Zhou Qi, Liu Shumin, Yua Donghua, Zhang Ning
Research Institute of Chinese Medicine, Harbin 150040, PR China.
Technological Innovation Team of Basic Theory Study Research of Institution of Higher Education in Heilongjiang Province, Heilongjiang University of Chinese Medicine, Harbin 150040, PR China.
Pharmacogn Mag. 2016 Jul-Sep;12(47):235-40. doi: 10.4103/0973-1296.186344.
Dioscorea nipponica Makino is one of the most common used traditional Chinese drugs which are used to treat gouty arthritis (GA). Nuclear factor-κB (NF-κB) pathway plays an important role during this process. In the present study, we investigated the effects of total saponins from D. nipponica Makino (TDN) on NF-κB pathway in interleukin-1β (IL-1β) induced fibroblast-like synoviocytes (FLS).
FLS were divided into three groups: Normal group, model group, which was given 10 μg/L IL-1β to induce the proliferation, and TDN group (10 μg/L IL-1β +100 μg/L TDN). 1 h, 24 h, 48 h, and 72 h after treating, immune fluorescence method was used to detect the cell location of NF-κB p65. Electrophoretic mobility shift assay was used to detect the activation of NF-κB p65. Western blot method was used to detect the protein expressions of NF-κB p65, IκBα, and p-IκBα.
TDN could inhibit the activation and transfer of NF-κB p65. As time went on, the expression of NF-κB p65 in the cytoplasm was decreased while it was increased in the nucleus. The expression of p-IκBα was increased, whereas the expression of IκBα was not changed. TDN could regulate these abnormal expressions.
TDN may treat GA by regulating NF-κB signal pathway.
TDN could inhibit the transfer of NF-κB p65.TDN could inhibit the activation of NF-κB p65.TDN could inhibit the expression of p-IκBa. Abbreviations used: TDN: Total saponins from Dioscorea nipponica Makino, GA: Gouty arthritis, FLS: Fibroblast-like synoviocytes, IL-1β: Interleukin-1 beta, IF: Immune fluorescence, EMSA: Electrophoretic mobility shift assay, WB: Western blot.
穿山龙是治疗痛风性关节炎(GA)最常用的传统中药之一。核因子-κB(NF-κB)通路在此过程中起重要作用。在本研究中,我们研究了穿山龙总皂苷(TDN)对白细胞介素-1β(IL-1β)诱导的成纤维样滑膜细胞(FLS)中NF-κB通路的影响。
FLS分为三组:正常组、模型组(给予10μg/L IL-1β诱导增殖)和TDN组(10μg/L IL-1β +100μg/L TDN)。处理后1小时、24小时、48小时和72小时,采用免疫荧光法检测NF-κB p65的细胞定位。采用电泳迁移率变动分析检测NF-κB p65的活化情况。采用蛋白质免疫印迹法检测NF-κB p65、IκBα和p-IκBα的蛋白表达。
TDN可抑制NF-κB p65的活化和转位。随着时间的推移,NF-κB p65在细胞质中的表达降低,而在细胞核中的表达增加。p-IκBα的表达增加,而IκBα的表达未改变。TDN可调节这些异常表达。
TDN可能通过调节NF-κB信号通路治疗GA。
TDN可抑制NF-κB p65的转位。TDN可抑制NF-κB p65的活化。TDN可抑制p-IκBa的表达。缩写词:TDN:穿山龙总皂苷,GA:痛风性关节炎,FLS:成纤维样滑膜细胞,IL-1β:白细胞介素-1β,IF:免疫荧光,EMSA:电泳迁移率变动分析,WB:蛋白质免疫印迹