The First Clinical Medical College, Zhejiang Chinese Medical University, Hangzhou, 310053, Zhejiang, China.
The Second Clinical Medical College, Zhejiang Chinese Medical University, Hangzhou, 310053, Zhejiang, China.
Inflammopharmacology. 2024 Aug;32(4):2525-2540. doi: 10.1007/s10787-024-01487-y. Epub 2024 May 17.
Small intestine damage caused by diclofenac is called diclofenac enteropathy. Berberine (BBR), a class of isoquinoline alkaloids derived from Berberis vulgaris and Phellodendron amurense, is widely used in intestinal diseases. The present study evaluated the protective effect of BBR on the intestinal mucosal mechanical barrier in diclofenac enteropathy and its possible action mechanism. The in vitro animal experiment revealed that BBR downregulated the expression of long non-coding RNA H19 (lncRNA H19) in the small intestine and exosomes. In the co-culture experiment involving exosomes and intestinal epithelial cell-6 (IEC-6) cells, the results of qRT-PCR, western blotting, and immunofluorescence assays demonstrated that the elevated expression of lncRNA H19 in the small intestine, conveyed via exosomes derived from the diclofenac group, suppressed the expression levels of autophagy-associated protein 5 (Atg 5) and light chain 3 (LC 3), as well as and the tight junction (TJ) proteins zonula occludens-1 (ZO-1), claudin-1, and occluding, relative to the control group. BBR treatment attenuated exosomal lncRNA H19 levels, upregulated the expression of Atg5 and LC3 expression, enhanced TJ protein expression, and increased the light chain 3 (LC3)-II/LC3-I ratio. These findings significantly elucidated that BBR promoted the restoration of autophagy in IECs by inhibiting exosomal lncRNA H19, thereby mitigating the impairment of the intestinal mucosal mechanical barrier function in diclofenac enteropathy. The process involving exosomal lncRNA H19 regulating autophagy, thereby affecting the intestinal mucosal mechanical barrier, offers a novel perspective for the application of BBR in the treatment of diclofenac enteropathy.
双氯芬酸引起的小肠损伤称为双氯芬酸肠病。小檗碱(BBR)是从小檗属和黄皮树中提取的一类异喹啉生物碱,广泛用于治疗肠道疾病。本研究评估了 BBR 对双氯芬酸肠病肠黏膜机械屏障的保护作用及其可能的作用机制。体外动物实验表明,BBR 下调了小肠和外泌体中长链非编码 RNA H19(lncRNA H19)的表达。在涉及外泌体和肠上皮细胞-6(IEC-6)细胞的共培养实验中,qRT-PCR、western blot 和免疫荧光检测结果表明,双氯芬酸组来源的外泌体中 lncRNA H19 的表达升高,抑制了自噬相关蛋白 5(Atg5)和轻链 3(LC3)的表达水平,以及紧密连接(TJ)蛋白闭锁蛋白-1(ZO-1)、闭合蛋白-1 和封闭蛋白的表达水平,与对照组相比。BBR 处理可降低外泌体 lncRNA H19 水平,上调 Atg5 和 LC3 的表达,增强 TJ 蛋白表达,增加 LC3-II/LC3-I 比值。这些发现表明,BBR 通过抑制外泌体 lncRNA H19 促进 IEC 中的自噬恢复,从而减轻双氯芬酸肠病中肠黏膜机械屏障功能的损害。外泌体 lncRNA H19 调节自噬从而影响肠黏膜机械屏障的过程为 BBR 在治疗双氯芬酸肠病中的应用提供了新的视角。