携带角质细胞生长因子和低氧诱导因子-1α双基因减毒沙门氏菌TPKH的初步安全性评价

Preliminary safety evaluation of attenuated Salmonella TPHK carrying hypoxia inducible factor-1α and keratinocyte growth factor genes

目的:初步评价携带角质细胞生长因子(keratinocyte growth factor,KGF)和低氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)双基因的减毒沙门氏菌(TPKH)的安全性,为其在临床上推广使用提供理论依据。方法:急性毒性试验中,观察不同剂量(1.0×10^8,1.0×10^9,1.0×10^10cfu) TPKH对小鼠精神状态、体重、血液学指标、血清生化指标及脏器指数的影响,同时取高剂量灌胃组的心脏、肝脏、脾脏、肺脏、肾脏及胃肠,制作病理组织切片,观察各脏器病理变化。异常毒性试验中,采用小鼠及豚鼠实验法,小鼠实验中,1.0×10^8cfu TPKH灌胃,观察小鼠的反应及体重变化,共观察7 d;豚鼠实验中,1.0×10^9cfu TPKH灌胃,观察豚鼠的反应及体重变化,共观察7 d。结果:急性毒性试验期间各组小鼠精神状态无异常,各组小鼠体重差异不显著(P> 0.05),各组小鼠的脏器指数及生理生化指标差异不显著(P> 0.05);病理切片结果显示,高剂量组的心脏、肝脏等均无病理性改变。异常毒性试验结果显示,观察期内小鼠及豚鼠均无异常反应,到期后每只小鼠及豚鼠的体重均增加。结论:初步证实了TPKH毒副作用小,安全性好。

Objective: To evaluate the safety of attenuated Salmonella (TPKH) which carries keratinocyte growth factor (KGF) and hypoxia inducible factor-1 α (HIF-1α) genes,so as to provide a theoretical basis for its clinical application.Methods: In the acute toxicological test,the effects of different doses (1.0 × 10^8,1.0 × 10^9,1.0 × 10^10 cfu) of TPKH on the mental state,body weight,hematological indexes,serum biochemical indexes and organ index of mice were observed.Meanwhile,pathological sections were made in the high dose gastric perfusion group to observe the pathological changes of the organs.In the abnormal toxicological test,mice and guinea pig were used.In the mouse test,1.0 × 10^8 cfu TPKH was given by gavage to observe the reaction of mice and the change of body weight for 7 d.In guinea pig test,1.0 × 10^9 cfu TPKH was given by gavage,observing the reaction of mice and the change of body weight for 7 d.Results: During the acute toxicity test period,compared with blank control group,the mental state and body weight of mice in each group had no significant difference (P > 0.05).The organ index,physiological and biochemical indexes showed no significant difference among different groups (P > 0.05).There was no pathological change in the tissue sections.The results of abnormal toxicity test showed that there was no abnormal reaction in mice and guinea pigs during the observation period,and the body weight of each mouse and guinea pig increased after the test.Conclusion: The experiment preliminarily confirmed that TPKH is safe with mild toxicity and side effects.