【摘要】 目的通过体外培养骨关节炎(OA)软骨细胞体系,了解白藜芦醇对白细胞介素(IL)-lβ诱导OA软骨细胞蛋白聚糖代谢的影响。方法软骨细胞采自OA患者,采用分阶段酶消化法体外原代培养人软骨细胞。在培养液中加入IL-lβ诱导剂,设立兔血清对照组、实验对照组和白藜芦醇加药组;给予实验兔以白藜芦醇剂量灌胃后,抽取含药血清培养细胞。采用3种方法了解蛋白聚糖的代谢水平:1)用二甲基亚甲蓝分光光度法(DMMB法)检测细胞中及培养液中的糖安多糖(GAG)含量;2)酶联免疫吸附法检测培养液中蛋白聚糖的不同代谢片段(Mab-3B3和5D4);3)反转录聚合酶链反应(RT-PCR)半定量法检测软骨细胞的蛋白聚糖、基质金属蛋白酶-1(matrix metalLoproteinases,MMPs-1)和MMPs-3的mRNA表达水平。结果各浓度白藜芦醇含药血清组体外培养的软骨细胞释放入培养液中GAG百分比均值明显低于对照组,两者相比差异有统计学意义(P<0.01),随白藜芦醇的增加,释放入培养液中GAG的百分比逐渐降低;3B3含量的均值较对照组相比明显增高(P<0.01),与白藜芦醇浓度呈正相关,释放入培养液中5D4含量的均值则明显降低(P<0.05),与白藜芦醇浓度呈负相关;白藜芦醇可以增加OA患者软骨细胞和IL-1β诱导的OA患者软骨细胞蛋白聚糖mR-NA表达,减低MMPs-1、MMPs-3 mRNA的表达。结论白藜芦醇可以抑制IL-1β对OA患者软骨细胞蛋白聚糖代谢的促进作用,达到保护软骨,防止OA的目的 。更多还原

【Abstract】 【Objective】 To evaluate the effects of resveratrol induced by interleukin(IL)-1 beta on aggrecan metabolism in cultured human osteoarthritis(OA) chondrocytes,and to culture osteoarthritis chondrocytes in vito.【Methods】 Chondrocytes isolated from knee of OA patients were cultured by phased enzymatic digestion.Sera containing resveratrol of the experimental animals were obtained after orally administrated resveratrol at the dosages that equal to human.Glycosaminoglycan(GAG) content in the chondrocytes and matrix were measured by 1,9-dimethylmethylene blue spectrophotometric assay.Aggrecan catabolic fragments(Mab-3B3and 5D4) in matrix were measured by ELISA.RNA was isolated from chondrocytes and analyzed for the expression of aggrecan,matrix metalloproteinases(MMPs-1),MMPs-3 and 18S as control.【Results】 Average level of GAG in matrix samples of resveratrol-containing groups decreased significantly comparing with the control groups(P <0.01).Similar decrease could be observed in fragments of 5D4 in matrix of resveratrol-containing groups.But there was an increase could be observed in the 3B3 fragments.Expression of aggrecan mRNA in the resveratrol-containing groups was higher than that of the control groups.mRNA of MMPs-1 and mRNA of MMPs-3 of resveratrol-containing groups were lower than that of the control groups.【Conclusion】 Resveratrol can markedly inhibit promoting effect of aggrecan metabolism induced by interleukin-lβ.To protect cartilage and prohibit OA.更多还原