
基于工程菌EcN表达抗菌免疫肽Scy及其应用
周璐彤, 蒋肖, 靳明亮, 汪以真, 程远之
基于工程菌EcN表达抗菌免疫肽Scy及其应用
Expression of the antimicrobial immunopeptide Scy based on the engineered bacteria EcN and its application
目的 利用大肠杆菌Nissle 1917(EcN)表达抗菌免疫肽Scygonadin(Scy),并评估EcN-Scy缓解结肠炎的作用,为研发新型抗生素替代品提供理论依据。 方法 用PCR扩增载体pnirBMisl(pnir)与抗菌免疫肽Scy目的基因,通过同源重组获得重组载体EcN-Scy。选用20只8周龄雌性BALB/c小鼠,平均分为4组(对照组、DSS组、DSS+EcN组和DSS+EcN-Scy组),每2 d分别灌胃小鼠PBS、EcN菌液或EcN-Scy菌液并记录其体质量,试验期14 d。其中除对照组外,其他3组第0~7天通过在饮水中添加3% DSS诱导结肠炎。第14天麻醉后处死小鼠并采集血清和结肠组织样本进行检测。 结果 与DSS组相比,EcN-Scy处理显著缓解了小鼠体质量下降(P<0.05)和由DSS引起的结肠缩短的症状(P<0.05),明显改善了结肠绒毛结构。与DSS组相比,DSS+EcN-Scy组血清中ALP和Urea水平分别显著升高28.6 U/L、1.35 mol/L(P<0.05);TBil和LDH水平有降低趋势(P>0.05)。与DSS组相比,DSS+EcN-Scy组血清中的促炎因子TNF-α、MCP1和IL-1β水平分别显著降低18.9%、50.9%和8.9%(P<0.05),结肠中MCP1和IL-6分别降低了40.4%和21.9%(P<0.05)。 结论 成功利用工程菌EcN表达抗菌免疫肽Scy,具有显著缓解小鼠结肠炎的效果,能够改善DSS诱导的小鼠体质量下降、结肠缩短及形态破损,并降低血清和结肠炎症因子水平。
Objectives Escherichia coli Nissle 1917 (EcN) was used to express antimicrobial immune peptide Scygonadin (Scy) and the effects of EcN-Scy on alleviating colitis were evaluated to provide theoretical basis for the development of novel substitutes for antibiotic. Methods The vector pnirBMisl (pnir) and the target gene of antimicrobial peptide Scy were amplified with PCR, and the recombinant vector EcN-Scy was obtained by homologous recombination connection. Twenty 8-week-old female BALB/c mice were randomly divided into four groups including the control group, DSS group, DSS+EcN group, and DSS+EcN Scy group. The mice were orally administered with PBS, EcN bacterial solution, or EcN-Scy bacterial solution every 2 days and their body weight was recorded. The experiment lasted for 14 days. Except for the control group, the other three groups were induced colitis by adding 3% DSS to their drinking water from day 0 to day 7. Mice were killed on day 14 after anesthesia, and samples of serum and colon tissue were collected for testing. Results EcN-Scy treatment significantly alleviated the weight loss in mice (P<0.05) and the symptoms of colon shortening caused by DSS (P<0.05), and significantly improved the structure of colonic villi compared with DSS group. The level of ALP and Urea in serum in the DSS+EcN-Scy group was significantly increased by 28.6 U/L and 1.35 mol/L (P<0.05) compared with that in DSS group. The level of TBil and LDH had a decreasing trend (P>0.05). The level of pro-inflammatory cytokines TNF-α, MCP1, and IL-1 β in the serum of the DSS+EcN-Scy group was significantly reduced by 18.9%, 50.9%, and 8.9%, respectively (P<0.05), and the level of MCP1 and IL-6 in the colon were reduced by 40.4% and 21.9%, respectively (P<0.05) compared with that in the DSS group. Conclusions The antimicrobial immune peptide Scy was successfully expressed with the engineered bacterium EcN, which has a significant effect on alleviating colitis in mice. It can improve the weight loss, colon shortening, and morphological damages in mice induced by DSS, and reduce the level of inflammatory factors in serum and colon.
大肠杆菌Nissle 1917 / 抗菌免疫肽Scy / 重组表达 / 结肠炎 / 抗生素替代品研发
Escherichia coli Nissle 1917 (EcN) / antimicrobial immune peptide Scygonadin (Scy) / recombinant expression / colitis / research and development of substitutes for antibiotic
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1 |
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2 |
周雪莼,胡婷婷,王佳慧,等.高效液相色谱-高分辨质谱法快速筛查动物源性药食同源产品中32种抗生素兽药残留[J].吉林中医药,2023,43(12): 1469-1474.
|
3 |
|
4 |
|
5 |
韩菲菲,安沙,谢永刚,等.猪乳铁蛋白肽的分子改良及改良肽的生物学活性研究[J].动物营养学报, 2011, 23(2): 241-249.
|
6 |
|
7 |
|
8 |
|
9 |
|
10 |
|
11 |
|
12 |
苏琰, 李融.抗菌肽的食品保鲜应用及生物合成研究进展[J].食品与机械, 2024,40(7): 208-215.
|
13 |
罗会颖, 姚斌. 理性调控甲醇定向生物合成酵母单细胞蛋白的新策略[J]. 中国科学:生命科学, 2024,54(8): 1488-1490.
|
14 |
李朔, 李潇, 张晓黎, 等. 乳酸菌产γ-氨基丁酸及生物合成技术研究进展[J]. 中国调味品, 2022,47(11): 205-210.
|
15 |
沈晓熳, 杨炽光, 翁晨虹, 等.枯草芽孢杆菌RN8的抗菌活性成分分离及其作用机制[J].中国生物工程杂志, 2024,44(7): 12-28.
|
16 |
|
17 |
郭子好, 方华, 李春源, 等. 锯缘青蟹Scygonadin抗菌免疫肽的毕赤酵母表达及其产物抑菌活性分析[J].饲料研究, 2017(10): 32-36.
|
18 |
KAI L,
|
19 |
田家华, 索小涛, 马晨曦, 等. 溃疡性结肠炎动物模型构建方法的研究进展[J]. 山东医药, 2024,64(13): 111-114.
|
20 |
高梦学, 王丽娜, 黄鹤. 合成生物学在肠道微生态疗法研发中的应用[J]. 合成生物学, 2022, 3(1): 35-52.
|
21 |
卢雯, 刘睿娜, 雷璞, 等.基因工程菌的构建、性能研究及其在疾病诊断和治疗中的应用[J].微生物学报, 2023,63(10): 3758-3772.
|
22 |
|
/
〈 |
|
〉 |