人参皂苷Rg3联合顺铂抑制小鼠肝细胞癌转移及微血管生成的机制研究

Inhibitory effect and mechanisms of ginsenoside Rg3 combined with cisplatin on the metastasis and microangiogenesis of hepatocellular carcinoma in mice

目的探讨人参皂苷Rg3联合顺铂对小鼠肝细胞癌转移及微血管生成的抑制作用及可能的机制。方法建立KM小鼠H22肝癌模型60只,随机分为6组,根据处理方式的不同分为模型组、顺铂组、Rg3低剂量、Rg3高剂量组、低剂量Rg3联合顺铂组、高剂量Rg3联合顺铂组,检测记录各组小鼠肿瘤质量、肿瘤微血管密度(MVD),采用Western blot法检测瘤基质金属蛋白酶2(MMP2)、基质金属蛋白酶9(MMP9)、血管内皮生长因子(VEGF)蛋白表达。结果(1)治疗14 d后,与模型组比较,各治疗组小鼠体重增长速度均出现不同程度下降。两两比较后发现,高剂量联合组、顺铂组的体重增长幅度最小,Rg3低剂量组及高剂量组的体重增长幅度最为显著(P<0.05)。抑瘤率方面,高剂量联合组的抑瘤率最高,顺铂组及低剂量联合组次之,Rg3低/高剂量组抑瘤率最低(P<0.05)。(2)与模型组比较,各治疗组小鼠血清CD3^+、CD4^+水平均显著升高,CD8^+水平均显著下降(P<0.05)。两两比较后发现,高剂量联合组CD3^+、CD4^+水平最高,低剂量联合组次之,Rg3低/高剂量组低于低剂量联合组(P<0.05)。(3)与模型组比较,各治疗组小鼠肿瘤组织中MMP2、MMP9、VEGF蛋白表达量均出现明显下调(P<0.05)。其中,高剂量联合组相关蛋白表达量最低(P<0.05)。(4)模型组、顺铂组、Rg3低剂量组、Rg3高剂量组、低剂量联合组、高剂量联合组的MVD分别为(12.64±1.96)、(7.73±1.65)、(9.92±1.45)、(7.13±1.64)、(8.16±1.44)、(6.28±1.49),差异具有统计学意义(P<0.05)。结论人参皂苷Rg3可有效抑制H22肝癌细胞的侵袭转移,并与顺铂有一定的协调作用,其机制可能与下调肿瘤转移相关蛋白表达水平、减少肿瘤微血管密度有关。

Objective To investigate the inhibitory effect of ginsenoside Rg3 combined with cisplatin on the metastasis and microangiogenesis of hepatocellular carcinoma in mice and its possible mechanism.Methods Sixty Kunming mice were used to establish H22 hepatocellular carcinoma model and were randomly divided into six groups according to the different treatment method:model group,cisplatin group,low dose Rg3 group,high dose Rg3 group,low dose Rg3 combination group and high dose Rg3 combination group.The microvascular density(MVD),and the expression of matrix metalloproteinase-2(MMP2),matrix metalloproteinase-9(MMP9)and vascular endothelial growth factor(VEGF)in each group was detected.Results (1)The body weight of the high dose Rg3 combination group and cisplatin group was increased by the smallest amount,while that of the Rg3 low dose group and high dose group increased most significantly(P<0.05).(2)Compared with the model group,the serum levels of CD3^+ and CD4^+ cells in each treatment group were increased significantly,while the levels of CD8^+ cells were decreased significantly(P<0.05).The levels of CD3^+ and CD4^+ cells were the highest in the high dose Rg3 group,followed by the low dose Rg3 group,and the CD3^+ and CD4^+ cells in the low/high dose Rg3 group was lower than that in the low dose Rg3 group(P<0.05).(3)Compared with the model group,the expressions of MMP2,MMP9 and VEGF in the tumor tissues of mice in each treatment group were decreased significantly(P<0.05).Among them,the expressions of related proteins in the high dose Rg3 combination group were significantly reduced(P<0.05).(4)The MVD of model group,platinum group,Rg3 low dose group,Rg3 high dose group,low dose combined group and high dose combined group were(12.64±1.96),(7.73±1.65),(9.92±1.45),(7.13±1.64),(8.16±1.44),and(6.28±1.49),respectively.There were significant differences in MVD among the six groups(P<0.05).Conclusions Ginsenoside Rg3 inhibits the invasion and metastasis of H22 hepatocellular carcinoma cells and has a certain coordination with cisplatin.