摘要
目的:观察金雀异黄素(genistein,Gen)对Ⅱ型胶原诱导型关节炎(collagen-induced arthritis,CIA)大鼠膝关节成纤维样滑膜细胞(fibroblast-like synoviocyte,FLS)分泌炎症因子白细胞介素1β(interleukin-1β,IL-1β)和肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)的影响。方法:采用Ⅱ型胶原注射建立CIA动物模型,用关节炎指数(arthritis index,AI)分析法评估CIA模型是否建立成功。胶原酶消化法分离培养原代FLS,用流式法检测滑膜细胞血管细胞黏附分子-1(vascular cell ad-hension molecule-1,VCAM-1)的表达。在FLS培养液中加入不同浓度的Gen,用酶联免疫吸附测定法检测FLS培养上清中IL-1β和TNF-α的含量。结果:Ⅱ型胶原致敏后3d,大鼠开始出现关节肿胀,AI逐渐增高,至造模后第3周最为明显,与空白对照组比较差异有统计学意义。大鼠滑膜细胞培养至第4代,检测其VCAM-1的表达高达85.5%,提示所培养的细胞为FLS,即B型滑膜细胞。不同浓度的Gen(100、200、400μmol/L)作用于FLS细胞后,细胞培养上清中TNF-α和IL-1β的分泌不同程度地受到抑制,且抑制作用呈剂量依赖性。结论:金雀异黄素能抑制FLS分泌TNF-α和IL-1β,且抑制作用呈剂量依赖性,这可能是其抗CIA大鼠滑膜炎的作用机制之一。
Objective. To investigate the effects of genistein （Gen） on interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） secreted by fibroblast-like synoviocytes （FLSs） of rats with type Ⅱ collagen-reduced arthritis （CIA）.
Methods： Type Ⅱ collagen was injected to induce arthritis in rats. and arthritis index was applied to evaluate whether the arthritis was induced successfully. The primary FLSs were separated from synovial membranes by using type Ⅱ collagenase digestion. Then the expression of vascular cell adhesion molecule- 1 （VCAM-1） was estimated by flow cytometry （FCM）. After addition of different concentrations of Gen into the FLSs, IL-1β and TNF-α contents in the supernatants were measured by enzyme-linked Jmmunosorbent assay.Results： Three days after collagen injection, the rats started to develop arthrocele and the arthritis index increased gradually. The arthritis index of CIA group was significantly higher than that of the control group. The expression of VCAM-1 was up to 85. 5% in the 4th generation FLSs, indicating that most part of the cultured cells were type-B synoviocytes. After administration of different concentrations of Gen （100, 200, 400 μmol/L）, the contents of IL-1β and TNF-α in supernatants of FLSs were decreased dose-dependently. Conclusion： Genistein can suppress the secretion of TNF-α and IL-1β in FLSs dose-dependently, which may be one of the mechanisms for genistein in inhibiting the arthromeningitis of CIA rats.
出处
《中西医结合学报》
CAS
2009年第7期636-641,共6页
Journal of Chinese Integrative Medicine
基金
江苏省高校自然科学基础研究项目(No.07KJD320245)
