Department of General Surgery, Jinling Hospital, Second Military Medical University, Nanjing, China.
Inflammation. 2012 Jun;35(3):841-9. doi: 10.1007/s10753-011-9385-6.
Endoplasmic reticulum stress (ER stress) in intestinal epithelial cells (IECs) plays an important role in the pathogenesis and perpetuation of inflammatory bowel disease (IBD). The aim of this study is to explore the potential of berberine (BBR) in regulating pro-inflammatory cytokine-induced ER stress and apoptosis in IECs. ER stress in cultured Caco-2 cells was induced by IFN-γ/TNF-α and tunicamycin, respectively. The experimental groups were pretreated with BBR. Cell viability was determined by MTT assay. In vitro apoptosis was examined by flow cytometry using annexin V-FITC labeling. The molecular markers of ER stress, including GRP-78, p-JNK, caspase-12, and cleaved caspase-3 were analyzed by Western blot. Xbp-1 mRNA splicing was detected by RT-PCR. Pretreatment of BBR helped to survive in cytokine-induced Caco-2 cells. BBR significantly attenuated cytokine-induced Caco-2 apoptosis. GRP78 expression and xbp-1 mRNA splicing were enhanced significantly in the presence of IFN-γ/TNF-α and tunicamycin, and they could be dampened by BBR. Further study revealed BBR could downregulate JNK phosphorylation, and the level of caspase-12 and cleaved caspase-3. Berberine can ameliorate pro-inflammatory cytokines induced ER stress in vitro, and it might be one of the targeted therapeutic agents for IBD.
内质网应激（ER 应激）在肠上皮细胞（IECs）中在炎症性肠病（IBD）的发病机制和持续存在中起着重要作用。本研究旨在探讨小檗碱（BBR）在调节 IECs 中促炎细胞因子诱导的 ER 应激和细胞凋亡中的潜力。用 IFN-γ/TNF-α和衣霉素分别诱导培养的 Caco-2 细胞中的 ER 应激。实验组用 BBR 预处理。通过 MTT 测定法测定细胞活力。通过使用 Annexin V-FITC 标记通过流式细胞术检查体外细胞凋亡。通过 Western blot 分析 ER 应激的分子标志物，包括 GRP-78、p-JNK、caspase-12 和 cleaved caspase-3。通过 RT-PCR 检测 Xbp-1 mRNA 剪接。BBR 的预处理有助于在细胞因子诱导的 Caco-2 细胞中存活。BBR 显著减轻了细胞因子诱导的 Caco-2 细胞凋亡。IFN-γ/TNF-α和衣霉素存在时，GRP78 表达和 xbp-1 mRNA 剪接显著增强，BBR 可减弱其表达。进一步的研究表明，BBR 可以下调 JNK 磷酸化，以及 caspase-12 和 cleaved caspase-3 的水平。小檗碱可以改善体外促炎细胞因子诱导的 ER 应激，它可能是 IBD 的靶向治疗药物之一。
