Department of Oncological Radiotherapy, Affiliated Huai'an First Hospital, Nanjing Medical University, Huaian City, Jiangsu Province, PR China.
Department of Gastrointestinal Surgery, Affiliated Huai'an First Hospital, Nanjing Medical University, Huaian City, Jiangsu Province, PR China.
Life Sci. 2019 Sep 15;233:116697. doi: 10.1016/j.lfs.2019.116697. Epub 2019 Jul 25.
The present study investigated if berberine might induce Zrt-Irt-like protein 14 (ZIP14) and affect zinc redistribution to protect intestinal barrier in sepsis.
Rodent model of sepsis was induced by cecal ligation and puncture (CLP). Plasma endotoxin was assayed by LAL test and plasma zinc was measured by flame atomic spectrophotometer. Gut mucosal permeability was determined by plasma FITC-dextran. Zinc content and ZIP14 mRNA in gut mucosa were assayed by spectrophotometer and qRT-PCR, respectively. Tight junction integrity of Caco-2 was evaluated by transepithelial electrical resistance (TEER). Tight junction (TJ) protein expression was detected by Western blotting.
Berberine and zinc gluconate pretreatment to CLP rats improved survival rate, reduced plasma endotoxin level, alleviated hypozincemia, increased zinc accumulation and ZIP14 mRNA expression in the intestinal mucosa. Berberine and zinc gluconate pretreatment decreased CLP-elicited intestinal hyperpermeability to FITC-dextran. These effects of berberine in vivo were abolished by AG1024. In vitro, lipopolysaccharide (LPS) repressed zinc transfer into Caco-2 cells exposed to zinc gluconate. Berberine and IGF-I treatment increased ZIP14 protein expression and promoted zinc transfer into Caco-2 cells exposed to zinc gluconate plus LPS. Berberine treatment induced TJ protein (claudin-1 and occludin) and raised TEER in LPS-treated Caco-2 cells. These effects of berberine in vitro were partially inhibited by ZIP14 siRNA.
The present study reveals that berberine induces ZIP14 expression and affects zinc re- distribution to protect intestinal barrier in sepsis, which is partially linked with the activation of IGF-I signaling.
本研究旨在探讨小檗碱是否可以通过诱导 Zrt-Irt 样蛋白 14（ZIP14）来影响锌的重新分布，从而在脓毒症中保护肠道屏障。
采用盲肠结扎穿孔（CLP）法建立脓毒症大鼠模型。采用 LAL 试验检测血浆内毒素，火焰原子分光光度法检测血浆锌含量，FITC-右旋糖酐法检测肠道黏膜通透性。采用分光光度计和 qRT-PCR 分别检测肠道黏膜锌含量和 ZIP14mRNA 表达。通过跨上皮电阻（TEER）评估 Caco-2 紧密连接的完整性，通过 Western blot 检测紧密连接（TJ）蛋白的表达。
小檗碱和葡萄糖酸锌预处理 CLP 大鼠可提高存活率，降低血浆内毒素水平，缓解低锌血症，增加肠道黏膜锌蓄积和 ZIP14mRNA 表达。小檗碱和葡萄糖酸锌预处理可降低 CLP 诱导的肠道对 FITC-右旋糖酐的高通透性。AG1024 可消除小檗碱的这些体内作用。在体外，脂多糖（LPS）抑制锌向暴露于葡萄糖酸锌的 Caco-2 细胞的转移。小檗碱和 IGF-I 处理可增加 ZIP14 蛋白表达，并促进锌向暴露于 LPS 加葡萄糖酸锌的 Caco-2 细胞的转移。小檗碱处理可诱导 LPS 处理的 Caco-2 细胞中 TJ 蛋白（claudin-1 和 occludin）和升高 TEER。小檗碱在体外的这些作用部分被 ZIP14 siRNA 抑制。
本研究表明，小檗碱通过诱导 ZIP14 表达和影响锌的重新分布来保护脓毒症中的肠道屏障，这与 IGF-I 信号的激活部分相关。
