Li Shujie, Xie Ruijin, Jiang Chengrui, Liu Mei
Department of Rheumatology and Immunology, Jining No. 1 People's Hospital, Jining, China.
Department of Gastroenterology, Jining No. 1 People's Hospital, Jining, China.
Cell Physiol Biochem. 2018;49(6):2229-2239. doi: 10.1159/000493826. Epub 2018 Sep 26.
BACKGROUND/AIMS: Inflammatory skin diseases are the most common problems in dermatology. Schizandrin A (SchA) has been reported to have anti-inflammatory properties. Herein, we aimed to investigate the protective effects of SchA on lipopolysaccharide (LPS)-induced injury in keratinocyte HaCaT cells.
Inflammation injury in HaCaT cells was induced by LPS treatment. Cell viability, apoptotic cell rate, and apoptosis-related proteins were analyzed by cell counting kit-8 (CCK-8) assay, Annexin V-(fluorescein isothiocyanate (FITC)/ Propidium Iodide (PI) double staining method, and western blot, respectively. The pro-inflammatory factors were analyzed by western blot and quantified by enzyme linked immunosorbent assay (ELISA). Expression of miR-127 in SchA-treated cells was analyzed by qRT-PCR. The effects of SchA on activations of p38MAPK/ERK and JNK pathways were analyzed by western blot.
SchA protected HaCaT cells from LPS-induced inflammation damage via promoting cell viability, suppressing apoptosis. Meanwhile, SchA inhibited IL-1β, IL-6, and TNF-α expression. miR-127 expression was up-regulated in LPS-treated HaCaT cells but down-regulated after SchA treatment. Overexpression of miR-127 inhibited cell growth and induced expression of IL-1β, IL-6 and TNF-α. Additionally, miR-127 overexpression impaired the protective effects of SchA, implying miR-127 might be correlated to the anti-inflammation property of SchA and also involved in inactivation of p38MAPK/ERK and JNK pathways by SchA.
miR-127 is involved in the protective functions of SchA on LPS-induced inflammation injury in human keratinocyte cell HaCaT, which might inactivates of p38MAPK/ERK and JNK signaling pathways in HaCaT cells.
背景/目的：炎症性皮肤病是皮肤科最常见的问题。据报道，五味子甲素（SchA）具有抗炎特性。在此，我们旨在研究SchA对脂多糖（LPS）诱导的角质形成细胞HaCaT细胞损伤的保护作用。
通过LPS处理诱导HaCaT细胞发生炎症损伤。分别采用细胞计数试剂盒-8（CCK-8）法、膜联蛋白V-异硫氰酸荧光素（FITC）/碘化丙啶（PI）双染法和蛋白质免疫印迹法分析细胞活力、凋亡细胞率及凋亡相关蛋白。通过蛋白质免疫印迹法分析促炎因子，并采用酶联免疫吸附测定（ELISA）进行定量。通过qRT-PCR分析SchA处理细胞中miR-127的表达。通过蛋白质免疫印迹法分析SchA对p38丝裂原活化蛋白激酶/细胞外信号调节激酶（p38MAPK/ERK）和c-Jun氨基末端激酶（JNK）通路激活的影响。
SchA通过促进细胞活力、抑制凋亡保护HaCaT细胞免受LPS诱导的炎症损伤。同时，SchA抑制白细胞介素-1β（IL-1β）、IL-6和肿瘤坏死因子-α（TNF-α）的表达。miR-127在LPS处理的HaCaT细胞中表达上调，但在SchA处理后下调。miR-127过表达抑制细胞生长并诱导IL-1β、IL-6和TNF-α的表达。此外，miR-127过表达削弱了SchA的保护作用，这意味着miR-127可能与SchA的抗炎特性相关，并且也参与SchA对p38MAPK/ERK和JNK通路的失活作用。
miR-127参与SchA对LPS诱导的人角质形成细胞HaCaT炎症损伤的保护作用，这可能使HaCaT细胞中的p38MAPK/ERK和JNK信号通路失活。
