Neuropilin1单克隆抗体对胶质瘤大鼠Bcl-2/Bax表达的影响

Effect of Neuropilin1 monoclonal antibody on Bcl-2/Bax expression in glioma rats

目的:研究Neuropilin1单克隆抗体对胶质瘤大鼠Bcl-2/Bax表达的影响。方法:将36只SD大鼠随机分为假手术组、模型组和治疗组,每组12只;假手术组只行颅骨开窗手术,其余各组均建立胶质细胞瘤模型。模型组腹腔注射0.9%氯化钠,治疗组腹腔注射NRP-1 MAb,均每日1次。干预14天后取材。对肿瘤组织称重,免疫组化检测Bax和Bcl-2表达,Western blot检测NRP-1蛋白、Bax蛋白和Bcl-2蛋白表达量,qPCR检测Bax mRNA和Bcl-2 mRNA表达情况,TUNEL检测细胞凋亡情况。结果:假手术组无肿瘤,治疗组肿瘤组织重量显著小于模型组,差异具有统计学意义(P<0.05)。免疫组化显示:与假手术组比较,模型组和治疗组Bax表达显著下降,Bcl-2表达显著上升,差异具有统计学意义(P<0.05);与模型组比较,治疗组Bax表达上升,Bcl-2表达下降,差异具有统计学意义(P<0.05)。Western blot检测显示:与假手术组比较,模型组和治疗组NRP-1蛋白和Bcl-2蛋白表达量显著上升,Bax蛋白表达量显著下降,差异具有统计学意义(P<0.05);与模型组比较,治疗组NRP-1蛋白和Bcl-2蛋白表达量显著下降,Bax蛋白表达量显著上升,差异具有统计学意义(P<0.05)。qPCR检测显示:与假手术组比较,模型组和治疗组Bax mRNA表达显著下降,Bcl-2mRNA表达显著上升,差异具有统计学意义(P<0.05);与模型组比较,治疗组Bax mRNA表达上升,Bcl-2mRNA表达下降,差异具有统计学意义(P<0.05)。TUNEL检测显示:与假手术组比较,模型组和治疗组的细胞凋亡率显著上升,差异具有统计学意义(P<0.05);与模型组比较,治疗组的细胞凋亡率显著下降,差异具有统计学意义(P<0.05)。结论:NRP-1MAb可通过抑制NRP-1表达而上调Bax蛋白表达,下调Bcl-2蛋白表达促进胶质细胞瘤凋亡。

Objective:To study the effect of Neuropilin1 monoclonal antibody on the expression of Bcl-2/Bax in glioma rats.Methods:36 SD rats were randomly divided into sham operation group,model group and treatment group with 12 rats in each group.Sham operation group only underwent craniotomy,and other groups established glioma model.After 10 days,the model group was given 0.9% sodium chloride intraperitoneal injection once a day,while the treatment group was given NRP-1 MAb intraperitoneal injection once a day.The expression of Bax and Bcl-2 was detected by immunohistochemistry.The expression of NRP-1 protein,Bax protein and Bcl-2 protein was detected by Western blot.The expression of Bax mRNA and Bcl-2 mRNA was detected by qPCR,and apoptosis was detected by TUNEL.Results:There was no tumor in sham operation group,and the weight of tumor tissue in treatment group was significantly less than that in model group ( P <0.05).Compared with sham operation group,the expression of Bax in model group and treatment group was decreased significantly ( P <0.05),and the expression of Bcl-2 was increased significantly ( P <0.05).Compared with the model group,the expression of Bax in the treatment group was increased,and the expression of Bcl-2 was decreased ( P <0.05).Western blot analysis showed that the expression of NRP-1 protein and Bcl-2 protein in the model group and the treatment group was increased significantly,while the expression of Bax protein was decreased significantly ( P <0.05).Compared with the model group,the expression of NRP-1 protein and Bcl-2 protein in the treatment group was decreased significantly,while the expression of Bax protein was increased significantly ( P <0.05).The difference was statistically significant ( P <0.05).Compared with the sham operation group,the expression of Bax mRNA in the model group and the treatment group was significantly decreased,and the expression of Bcl-2 mRNA was significantly increased ( P <0.05).Compared with the model group,Bax mRNA expression of the treatment group was incresased and Bcl-2 mRNA expression was decreased ( P <0.05).TUNEL test showed that compared with sham operation group,the apoptosis rates of the model group and the treatment group were increased significantly ( P <0.05).Compared with the model group,the apoptosis rate of the treatment group was decreased significantly ( P <0.05).Conclusion:NRP-1 MAb can promote the apoptosis of glioma by inhibiting the expression of NRP-1 and up-regulating the expression of Bax protein and down-regulating the expression of Bcl-2 protein.