清肝化瘀颗粒对H22肝癌腹水瘤小鼠的治疗作用

Effect of Qinggan Huayu granule on H22 liver ascitic tumor mice

目的:研究清肝化瘀颗粒对H22肝癌腹水瘤小鼠的治疗作用。方法:建立H22肝癌腹水瘤小鼠模型,将120只小鼠随机平均分为模型组、肝复乐组(1.35 g/kg)、氟尿嘧啶组(50 mg/kg,i.p)、清肝化瘀颗粒低剂量组(0.67 g/kg)、清肝化瘀中剂量组(1.34 g/kg)、清肝化瘀高剂量组(2.68 g/kg),连续给药10 d,每3日监测体重及腹围,第11天每组随机选择8只小鼠取材,检测腹水量、腹膜渗透性及腹膜组织病理染色等,剩余小鼠观察生存期。使用Westernblot法检测腹膜组织血管内皮生长因子A(VEGFA)及血管内皮细胞生长因子受体2(VEGFR2)表达量。结果:与模型组相比,氟尿嘧啶组、中剂量组、高剂量组小鼠体重增长较慢(P<0.05);与模型组相比,各用药组小鼠腹围增长较慢,第6天,氟尿嘧啶组、中剂量组腹围显著小于模型组(P<0.05);第12天高剂量组腹围显著小于模型组(P<0.05)。此外,与模型组相比,中、高剂量组小鼠腹水量明显减少(P<0.05);中、高剂量组、氟尿嘧啶组小鼠腹水上清吸光度明显降低(P<0.05);中剂量组和氟尿嘧啶组生存期明显延长(P<0.05)。低剂量组、肝复乐组上述指标的差异无统计学意义。腹膜病理染色显示,氟尿嘧啶组、中、高剂量组小鼠腹膜结构较完整、水肿较轻、血管新生较少,优于肝复乐组和低剂量组。与模型组相比,清肝化瘀中剂量组VEGFA及VEGFR2表达量显著降低(P<0.05,P<0.01)。结论:清肝化瘀颗粒可抑制H22肝癌腹水瘤小鼠腹水产生、延长小鼠生存期,并降低腹膜渗透性、抑制腹膜新生血管的增加,其机制可能与抑制VEGF/VEGFR通路相关。

Objective:To evaluate the therapeutic effect of Qinggan Huayu granule on mice with H22 liver cancer ascites tumor.Methods:A H22 liver cancer ascites mouse model was established by intraperitoneally injecting H22 liver cancer cells.Mice were randomly divided into the model group,the Ganfule group(1.35 g/kg),the fluorouracil group(50 mg/kg i.p),the Qinggan Huayu granule groups at low(0.67 g/kg),medium(1.34 g/kg),and high(2.68 g/kg)doses.Then the mice were ad⁃ministered continuously for 10 d and body weight and abdominal circumference were monitored every 3 d.On day 11,eight rats in each group were randomly selected for dissection to detect the amount of peritoneal water,peritoneal permeability and histopatho logical changes.The remaining mice were observed for survival.In addition,the vascular endothelial growth factor A(VEGFA)and vascular endothelial growth factor receptor 2(VEGFR2)were determined by Western blotting.Results:Compared with the model group,the weight growth of mice in the fluorouracil group and the medium-dose and high-dose Qinggan Huayu granule groups was slower(P<0.05).Moreover,the abdominal circumference of mice in each treatment group was increased slowly.There were significant differences in abdominal circumference between the fluorouracil group,the medium-dose group and the con⁃trol group from day 6(P<0.05)while the abdominal circumference of the high dose group was significantly smaller than that of the control group from day 12(P<0.05).Moreover,compared with the model group,the amount of ascites in the medium-and high-dose Qinggan Huayu granule groups was decreased significantly(P<0.05).The optical density value of ascites supernatant in medium-and high-dose Qinggan Huayu granule group and the fluorouracil group was decreased significantly(P<0.05)and the survival period of the medium-dose Qinggan Huayu granule group and the fluorouracil group was prolonged prominently(P<0.05).There was no significant difference in the low-dose Qinggan Huayu granule group and the Ganfule group.Peritoneal histo⁃pathological assay showed more complete peritoneal structure,less edema and less angiogenesis of the peritoneum in the fluoroura⁃cil group and the medium-and high-dose Qinggan Huayu granule groups,which was better than those of the Ganfule group and the low-dose group.Compared with the model group,the expressions of VEGFA and VEGFR2 in the medium-dose Qinggan Huayu granule group were decreased significantly(P<0.05,P<0.01).Conclusion:Qinggan Huayu granule can inhibit ascites produc⁃tion in the mice model with H22 liver cancer ascites tumor,prolong the survival of mice,reduce peritoneal permeability and sup⁃press the increase of peritoneal neovascularization.The mechanism may be related to the inhibition of VEGF/VEGFR pathway.