青藤碱对佐剂性关节炎大鼠巨噬细胞和滑膜细胞细胞因子表达的影响。

Effect of sinomenine on cytokine expression of macrophages and synoviocytes in adjuvant arthritis rats.

作者信息

Wang Yong, Fang Yongfei, Huang Wenhua, Zhou Xin, Wang Minghai, Zhong Bing, Peng Daizhi

机构信息

Department of Traditional Chinese Medicine, Southwest Hospital, Third Military Medical University, Chongqing 400038, PR China.

出版信息

J Ethnopharmacol. 2005 Apr 8;98(1-2):37-43. doi: 10.1016/j.jep.2004.12.022.

DOI:10.1016/j.jep.2004.12.022

PMID:15763362

Abstract

For exploring the mechanism of the anti-inflammatory and anti-rheumatic effect of sinomenine (SN), the actions of different dosage of SN were observed in vitro on the expression of cytokines, tumor necrosis factor (TNF-alpha) and interleukin-1beta (IL-1beta), as well as the activity of nuclear factor-kappaB (NF-kappaB) and the inhibitory kappaB-alpha (IkappaB-alpha) protein level of peritoneal macrophages (PMs) and synoviocytes in adjuvant arthritis (AA) rats. In this study, the experimental rat model of AA was used and PMs and synoviocytes were collected. The mRNAs of TNF-alpha and IL-1beta were detected with reverse transcription-polymerase chain reaction (RT-PCR) and NF-kappaB activity was measured by electrophoretic mobility shift assay (EMSA). The IkappaB-alpha protein level in the cytoplasma was detected by Western blot. Our results showed that expression of mRNAs of TNF-alpha and IL-1beta and NF-kappaB activity by PMs and synoviocytes were markedly increased compared to control group (P<0.05). In a definite concentration ranging from 30 to 120 microg/ml, SN showed inhibiting effect on the NF-kappaB activity and the expression of the mRNAs of TNF-alpha and IL-1beta in AA rats in a concentration-dependent manner (P<0.05). Positive correlations were found between changes of NF-kappaB activity and expression of TNF-alpha mRNA and IL-1beta mRNA (P<0.01). IkappaB-alpha protein level was increased by various dosages of SN in comparison with control group (P<0.01). In conclusion, SN decreases the mRNA expression of TNF-alpha and IL-1beta by inhibiting the NF-kappaB binding activity, which is mediated through up-regulating the IkappaB-alpha expression of PMs and synoviocytes in AA rats.

摘要

为探讨青藤碱（SN）抗炎抗风湿作用的机制，观察了不同剂量SN体外对佐剂性关节炎（AA）大鼠腹腔巨噬细胞（PMs）和滑膜细胞中细胞因子、肿瘤坏死因子（TNF-α）和白细胞介素-1β（IL-1β）表达，以及核因子-κB（NF-κB）活性和抑制性κB-α（IkappaB-α）蛋白水平的影响。本研究采用AA实验大鼠模型，收集PMs和滑膜细胞。用逆转录-聚合酶链反应（RT-PCR）检测TNF-α和IL-1β的mRNA，用电泳迁移率变动分析（EMSA）测定NF-κB活性。用蛋白质免疫印迹法检测细胞质中IkappaB-α蛋白水平。结果显示，与对照组相比，PMs和滑膜细胞中TNF-α和IL-1β的mRNA表达及NF-κB活性显著增加（P<0.05）。在30至120μg/ml的一定浓度范围内，SN对AA大鼠的NF-κB活性及TNF-α和IL-1β的mRNA表达呈浓度依赖性抑制作用（P<0.05）。NF-κB活性变化与TNF-α mRNA和IL-1β mRNA表达之间呈正相关（P<0.01）。与对照组相比，不同剂量的SN均可使IkappaB-α蛋白水平升高（P<0.01）。综上所述，SN通过抑制NF-κB结合活性降低TNF-α和IL-1β的mRNA表达，这一过程是通过上调AA大鼠PMs和滑膜细胞的IkappaB-α表达介导的。