吴茱萸碱维持结肠癌上皮细胞的屏障完整性的机制探讨
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摘要
目的观察吴茱萸碱对LPS诱导结肠癌上皮细胞(Caco-2)炎症反应的保护作用及机制。方法采用LPS处理人结肠癌上皮细胞(Caco-2)制备肠上皮细胞炎症损伤模型,用适宜浓度的吴茱萸碱共培养细胞,通过短路电流检测跨上皮电阻、小室共培养检测FD4透过率测定细胞旁通透性,采用ELISA检测上清液中IL-1β和TNF-α含量,通过蛋白印记检测occludin、accludin-1和ZO-1的表达。结果与LPS处理组相比,吴茱萸碱刺激后显著增加跨上皮电阻,降低细胞旁通透性。与对照组相比,LPS处理显著增加IL-1β和TNF-α含量,而吴茱萸碱能逆转促炎症因子的表达。紧密连接相关蛋白occludin、accludin-1和ZO-1在LPS处理后显著降低,而吴茱萸碱处理后,紧密连接蛋白表达量显著增加。结论吴茱萸碱通过降低炎症反应,增加紧密连接蛋白表达,维持肠上皮细胞的屏障完整性。
Objective To observe the protective effect and mechanism of evodiamine on LPS-induced inflammatory response of colon cancer epithelial cells(Caco-2). Methods Human colon cancer epithelial cells(Caco-2) were treated with LPS to prepare intestinal epithelial inflammatory injury model. Cells were co-cultured with appropriate concentration of evodiamine. The short-circuit current detection was used to detect FD4 transmittance by transepithelial electrical resistance and co-culture. Peripheral permeability,tdheet elcetveedl s boyf lWLe-s1 tβe rann db lTotNtiFn-ga. iRn etsheu lstusp e rCnoatmapnta rwede r ew idteht etchtee dL bPyS-EtrLeIaSteAd, agrnod utph,e eevxopdrieassmiionnes soifg noicfcilcuadnitnl,y a icncclruedaisne-d1 tarnadn sZe0 p-i1 t hewleirael electrical resistance and decreased paracellular permeability. Compared with the control group, LPS treatment significantly increased the levels of lL-1β and TNF-a, while evodiamine reversed the expression of pro-inflammatory factors. The tight junction-associated proteins occludin, accludin-1, and Z0-1 were significantly reduced after LPS treatment, whereas the expression of tight junction protein was significantly increased after evodiamine treatment. Conclusion Evodiamine can maintain the barrier integrity of intestinal epithelial cells by reducing the inflammatory response and increasing the expression of tight junction proteins.
Objective To observe the protective effect and mechanism of evodiamine on LPS-induced inflammatory response of colon cancer epithelial cells(Caco-2). Methods Human colon cancer epithelial cells(Caco-2) were treated with LPS to prepare intestinal epithelial inflammatory injury model. Cells were co-cultured with appropriate concentration of evodiamine. The short-circuit current detection was used to detect FD4 transmittance by transepithelial electrical resistance and co-culture. Peripheral permeability,tdheet elcetveedl s boyf lWLe-s1 tβe rann db lTotNtiFn-ga. iRn etsheu lstusp e rCnoatmapnta rwede r ew idteht etchtee dL bPyS-EtrLeIaSteAd, agrnod utph,e eevxopdrieassmiionnes soifg noicfcilcuadnitnl,y a icncclruedaisne-d1 tarnadn sZe0 p-i1 t hewleirael electrical resistance and decreased paracellular permeability. Compared with the control group, LPS treatment significantly increased the levels of lL-1β and TNF-a, while evodiamine reversed the expression of pro-inflammatory factors. The tight junction-associated proteins occludin, accludin-1, and Z0-1 were significantly reduced after LPS treatment, whereas the expression of tight junction protein was significantly increased after evodiamine treatment. Conclusion Evodiamine can maintain the barrier integrity of intestinal epithelial cells by reducing the inflammatory response and increasing the expression of tight junction proteins.
引文
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[2]Ye D,Lei W,Al-Sadi R,et al.990 IL-6 Induced Increase in Intestinal Epithelial Tight Junction Permeability is Mediated by AP-1 Induced UpRegulation of Claudin-2 Gene Activity[J].Gastroenterology,2012,142(5):S-175-S-75.
[3]Ohland CL,Macnaughton WK.Probiotic bacteria and intestinal epithelial barrier function[J].Am J Physiol Gastrointest Liver Physiol,2010,298(6):807-19.
[4]Xavier RJ,Podolsky DK.Unravelling the pathogenesis of inflammatory bowel disease[J].Nature,2007,448(7152):427-34.
[5]曹阳,高玉华,于明新.吴茱萸碱对急性放射性肠炎疗效及相关机制实验研究[J].辽宁中医药大学学报,2010,(02):31-33.
[6]裘杨溢,徐盛涛,徐进宜.吴茱萸碱类衍生物的生物活性研究进展[J].药学与临床研究,2017,25(3):221-26.
[7]Hong-Wei,Zhao,Yue-Hong,et al.Effect of toll-like receptor 3 agonist poly I:C on intestinal mucosa and epithelial barrier function in mouse models of acute colitis[J].World Journal of Gastroenterology,2017,23(6):999-1009.
[8]Oshitani N,Watanabe K,Nakamura S,et al.Dislocation of tight junction proteins without F-actin disruption in inactive Crohn’s disease[J].International Journal of Molecular Medicine,2005,15(3):407-10.
[9]Yamamoto-Furusho JK,Santiago-Hernández JJ,Pérez-Hernández N,et al.Interleukin 1β(IL-1B)and IL-1 antagonist receptor(IL-1RN)gene polymorphisms are associated with the genetic susceptibility and steroid dependence in patients with ulcerative colitis[J].Journal of Clinical Gastroente rology,2011,45(6):531.
[10]Jiao YF,Lu M,Zhao YP,et al.N-Methylcytisine Ameliorates DextranSulfate-Sodium-Induced Colitis in Mice by Inhibiting the Inflammatory Response[J].Molecules,2018,23(3):510.
[11]Shen P,Zhang Z,He Y,et al.Magnolol treatment attenuates dextran sulphate sodium-induced murine experimental colitis by regulating inflammation and mucosal damage[J].Life Sciences,2018,196:69-76.