Hung Ying-Kai, Ho Shang-Tse, Kuo Ching-Yun, Chen Ming-Ju
Department of Animal Science and Technology, National Taiwan University, Taipei 106, Taiwan.
Department of Wood Based Materials and Design, National Chiayi University, Chiayi 600, Taiwan.
Int J Med Sci. 2021 Feb 18;18(8):1778-1785. doi: 10.7150/ijms.53599. eCollection 2021.
The mucus integrity and abnormal inflammatory response are the crucial biomarker of inflammatory bowel disease (IBD). Velvet antler (VA) has been used as traditional Chinese medicines for many years. Anti-inflammatory property was demonstrated suppression of cyclooxygenase-2 and cytokines protein expression. And it has further proved to promote wound healing in streptozotocin-induced diabetic rats model. The aforementioned functionalities of VA extracts may be associated with the treatment of IBD. Thus, the aim of present study was to evaluate the effect of velvet antler water extracts form Formosan Sambar deer (, SVAE) and red deer (, RVAE) on the barrier function and to investigate the possible mechanism using model. Human colonic epithelial cell models (Caco-2) were co-cultured with various concentrations of both SVAE and RVAE (250-500 µg mL) in dextran sulfate sodium (DSS)-induced colitis model. Trans-epithelial electrical resistance (TEER) value and the macromolecule permeability of Fluorescein isothiocyanate (FITC)-labeled dextran were measured to evaluate the integrity of monolayer of Caco-2. Western blotting was performed for analysis of protein expressions of occludin, Zonula occludens-1 (ZO-1), claudin-1, claudin-2 and myosin light chain kinase (MLCK). The cytotoxicity was conducted by MTT assay. Results indicated that both SVAE and RVAE could enhance integrity of monolayer in dextran sulfate sodium (DSS)-induced colonic epithelial cell model (Caco-2) through reducing the decline of trans-epithelial electrical resistance (TEER) and macromolecule permeability at the concentration of 250 μg mL. RVAE significantly increased the expression of tight junction proteins (occludin and ZO-1) while SVAE significantly reduced the activity of MLCK ( < 0.05.). Elevated C-C chemokine ligand 20 (CCL20) production suggested that both SVAE and RVAE could enhance the repair of epithelial cell. Besides, MTT assay revealed that both extracts showed no cytotoxicity. Thus, SVAE and RVAE supplementation may attenuate barrier damage by enhancing the occludin and ZO-1 protein expression, decreasing MLCK expression, promoting the CCL20 production. In the future, animal study is needed for further confirmation.
黏液完整性和异常炎症反应是炎症性肠病(IBD)的关键生物标志物。鹿茸(VA)作为传统中药已使用多年。其抗炎特性表现为对环氧合酶 - 2和细胞因子蛋白表达的抑制。并且在链脲佐菌素诱导的糖尿病大鼠模型中进一步证明它能促进伤口愈合。VA提取物的上述功能可能与IBD的治疗有关。因此,本研究的目的是评估台湾水鹿鹿茸水提取物(SVAE)和马鹿鹿茸水提取物(RVAE)对屏障功能的影响,并使用模型研究其可能的机制。在葡聚糖硫酸钠(DSS)诱导的结肠炎模型中,将人结肠上皮细胞模型(Caco - 2)与不同浓度(250 - 500μg/mL)的SVAE和RVAE共同培养。测量跨上皮电阻(TEER)值和异硫氰酸荧光素(FITC)标记的葡聚糖的大分子通透性,以评估Caco - 2单层的完整性。进行蛋白质印迹分析紧密连接蛋白occludin、紧密连接蛋白 - 1(ZO - 1)、闭合蛋白 - 1、闭合蛋白 - 2和肌球蛋白轻链激酶(MLCK)的表达。通过MTT法进行细胞毒性检测。结果表明,在250μg/mL浓度下,SVAE和RVAE均可通过减少跨上皮电阻(TEER)的下降和大分子通透性来增强葡聚糖硫酸钠(DSS)诱导的结肠上皮细胞模型(Caco - 2)中单层的完整性。RVAE显著增加紧密连接蛋白(occludin和ZO - 1)的表达,而SVAE显著降低MLCK的活性(P < 0.05)。C - C趋化因子配体20(CCL20)产量的升高表明SVAE和RVAE均可增强上皮细胞的修复。此外,MTT法显示两种提取物均无细胞毒性。因此,补充SVAE和RVAE可能通过增强occludin和ZO - 1蛋白表达、降低MLCK表达、促进CCL20产生来减轻屏障损伤。未来,需要进行动物研究以进一步证实。