可缓解食物过敏的热灭活乳酸菌的筛选及其全基因组分析
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(东北农业大学食品学院 乳品科学教育部重点实验室 哈尔滨 150030)

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黑龙江省自然科学基金联合引导项目(LH2023C033)


Screening of Heat-killed Lactic Acid Bacteria with Food Allergy Alleviating Effects and Its Whole Genome Sequencing Analysis
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(Key Lab of Dairy Science, Ministry of Education, College of Food Science, Northeast Agricultural University,Harbin 150030)

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    摘要:

    食物过敏(FA)是指人体免疫系统对特定食物产生的异常反应,会导致消化道、呼吸道、皮肤和心血管系统等多个系统的不适。本文通过透明质酸酶抑制试验和RBL-2H3细胞试验,从20株乳酸菌中筛选出抗过敏活性较强的热灭活副干酪乳杆菌JY56。随后,通过全基因组测序和体外安全性试验,评价副干酪乳杆菌Y56菌株的安全性。最后,构建卵清蛋白(OVA)诱导的FA小鼠模型,明确HK-副干酪乳杆菌JY56对FA小鼠临床症状的缓解效果。结果表明,HK-副干酪乳杆菌JY56具有较高的β-氨基己糖苷酶(β-HEX)抑制率,其干预后细胞因子IL-4、IL-5、IL-13、TNF-α的含量显著降低。全基因组学测序分析发现副干酪乳杆菌JY56不具有毒性和传递接收抗生素耐药基因的能力。抗生素敏感性试验、溶血试验、有害代谢产物评价等安全性评价试验表明副干酪乳杆菌JY56对常用抗生素具有敏感性,不产生溶血作用和有害代谢产物。上述说明副干酪乳杆菌JY56具有安全性,其灭活菌体可用于后续研究。OVA刺激后会导致小鼠的体质量和直肠温度降低,过敏评分提高,而HK-副干酪乳杆菌JY56干预后可以明显缓解小鼠过敏症状。本研究结果为HK-副干酪乳杆菌JY56用于缓解食物过敏及相关疾病提供理论依据。

    Abstract:

    Food allergy(FA) is an abnormal response of the human immune system to a specific food that causes discomfort in several systems, including the digestive, respiratory, skin and cardiovascular systems. In this study, heat-killed Lactobacillus paracasei JY56 (HK-L. paracasei JY56) with strong anti-allergic activity was screened from 20 strains of lactic acid bacteria by hyaluronidase inhibition assay and RBL-2H3 cell assay. Subsequently, the safety of L. paracasei JY56 was evaluated by whole genome sequencing and in vitro safety assays. Finally, an ovalbumin (OVA)-induced FA mouse model was constructed to further clarify the effect of HK-L. paracasei JY56 on the alleviation of clinical symptoms in FA mice. The results showed that HK-L. paracasei JY56 had high β-aminohexosidase (β-HEX) inhibition rate, and the levels of cytokines IL-4, IL-5, IL-13 and TNF-α were significantly reduced after its intervention. Whole genome sequencing analysis revealed that L. paracasei JY56 does not possess the ability to virulently and transmitly receive antibiotic resistance genes. The safety evaluation experiments such as antibiotic susceptibility test, hemolysis test and harmful metabolite evaluation revealed that L. paracasei JY56 was susceptible to commonly used antibiotics and did not produce hemolysis or harmful metabolites. OVA stimulation resulted in decreased body weight and rectal temperature and increased allergy score in mice, while HK-L. paracasei JY56 intervention significantly alleviated allergy symptoms in mice. In conclusion, this study provides a theoretical basis for the use of HK-L. paracasei JY56 for the alleviation of food allergy and related diseases.

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郑淼,何梦丽,方如雪,满朝新,张宇,杨鑫焱,赵倩玉,姜毓君.可缓解食物过敏的热灭活乳酸菌的筛选及其全基因组分析[J].中国食品学报,2024,24(9):141-154

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  • 收稿日期:2023-09-02
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  • 在线发布日期: 2024-10-21
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