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转录组学、16S 核糖体核糖核酸和网络药理学分析揭示了绿茶多酚对乌蒙山黑骨鸡感染柔嫩艾美耳球虫的抗球虫作用机制。

Transcriptomic, 16S ribosomal ribonucleic acid and network pharmacology analyses shed light on the anticoccidial mechanism of green tea polyphenols against Eimeria tenella infection in Wuliangshan black-boned chickens.

机构信息

Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, 650201, Yunnan, People's Republic of China.

Key Laboratory of Veterinary Public Health of Yunnan Province, College of Veterinary Medicine, Yunnan Agricultural University, Kunming, 650201, Yunnan, People's Republic of China.

出版信息

Parasit Vectors. 2023 Sep 19;16(1):330. doi: 10.1186/s13071-023-05922-x.

Abstract

BACKGROUND

Eimeria tenella is an obligate intracellular parasitic protozoan that invades the chicken cecum and causes coccidiosis, which induces acute lesions and weight loss. Elucidating the anticoccidial mechanism of action of green tea polyphenols could aid the development of anticoccidial drugs and resolve the problem of drug resistance in E. tenella.

METHODS

We constructed a model of E. tenella infection in Wuliangshan black-boned chickens, an indigenous breed of Yunnan Province, China, to study the efficacy of green tea polyphenols against the infection. Alterations in gene expression and in the microbial flora in the cecum were analyzed by ribonucleic acid (RNA) sequencing and 16S ribosomal RNA (rRNA) sequencing. Quantitative real-time polymerase chain reaction was used to verify the host gene expression data obtained by RNA sequencing. Network pharmacology and molecular docking were used to clarify the interactions between the component green tea polyphenols and the targeted proteins; potential anticoccidial herbs were also analyzed.

RESULTS

Treatment with the green tea polyphenols led to a reduction in the lesion score and weight loss of the chickens induced by E. tenella infection. The expression of matrix metalloproteinase 7 (MMP7), MMP1, nitric oxide synthase 2 and ephrin type-A receptor 2 was significantly altered in the E. tenella infection plus green tea polyphenol-treated group and in the E. tenella infection group compared with the control group; these genes were also predicted targets of tea polyphenols. Furthermore, the tea polyphenol (-)-epigallocatechin gallate acted on most of the targets, and the molecular docking analysis showed that it has good affinity with interferon induced with helicase C domain 1 protein. 16S ribosomal RNA sequencing showed that the green tea polyphenols had a regulatory effect on changes in the fecal microbiota induced by E. tenella infection. In total, 171 herbs were predicted to act on two or three targets in MMP7, MMP1, nitric oxide synthase 2 and ephrin type-A receptor 2.

CONCLUSIONS

Green tea polyphenols can directly or indirectly regulate host gene expression and alter the growth of microbiota. The results presented here shed light on the mechanism of action of green tea polyphenols against E. tenella infection in chickens, and have implications for the development of novel anticoccidial products.

摘要

背景

柔嫩艾美耳球虫是一种专性细胞内寄生的原生动物,侵袭鸡的盲肠并引起球虫病,导致急性病变和体重减轻。阐明绿茶多酚的抗球虫作用机制有助于开发抗球虫药物并解决柔嫩艾美耳球虫的耐药性问题。

方法

我们构建了中国云南省乌蒙山黑骨鸡柔嫩艾美耳球虫感染模型,研究绿茶多酚对感染的疗效。通过核糖核酸(RNA)测序和 16S 核糖体 RNA(rRNA)测序分析了盲肠中基因表达和微生物菌群的变化。采用实时定量聚合酶链反应验证 RNA 测序获得的宿主基因表达数据。网络药理学和分子对接用于阐明成分绿茶多酚与靶向蛋白之间的相互作用;还分析了潜在的抗球虫草药。

结果

绿茶多酚治疗可降低柔嫩艾美耳球虫感染诱导的鸡病变评分和体重减轻。与对照组相比,感染后加绿茶多酚组和感染组鸡的基质金属蛋白酶 7(MMP7)、MMP1、一氧化氮合酶 2 和 Ephrin 型-A 受体 2 的表达明显改变;这些基因也是茶多酚的预测靶点。此外,茶多酚(-)-表没食子儿茶素没食子酸酯作用于大多数靶点,分子对接分析表明它与具有螺旋酶 C 结构域 1 蛋白的干扰素具有良好的亲和力。16S rRNA 测序显示,绿茶多酚对柔嫩艾美耳球虫感染引起的粪便微生物群变化具有调节作用。共有 171 种草药被预测作用于 MMP7、MMP1、一氧化氮合酶 2 和 Ephrin 型-A 受体 2 中的两个或三个靶点。

结论

绿茶多酚可以直接或间接调节宿主基因表达并改变微生物群的生长。本研究结果揭示了绿茶多酚对鸡柔嫩艾美耳球虫感染的作用机制,为开发新型抗球虫产品提供了依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2555/10510215/fab1831e4043/13071_2023_5922_Fig1_HTML.jpg

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