酶法处理短棒状杆菌的效果分析

Analysis of effect of enzyme-digested Corynebacterium parvum product

目的探讨酶法处理短棒状杆菌(Enzyme-digested Corynebacterium parvum product,ECPP)的效果。方法采用热灭活、超声、Pronase酶消化等方法处理短棒状杆菌(Corynebacterium parvum,CP),电镜观察其结构,并测定其蛋白、多糖及核酸含量。将BALB/c纯系小鼠、青紫蓝家兔和NIH小鼠均随机分为阴性对照组(CK,0.9%NaCl)、短棒状杆菌制剂(C.parvum product,CPP)组(1 000μg/ml CPP)、低剂量ECPP组(ECPPⅠ组,500μg/ml ECPP)和高剂量ECPP组(ECPPⅡ组,1 000μg/mlECPP),观察BALB/c纯系小鼠脾激活情况、异常毒性及血清IL-2、IFNγ及TNF-α的水平,青紫兰家兔热原反应,NIH小鼠抑瘤效果。结果电镜观察可见,与热灭活的CP相比,ECPP细胞壁外围厚度明显变薄,细菌内部结构无明显变化;ECPP的蛋白、多糖及核酸含量较CPP均明显下降,且差异均有统计学意义(P<0.01);ECPPⅡ组小鼠的脾激活指数为2.96,能抑制艾氏腹水瘤,存活率为80%,热原试验合格,异常毒性试验显示其体重增长较CPP组显著,能显著提高小鼠血清中IL-2、IFNγ及TNF-α3种细胞因子的水平。结论酶法处理短棒状杆菌制剂可降低副反应,提高机体免疫功能,具有良好的抗肿瘤作用,是一种有应用前景的新免疫调节剂。

Objective To investigate the effect of enzyme-digested Corynebacterium parvum product（ECCP）.Methods ECPP was prepared by heating inactivation,ultrasonication and Pronase digestion,observed for structure by electron microscopy,and determined for protein,polysaccharide and nucleic acid contents.Pure BALB / c mice,Chinchilla rabbits and NIH mice were randomly divided into negative control（CK,0.9% sodium chloride）,C.parvum product（CPP,1 000 μg / ml） as well as ECPPⅠ（ECPP 500 μg / ml） and Ⅱ（ECPP 1 000 μg / ml） groups separately.The pure BALB / c mice were determined for spleen index,abnormal toxicity and serum IL-2,IFNγ and TNF-α levels,while Chinchilla rabbits for pyrogen reaction,and NIH mice for tumor-inhibiting effect.Results Compared with those of heat-inactivated CP,the cell walls of ECPP were thin,while the intracellular structure showed no significant change.The protein,polysaccharide and nucleic acid contents of ECPP decreased significantly as compared with those of CPP（P 〈 0.01）.The spleen index in ECPPⅡ group was 2.96,and the inhibitory effect on Ehrlich ascites tumor was observed,with a survival rate of animals of 80%.The rabbits in ECPPⅡ group were qualified in pyrogen test.Abnormal toxicity test showed that the bodyweights of mice in ECPP Ⅱ group increased significantly as compared with those in CPP group.However,the serum IL-2,IFNγ and TNF-α levels of mice in ECPP Ⅱ group increased significantly.Conclusion ECPP decreased the adverse reaction and enhanced the antitumor function,which showed good antitumor effect and might be used as a novel immunomodulator of good prospect of application.