腹腔内连续或间断应用重组人血管内皮抑素治疗H22腹水瘤的实验研究

Research on continuous or intermittent administration of recombinant human endostatin by intraperitoneal injection in mice H22 ascite tumor model

目的观察重组人血管内皮抑素(恩度)腹腔内连续或间断给药治疗小鼠H22腹水瘤的有效性和安全性,并探索其作用机制。方法利用小鼠H22肝癌细胞系建立腹水瘤动物模型。将102只造模后的ICR小鼠随机分为3组:恩度连续用药组(4mg/kg,12h腹腔注射1次,连续给药8天)、恩度间断用药组(16mg/kg,24h腹腔注射1次,给药4天,休息4天)和对照组(生理盐水,24h腹腔注射1次,连续给药8天),每组34只。记录各组小鼠生存期、腹水体积、体重、腹膜渗透性、腹水中肿瘤细胞数和红细胞数;采用ELISA法检测各组腹水中血管内皮生长因子(VEGF)和基质金属蛋白酶-2(MMP-2)的浓度;检测各组血常规和肝肾功能。结果恩度连续用药组小鼠的平均生存期最长,为(13.60±2.23)d;腹水体积为(8.25±1.45)ml,腹膜渗透性为1.04±0.37,腹水中红细胞数为(0.28±0.06)×108/ml,3项指标均低于恩度间断用药组和对照组(P<0.05);腹水中肿瘤细胞数为(13.7±3.6)×107/ml,低于对照组(P<0.05)。恩度连续用药组的VEGF浓度为(23.64±3.96)pg/ml,MMP-2为(8.94±1.16)ng/ml,显著低于恩度间断用药组和对照组(P<0.05)。恩度连续用药组和恩度间断用药组小鼠的血常规和肝肾功能均未发现明显异常。结论恩度腹腔内应用治疗小鼠H22腹水瘤疗效确切,连续用药的疗效优于间断用药,安全性较好,其机制可能与其抑制VEGF和MMP-2的表达有关。

Objective To observe the effect and safety of recombinant human endostatin（endostar） intraperitoneal injection continuously or intermittently in mice H22 ascite tumor model,and investigate the mechanism.Methods Mice ascite tumor model was established by intraperitoneal injection of H22 cells.One hundred and two ICR mice were randomly assigned into three groups（34 mice in each group）： continuous administration of endostar group（4mg/kg intraperitoneal injection every 12 hours for 8 days）,intermittent administration of endostar group（16mg/kg intraperitoneal injection every 24 hours,administered for 4 days,then have a rest for 4 days）,and control group（intraperitoneal injection normal saline every 24 hours for 8 days）.Weight,volume of ascite,permeability of peritoneal capillaries,number of tumor cells and erythrocytes,lifecycle of mice,blood routine,liver function and renal function in each group were recorded respectively.ELISA method was employed to measure the concentration of VEGF and MMP-2 in ascite.Results In continuous administration of endostar group,the survival time（13.60±2.23）d was the longest among three groups;the volume of ascite was（8.25±1.45）ml,the permeability of peritoneal capillaries was 1.04±0.37,and the number of erythrocyte in the ascite was（0.28±0.06）×108/ml,lower than those in intermittent administration of endostar group and control group（P0.05）.The number of tumor cell in the ascite was（13.7±3.6）×107/ml in continuous administration of endostar group,lower than control group（P0.05）.The concentration of VEGF and MMP-2 in continuous administration of endostar group was（23.64±3.96） pg/ml and（8.94±1.16）ng/ml,lower than those in intermittent administration of endostar group and control group（P0.05）.Blood routine,liver function and renal function of the two endostar groups were normal.Conclusion The intraperitoneal injection of endostar is an effective therapy for mice H22 ascite tumor,and endostar continuously usage is better than intermittently usage.The potential mechanism may be inhibiting the level of VEGF and MMP-2 protein.