葡萄果皮提取物对砷致小鼠小肠毒性的缓解作用

赵丹瑜; 仪慧兰

doi:10.13386/j.issn1002-0306.2023040011

葡萄果皮提取物对砷致小鼠小肠毒性的缓解作用

Mitigative Effect of Grape Skin Extract on Arsenic-induced Small Intestinal Toxicity in a Mouse Model

摘要

摘要: 目的：探究砷摄入对小肠的毒性及葡萄果皮提取物（grape skin extract，GSE）的干预效应。方法：模拟人类饮水砷暴露，小鼠饮用浓度10 mg/L的砷溶液56 d，建立小鼠砷中毒模型，并用不同浓度GSE隔天灌胃干预（150 mg/kg bw和300 mg/kg bw）；取小鼠小肠组织，显微镜观察组织形态结构；试剂盒法检测还原型谷胱甘肽（glutathione，GSH）、丙二醛（malondialdehyde，MDA）和H₂O₂含量，及总超氧化物歧化酶（total superoxide dismutase，T-SOD）活性；qRT-PCR检测紧密连接基因和炎症通路IL-6/JAK2/STAT-3基因表达水平。结果：砷染毒后，小鼠小肠绒毛变短、排列紊乱，黏膜固有层及黏膜下层大量炎细胞浸润；小肠组织的GSH含量和T-SOD活性分别降低17.1%和25.2%，MDA和H₂O₂含量分别增加68.8%和54.3%（P<0.05）；细胞紧密连接基因ZO-1、ZO-2、occludin、claudin-1和claudin-7显著下调表达（P<0.05）；炎症通路IL-6/JAK2/STAT-3基因转录水平显著增高（P<0.05）。GSE高浓度干预组（300 mg/kg bw），小肠黏膜损伤减轻，肠绒毛趋于正常，炎症浸润减轻；与砷染毒组比较，GSH含量增加17.9%、T-SOD活性升高14.3%、MDA和H₂O₂含量分别减少33.8%和25.4%（P<0.05）；紧密连接基因显著上调表达（P<0.05）；炎症通路IL-6/JAK2/STAT-3基因转录显著降低（P<0.05）。GSE低浓度干预（150 mg/kg bw）对砷毒性有一定的缓解作用，但差异不显著，无统计学意义（P>0.05）。结论：GSE通过抑制砷暴露诱导的小肠组织氧化应激、炎症反应及功能基因转录下调，缓解砷的小肠毒性，对机体具有保护作用。

Abstract: Objective: To investigate the arsenic-induced small intestinal toxicity and the protective effect of grape skin extract (GSE) against arsenic toxicity. Methods: The small intestinal toxicity was induced by 10 mg/L arsenic via drinking water for 56 days, and was intervened with GSE (150 mg/kg bw and 300 mg/kg bw) by gavage every other day in mice. Small intestine tissue samples of mice were collected and observed by microscope. Glutathione (GSH), malondialdehyde (MDA) and H₂O₂ contents, as well as total superoxide dismutase (T-SOD) were determined by using commercial kits. qRT-PCR was used to detect expression levels of the tight junction genes and the inflammatory pathway IL-6/JAK2/STAT-3 genes. Results: The results showed that 56 days exposure to 10 mg/L arsenic via drinking water resulted in shortened and disordered intestinal villi, with large numbers of inflammatory cells infiltrating the mucosa propria and submucosa. GSH content and T-SOD activity decreased by 17.1% and 25.2%, while MDA and H₂O₂ contents increased by 68.8% and 54.3%, respectively (P<0.05) in the small intestinal tissue of arsenic-treated mice. The mRNA levels of IL-6, JAK2 and STAT-3 were upregulated in the small intestinal tissue of mice exposed to arsenic (P<0.05). Meanwhilethe mRNA levels of the ZO-1, ZO-2, occludin, claudin1 and claudin7 genes, which encode the key components of tight junction (TJ) complexes, were downregulated (P<0.05). However, the application of GSE (300 mg/kg bw) significantly alleviated the damage and inflammatory infiltration in small intestine. Compare to the As group, GSH content and T-SOD activity increased by 17.9% and 14.3%. MDA and H₂O₂ contents decreased by 33.8% and 25.4% (P<0.05). Arsenic-mediated gene expression in the IL-6/JAK2/STAT3 pathway was down-regulated (P<0.05). Moreover, the arsenic-induced down-regulation of TJ genes were markedly relieved in the As+GSE (300 mg/kg bw) group (P<0.05). The As+GSE (150 mg/kg bw) group had a certain alleviating effect on arsenic toxicity, but the difference had no statistical significance (P>0.05). Conclusion: The application of GSE provides significant protection against arsenic-induced small intestinal toxicity by attenuating the oxidative stress and inflammatory responses, and inhibiting the down-regulation of some functional genes.