α-Chaconine is the most abundant glycoalkaloid in potato and toxic to the animal digestive system, but the mechanisms underlying the toxicity are unclear. In this study, mouse small intestinal epithelial cells were incubated with α-chaconine at 0, 0.4, and 0.8 μg/mL for 24, 48, and 72 h to examine apoptosis, mechanical barrier function, and antioxidant ability of the cells using a cell metabolic activity assay, flow cytometry, Western blot, immunofluorescence, and fluorescence quantitative PCR. The results showed that α-chaconine significantly decreased cell proliferation rate, increased apoptosis rate, decreased transepithelial electrical resistance (TEER) value, and increased alkaline phosphatase (AKP) and lactate dehydrogenase (LDH) activities, and there were interactions between α-chaconine concentration and incubation time. α-Chaconine significantly reduced the relative and mRNA expressions of genes coding tight junction proteins zonula occludens-1 (ZO-1) and occludin, increased malondialdehyde (MDA) content, decreased total glutathione (T-GSH) content, reduced the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and γ-glutamylcysteine synthetase (γ-GCS) and the mRNA expressions of SOD, CAT, GSH-Px, and γ-GCS genes. In conclusion, α-chaconine disrupts the cell cycle, destroys the mechanical barrier and permeability of mucosal epithelium, inhibits cell proliferation, and accelerates cell apoptosis.

中文翻译：

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α-查茄碱影响小鼠小肠上皮细胞的凋亡，机械屏障功能和抗氧化能力

α-查茄碱是马铃薯中最丰富的生物碱，对动物的消化系统有毒性，但其毒性机理尚不清楚。在这项研究中，将小鼠小肠上皮细胞与α-查茄碱分别以0、0.4和0.8μg/ mL孵育24、48和72小时，以检查细胞的凋亡，机械屏障功能和抗氧化能力。代谢活性测定，流式细胞仪，蛋白质印迹，免疫荧光和荧光定量PCR。结果表明，α-查茄碱浓度显着降低了细胞增殖速率，增加了细胞凋亡率，降低了上皮电阻（TEER）值，并增加了碱性磷酸酶（AKP）和乳酸脱氢酶（LDH）的活性，并且α-查茄碱浓度之间存在相互作用和潜伏时间。α-Chaconine显着降低紧密连接蛋白zonula occludens-1（ZO-1）和occludin编码基因的相对表达和mRNA表达，丙二醛（MDA）含量增加，总谷胱甘肽（T-GSH）含量降低，过氧化物活性降低歧化酶（SOD），过氧化氢酶（CAT），谷胱甘肽过氧化物酶（GSH-Px）和γ-谷氨酰半胱氨酸合成酶（γ-GCS）以及SOD，CAT，GSH-Px和γ-GCS基因的mRNA表达。总之，α-查茄碱破坏细胞周期，破坏粘膜上皮的机械屏障和通透性，抑制细胞增殖，并加速细胞凋亡。降低了超氧化物歧化酶（SOD），过氧化氢酶（CAT），谷胱甘肽过氧化物酶（GSH-Px）和γ-谷氨酰半胱氨酸合成酶（γ-GCS）的活性以及SOD，CAT，GSH-Px和γ-GCS的mRNA表达基因。总之，α-查茄碱破坏细胞周期，破坏粘膜上皮的机械屏障和通透性，抑制细胞增殖，并加速细胞凋亡。降低了超氧化物歧化酶（SOD），过氧化氢酶（CAT），谷胱甘肽过氧化物酶（GSH-Px）和γ-谷氨酰半胱氨酸合成酶（γ-GCS）的活性以及SOD，CAT，GSH-Px和γ-GCS的mRNA表达基因。总之，α-查茄碱破坏细胞周期，破坏粘膜上皮的机械屏障和通透性，抑制细胞增殖，并加速细胞凋亡。