抗IL-9 抗体治疗肝癌恶性腹水的疗效及其机制研究

目的:探索抗IL-9 抗体治疗小鼠肝癌恶性腹水(MA)的疗效及作用机制。方法:应用小鼠H22 细胞系建立小鼠MA 模型,造模24 h 后将45 只小鼠随机分成实验组、阴性对照组和空白对照组,每组15 只。实验组给予腹腔注射抗IL-9抗体,阴性对照组给予注射同型IgG 抗体,空白对照组给予注射生理盐水,隔天1 次,共注射5 次。每次注射前均称量各组小鼠体重,并观察小鼠日常活动状态。末次用药结束24 h 后每组随机处死5 只小鼠,测量MA 体积,收集MA 上清,采用ELISA法检测MA 中VEGF、MMP-2、IL-9、IFN-β表达水平。其余小鼠观察其生存时间。结果:干预后实验组、阴性对照组和空白对照组小鼠的MA 体积分别为(6.70±1.52)ml、(10.28±1.75)ml、(10.36±2.30)ml,与空白对照组和阴性对照组相比,实验组小鼠MA 体积明显减少,差异有统计学意义(P<0.05)。实验组小鼠平均生存时间为(17.2±2.1)d,显著长于阴性对照组(14.5±1.2)d 和空白对照组(14.8±1.4)d(P<0.05);与空白对照组和阴性对照组相比,实验组小鼠MA 中VEGF 水平显著降低,差异有统计学意义(P<0.05),与阴性对照组相比,实验组小鼠MA 中IL-9 水平显著降低,差异有统计学意义(P<0.05);但三组间MMP-2、IFN-β水平比较差异无统计学意义(P>0.05)。结论:腹腔注射抗IL-9 抗体可以有效地抑制H22 细胞荷瘤小鼠MA 的产生,延长其生存时间,该作用可能是通过抑制MA 中VEGF、IL-9 的表达而实现。

Effects and mechanism of anti IL-9 antibody on malignant ascites of hepatic carcinoma

Objective:To explore the effects and mechanism of anti IL-9 antibody on malignant ascites (MA) of hepatic carcinoma in mice.Methods: A mouse model of MA was established by mouse H22 cell line.45 mice were divided randomly into experimental group,negative control group and blank control group at 24 hours after modeling,with 15 mice in each group.The experimental group was injected intraperitoneally with anti IL-9 antibody;the negative control group was injected with isotype IgG antibody;the blank control group was injected with normal saline.The weight and behavior of the mice were measured before each injection.Five mice of each group was sacrificed at 24 hours after the last injection to measure the volume of MA.The level of VEGF,MMP-2,IL-9 and IFN-in MA were determined with ELISA assay;the survival time of rest mice were recorded and compared.Results: The mean volume of MA of experimental group,negative control group and blank control group were (6.70±1.52)ml,(10.28±1.5)ml,(10.36±2.30) ml,respectively, the MA volume of experimental group were lower as compared to negative control group and blank control group(P<0.05).The mean survival time of experimental group was (17.2±2.1)d,which was significantly prolonged compared with the negative control group (14.5±1.2)d and the blank control group (14.8±1.4)d (P<0.05).The levels of VEGF of MA in experimental group was significantly lower compared to the negative control group and blank control group (P<0.05).The levels of IL-9 of MA in experimental group was significantly lower compared to the negative control group(P<0.05).The levels of MMP-2 and IFN-βin experimental group had no significant difference compared with the negative control group and blank control group (P>0.05). Conclusion: Intraperitoneal injection anti IL-9 antibody on H22 ascites-bearing mice can effectively inhibit the generation of the MA.The mechanism may be related to the inhibition of the expression of the VEGF and IL-9.