。L组、E组和EL组肿瘤组织Survivin蛋白表达量分别为1.64±0.14、2.06±0.23和1.25±0.25,均较C组降低,F=15.457,P<0.001。与C组、L组、E组比较,EL组Survivin蛋白表达最低,均P<0.001。结论有氧运动联合枸杞多糖降通过下调H_(22)肝肿瘤细胞mTOR蛋白的表达抑制小鼠H_(22)肝肿瘤细胞增殖。

Objective To demonstrate the effect of aerobic exercise and Lycium Barbarum Polysaccharide on the inactivation of mTOR pathway in mice,and investigated inactivation of this signaling pathway inhibited tumor proliferation.Methods Totally40 KM mice,H_(22) tumor-bearing mice were randomly divided into 4 groups:control group(C),Lycium Barbarum Polysaccharide group(L),aerobic exercise group(E),aerobic exercise and Lycium Barbarum Polysaccharide(EL),each group of10 mice.The tumor volumes were measured.The tumor inhibition ratios were calculated,and the tumor cell was observed with HE staining.AMPK,mTOR,and Survivin protein expression were detected by Western Blotting.Results The tumor inhibitory rates of mice were 36.09%in L group,20.19%in E group and 54.55%in EL group.The tumor inhibitory rate of mice in EL group was significantly higher than that in other groups.Morphology of tumor cells in the C group appeared to be arranged densely.E group cells arranged in irregular with small necrosis area and obvious necrosis in L group,large necrotic area in EL group.AMPK expression showed a significant increase than that in the control group(P<0.001)by Western Blotting,which were 1.54±0.23,1.09±0.17 land 2.03±0.29 in L group,E group and EL group respectively.Compared with C group,L group and E group,AMPK protein in EL group was the highest,all P<0.001.mTOR expression showed a significant decrease than that in the control group(F=25.876,P<0.001)by Western Blotting,which were 1.98±0.23,2.41±0.29,1.53±0.17 in L group,E group and EL group respectively.mTOR protein in EL group was the lowest compared with C group,L group and E group,all P<0.001.Similarly,Survivin protein showed a significant decrease than that in the control group(F=15.457,P<0.001)by Western Blotting,which were1.64±0.14,2.06±0.23,1.25±0.25 in L group,E group and EL group respectively.Compared with C group,L group and E group,Survivin protein in EL group was the lowest,all P<0.001.Conclusion Aerobic exercise and Lycium barbarum polysaccharide inhibited the proliferation of H_(22) hepatoma cells in mice by down-regulating the expression of mTOR protein.