摘要
氧化应激是造成溃疡性结肠炎(UC)的重要原因之一,而超氧化物歧化酶(SOD)是调控氧化应激的基本分子。为了探究TAT-SOD脂质体对UC模型大鼠的口服治疗作用,作者利用乙醇沉淀和大孔树脂分离的方法,从基因重组毕赤酵母菌发酵液中分离纯化TAT-SOD;利用TritonX-114相分离法,去除其细菌内毒素;利用逆向蒸发法,制备TAT-SOD脂质体;利用DSS诱导构建UC模型大鼠,经灌胃给药,通过DAI评分、结肠密度、组织切片、血清SOD酶活、血清脂多糖含量,评价其治疗效果。结果表明:初步分离获得TAT-SOD,比活为11 003 U/mg;TritonX-114对内毒素的去除率为99.30%;成功制备TAT-SOD脂质体,经体外模拟消化液处理,其酶活回收率为81.80%;动物实验中,SOD给药组与模型组相比,DAI评分均显著降低(P<0.05),且TAT-SOD脂质体组的评分最好;结肠密度和组织切片观察发现,结肠组织损伤情况均明显改善,且TAT-SOD脂质体组的改善最好;血清中SOD酶活均显著提高(P<0.05),且TATSOD脂质体组的酶活最高;血清中LPS含量无显著差异。这些表明,TAT-SOD脂质体对DSS诱导UC模型大鼠有良好的口服治疗效果,有望为UC的辅助治疗提供新的选择。
Oxidative stress is one of the major causes of ulcerative colitis(UC),and superoxide dismutase(SOD)is the basic molecule to regulate oxidative stress.This study investigated the therapeutic effect of oral administered TAT-SOD liposome(L-TAT-SOD)on UC rat models.The isolated and purified TAT-SOD from the fermentation broth of recombinant Pichia pastoris was achieved by ethanol precipitation,macroporous resin column chromatography and TritonX-114 phase separation.Subsequently,the L-TAT-SOD was prepared by reverse evaporation.Finally,the UC rat models were established by dextran sodium sulfate(DSS)induction and administered by intragastric administration.The anti-UC effect of L-TAT-SOD was evaluated by DAI score,colon density, damage of colon tissue, serum SOD enzyme activity and serum lipopoly saccharide content.Results showed that: The specific activity of isolated TAT-SOD was 11 003 U/mg. The recovery rates of enzyme activity of successfully prepared L-TAT-SOD were about 80% and 90% in simulated gastric and intestinal juice, respectively. In anti-UC experiments, all administered groups had lower DAI scores(P<0.05) than that of the model group. In contrast, the L-TAT-SOD group had the lowest DAI score.The results of colon density and tissue section showed that the damaged colon tissue was significantly repaired in all administered groups, and the L-TAT-SOD group performed best.Meanwhile, the serum SOD enzyme activity was significantly increased in all groups(P<0.05), and the activity in L-TAT-SOD group was highest. However, there was no significant difference in serum lipopolysaccharide content in each group. The oral administered L-TAT-SOD is confirmed with a beneficial therapeutic effect on DSS-induced UC rat models, expected to be a new option for the adjuvant therapy of UC.
作者
周建森
周秋婷
叶芬
林春通
刘航琪
刘树滔
ZHOU Jiansen;ZHOU Qiuting;YE Fen;LIN Chuntong;LIU Hangqi;LIU Shutao(Institute of Biotechnology,Fuzhou University,Fuzhou 350002,China)
出处
《食品与生物技术学报》
CAS
CSCD
北大核心
2020年第10期26-33,共8页
Journal of Food Science and Biotechnology
基金
国家重点研发计划专项项目(2016YFD0400202)
福建省科技厅产业技术开发与应用计划:区域发展项目(2018N3009)
福州市高校产学研合作项目(2017-G-104)。
关键词
超氧化物歧化酶
脂质体
结肠炎
抗氧化
superoxide dismutase
liposomes
ulcerative colitis
antioxidant
