VEGF／bFGF 复合多肽疫苗对雌性小鼠的毒性及其生殖的影响

摘要

目的：初步探究血管内皮生长因子（ VEGF ）／碱性成纤维细胞生长因子（ bFGF ）复合多肽疫苗（ VEGF／bFGF complex peptide vaccine，VBP3）对雌性小鼠的毒性及其生殖的影响。方法：通过镍离子亲和层析柱纯化VBP3蛋白，将纯化获得的VBP3免疫雌性BALB／c小鼠，酶联免疫吸附实验（ ELISA）方法检测小鼠血清抗体效价和特异性，监测亲本小鼠体重，测定亲本小鼠脏器重量并对小鼠脏器进行HE染色观察；将免疫小鼠与未免疫雄性小鼠交配后，测定F1代小鼠存活率、体重、相关脏器重量，并对相关脏器进行HE染色观察。结果：间接ELISA检测结果显示免疫小鼠血清中抗VEGF、抗bFGF抗体滴度分别为1∶3000、1∶20000，且免疫组F1代小鼠能检测出低滴度的抗bFGF抗体，但抗VEGF抗体不能检出。在小鼠生产率实验中，免疫组与对照组崽鼠数目未见明显差异，但免疫组F1代小鼠存活率较对照组降低（ P＜0．05）。在亲本小鼠中，免疫组小鼠各脏器重量与对照组比较均未有差异，而在F1代小鼠中，2个组别的小鼠肝脏重量存在差异，但其它脏器重量未有差异；HE染色显示，在亲本小鼠中，2个组别小鼠的各脏器形态未有明显差异；在F1代小鼠中，免疫组F1代小鼠的肝脏与对照组相比存在形态差异。结论：VEGF／bFGF复合多肽疫苗未对亲本小鼠主要脏器造成直接损伤，但对亲本小鼠的生殖及F1代小鼠健康具有一定的影响。%[ ABSTRACT ] AIM: To investigate the toxicity and reproductive effect of vascular endothelial growth factor ( VEGF)/basic fibroblast growth factor ( bFGF) complex peptide vaccine ( VBP3) on the female-mice.METHODS:The VBP3 was purified with Ni-NTA affinity chromatography.The female BALB/c mice were immunized with the purified VBP3.The antibody titer in the serum was detected by ELISA.The data of the body weight and the organ weight of the par-ent mice were gathered and analyzed, and the pathological changes of the organs were observed with HE staining to investi-gate the toxicity of VBP3.To investigate the toxicity of VBP3 in the F1 mice, the parent immunized female mice were ma-ted with the parent non-immunized male mice.After the F1 mice were born, the survival rate was calculated, the change of the body weight and the organ weight of the F1 mice were also determined.The pathological changes of the organs in F1 mice were also observed with HE staining.RESULTS: In the parent mice, high titers of the antibodies were detected against VEGF and bFGF, and no difference of the body weight, the organ weight and the pathological change between the immunized mice and control mice was found.In the F1 mice, a low titer of anti-bFGF antibody was detected compared with blank group.The survival rate in control group was higher than that in immunized group.In the 2 groups of the F1 mice, no obvious difference of the weight of the spleen, kidney, lung and heart was observed, and there was some difference in the weight of liver between the 2 groups.The results of the HE staining in the F1 mice showed some difference in the liver between the 2 groups.CONCLUSION:VEGF/bFGF complex peptide vaccine has no obvious organ toxicity in the parent female mice, but has some side effects on the reproductive and the healthy processes of F1 mice.