摘要
目的:探讨白介素17(Interleukin-17,IL17)对类风湿关节炎病人外周血淋巴细胞表面(Lymphatic cell)核因子—kB受体活化因子配体(receptor activator of nuclear factor–kB ligand, RANKL )、骨骼保护因子(osteoprotegerin ,OPG)mRNA表达的影响;阐述IL17是否通过上调淋巴细胞表面RANKL因子的表达来增加破骨细胞数量和活性,从而引起类风湿关节炎病人的骨质破坏。方法:取类风湿关节炎病人的外周血分离培养其淋巴细胞,并用免疫组化的方法对其进行鉴定,用不同浓度的IL17(10-9 ug/ml、10-8 ug/ml、10-7 ug/ml、10-6ug/ml、10-5ug/ml)分别处理分离培养的淋巴细胞,24小时后使用Trizol抽提总的RNA,通过半定量逆转录聚合酶链反应(RT-PCR)来观察不同浓度IL17对类风湿关节炎病人外周血淋巴细胞表面RANKL、OPG mRNA表达的影响及RANKL/OPG的比率变化,并将10-6ug/ml浓度组设定时间(0、4、16、24、48小时),在不同的时间分别抽提总的RNA用上述同样的方法测定不同时间IL17对类风湿关节炎患者外周血淋巴细胞表面RANKL、OPGmRNA表达量的影响及RANKL/OPG的比值变化。结果:10-9—10-5ug/ml IL17在24小时后可上调类风湿关节炎病人淋巴细胞表面RANKL mRNA的表达并呈剂量依赖型,但抑制OPG mRNA的表达,且RANKL/OPG比率呈上升趋势(P<0.05)。随着白介素17(浓度10-6ug/ml)作用时间的延长,淋巴细胞表面RANKL mRNA的上调作用、OPG mRNA的抑制作用逐渐增强,RANKL/OPG比率呈明显上升趋势.不同时间段白介素17对RANKL、OPG mRNA表达量的变化,RANKL/OPG比率变化与正常对照组相比有显著差异(P〈0.01)结论:IL17有可能通过调节类风湿关节炎病人淋巴细胞表面RANKL、OPG mRNA量的表达和RANKL/OPG mRNA比例变化,来间接地调节破骨细胞的分化和活性,引起骨质破坏。
To study the expression of receptor activator of nuclear factor-kB ligand(RANKL) and osteoprotegerin(OPG) on the peripheral blood lymphatic cell of the patient with rheumatoid arthritis;To explore whether IL17 increase the number and activity of osteoclast by upregulating the expression of RANKL factor and causing bone destruction of rheumatoid arthritis patients. METHODS: Isolating and culturing lymphatic cell from peripheral blood of patient with rheumatoid arthritis ,and to identify lymphatic cell by immunohistochemical methods. Isolated lymphatic cell is treated by IL17 of different concentration (10-9ug/ml ,10-8ug/ml,10-7ug/ml,10-6ug/ml,10-5ug/ml). After incubation for 24h , semi-quantitative RT-PCR was performd to detect the expression of OPG,RANKL mRNA and the ratio of RANKL/OPG usingβ-actin mRNA level as the internal control.And the 10-6ug / ml concentration group setting time (0,4,16,24,48 hours) . At different times, the total RNA was extracted using the same method to detect the expression of RANKL, OPG mRNA and the ratio of RANKL/OPG. RESULTS:After 24 hours 10-9-10-6mol/L IL17 up– regulated the expression of RANKL mRNA with dose-dependent ,but inhibited the expression of OPG mRNA. Ratio of RANKL /OPG mRNA in human lymphatic cell showed increasing tendency(P<0.05). As the extention of time,IL17 upregulated RANKL mRNA and inhibited the expression of OPG mRNA is increasingly heighten, but RANKL/OPG ratio was significantly increased. Compared with the control group, in the periods of different times changes of RANKLOPG mRNA and RANKL/OPG ratio was significantly different.(p<0.01)CONCLUSION: IL17 may regulate the differentiation and activity of osteoclasts by influencing RANKL, OPG mRNA and the ratio of RANKL/OPG mRNA,and thus indirectly regulate osteoclast differentiation and activity, causing bone destruction.
引文
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