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Matrine reduced intestinal stem cell damage in eimeria necatrix-infected chicks via blocking hyperactivation of Wnt signaling
Phytomedicine ( IF 6.7 ) Pub Date : 2024-01-15 , DOI: 10.1016/j.phymed.2024.155363
Geng-Xiu Zan 1 , Xiao-Fan Wang 1 , Shao-Kang Yan 1 , Ying-Chao Qin 1 , Lang-Qun Yao 2 , Chun-Qi Gao 1 , Hui-Chao Yan 1 , Jia-Yi Zhou 1 , Xiu-Qi Wang 1
Affiliation  

Coccidiosis is a rapidly spreading and acute parasitic disease that seriously threatening the intestinal health of poultry. Matrine from leguminous plants has anthelmintic and anti-inflammatory properties. This assay was conducted to explore the protective effects of Matrine and the AntiC (a Matrine compound) on Eimeria necatrix (EN)-infected chick small intestines and to provide a nutritional intervention strategy for EN injury. The (chick) experiment: A total of 392 one-day-old yellow-feathered broilers were randomly assigned to six groups in a 21-day study: control group, 350 mg/kg Matrine group, 500 mg/kg AntiC group, EN group, and EN + 350 mg/kg Matrine group, EN + 500 mg/kg AntiC group. The (chick intestinal organoids, IOs): The IOs were treated with PBS, Matrine, AntiC, 3 μM CHIR99021, EN (15,000 EN sporozoites), EN + Matrine, EN + AntiC, EN + Matrine + CHIR99021, EN + AntiC + CHIR99021. The structural integrity of chicks jejunal crypt-villus axis was evaluated by hematoxylin and eosin (H&E) staining and transmission electron microscopy (TEM). And the activity of intestinal stem cells (ISCs) located in crypts was assessed by expansion advantages of a primary in IOs model. Then, the changes of Wnt/β-catenin signaling in jejunal tissues and IOs were detected by Real-Time qPCR,Western blotting and immunohistochemistry. The results showed that dietary supplementation with Matrine or AntiC rescued the jejunal injury caused by EN, as indicated by increased villus height, reduced crypt hyperplasia, and enhanced expression of tight junction proteins. Moreover, there was less budding efficiency of the IOs expanded from jejunal crypts of chicks in the EN group than that in the Matrine and AntiC group, respectively. Further investigation showed that AntiC and Matrine inhibited EN-stimulated Wnt/β-catenin signaling. The fact that Wnt/β-catenin activation via CHIR99021 led to the failure of Matrine and AntiC to rescue damaged ISCs confirmed the dominance of this signaling. Our results suggest that Matrine and AntiC inhibit ISC proliferation and promote ISC differentiation into absorptive cells by preventing the hyperactivation of Wnt/β-catenin signaling, thereby standardizing the function of ISC proliferation and differentiation, which provides new insights into mitigating EN injury by Matrine and AntiC.

中文翻译:


苦参碱通过阻断 Wnt 信号过度激活,减少艾美耳球虫感染雏鸡的肠道干细胞损伤



球虫病是一种传播迅速的急性寄生虫病,严重威胁家禽肠道健康。来自豆科植物的苦参碱具有驱虫和抗炎特性。本试验旨在探讨苦参碱和AntiC(一种苦参碱化合物)对感染艾美耳球虫(EN)的雏鸡小肠的保护作用,并为EN损伤提供营养干预策略。 (小鸡)实验:在为期 21 天的研究中,总共 392 只一日龄黄羽肉鸡被随机分为 6 组:对照组、350 mg/kg 苦参碱组、500 mg/kg AntiC 组、EN组、EN+350mg/kg苦参碱组、EN+500mg/kgAntiC组。 (鸡肠道类器官,IO):用 PBS、苦参碱、AntiC、3 μM C​​HIR99021、EN(15,000 EN 子孢子)、EN + 苦参碱、EN + AntiC、EN + 苦参碱 + CHIR99021、EN + AntiC + CHIR99021 处理 IO 。通过苏木精和伊红 (H&E) 染色和透射电子显微镜 (TEM) 评估雏鸡空肠隐窝绒毛轴的结构完整性。通过 IOs 模型中原代细胞的扩增优势来评估位于隐窝中的肠干细胞 (ISC) 的活性。然后通过Real-Time qPCR、Western blotting和免疫组化检测空肠组织和IOs中Wnt/β-catenin信号的变化。结果表明,饮食中补充苦参碱或 AntiC 可挽救 EN 引起的空肠损伤,表现为绒毛高度增加、隐窝增生减少和紧密连接蛋白表达增强。此外,EN组中从雏鸡空肠隐窝扩增出的IOs的出芽效率分别低于苦参碱组和AntiC组。 进一步研究表明 AntiC 和苦参碱抑制 EN 刺激的 Wnt/β-catenin 信号传导。通过 CHIR99021 激活 Wnt/β-连环蛋白导致苦参碱和 AntiC 无法拯救受损的 ISC,这一事实证实了该信号传导的主导地位。我们的研究结果表明,苦参碱和AntiC通过阻止Wnt/β-catenin信号的过度激活来抑制ISC增殖并促进ISC分化为吸收细胞,从而标准化ISC增殖和分化的功能,这为苦参碱和AntiC减轻EN损伤提供了新的见解。滑稽。
更新日期:2024-01-15
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