雷公藤对类风湿关节炎病人软骨细胞一氧化氮合成、一氧化氮合酶活性及其mRNA表达的体外抑制作用  

In vitro inhibitive effects of Tripterygium wilforii on NO production, iNOS activity, and iNOS-mRNA expression in chondrocyrtes of patients with rheumatoid arthritis

作　　者：郭万首[1] 马丽[2] 陶学廉[3] 吴东海[2] 

机构地区：[1]中日友好医院骨科,北京100029 [2]中日友好医院风湿免疫科,北京100029 [3]美国德州大学西南医学中心风湿病科实验室

出　　处：《中华医学杂志》2001年第17期1035-1037,共3页National Medical Journal of China

摘　　要：目的　探讨雷公藤治疗类风湿关节炎 (RA)的作用机制。方法　将RA病人膝关节置换术后的离体软骨经酶消化 ,并分离出软骨细胞进行体外培养 ,用白细胞介素 (IL) 1β刺激 ,同时加用不同浓度的雷公藤、地塞米松及消炎痛培养过夜 ,收集细胞培养上清 ,用Griess试剂测定NO含量。收集软骨细胞 ,利用L 14 C 精氨酸向L 14 C 瓜氨酸的转换 ,测定iNOS活性。提取细胞总RNA ,采用Northernblot方法分析iNOS mRNA的表达。结果　不同浓度的雷公藤 (2、4、8及 16mg/L)对关节软骨细胞NO的合成和iNOS活性均有抑制作用 ,对NO合成的抑制率分别为 10 8%、2 5 4 8%、5 5 17%及 80 4 5 % ,对iNOS活性的抑制率分别为 12 2 9%、2 7 6 7%、5 9 0 4 %及 85 0 6 % ,而且抑制程度与药物剂量成正相关 ,相关系数r为 0 976和 0 974 ,雷公藤对iNOS mRNA的表达也有相应的抑制作用。与之相比地塞米松有明显的抑制作用 ,但无剂量相关 ,而消炎痛只有微弱抑制作用。结论　雷公藤对人关节软骨细胞NO合成的抑制是通过限制iNOS基因转录而完成 。Objective To study the mechanism of treatment of rheumatoid arthritits with Tripterygium wilfordii Hook. f. (TWHF). Methods Chondrocytes from the resected knee joint cartilage of patients with rheumatoid and TWHF, dexamethasone, or(arthritis were isolatred and cultured. IL 1 indomethacin of different concentrations were added into the culture solution overnight. Griess reagent was added to the supernatant to detect the content of NO. Chondrocytes werte collected to examine the iNOS activity by detecting the conversion of L 14C arginine into L 14C citruline. The total RNA of hondrocyte was extracted and the iNOS mRNA was examined by Northen blotting. Results The inhibitory rates of NO production by TWHF of concentrations of 2 mg/L, 4 mg/L, 8 mg/L, and 16 mg/L were 10.8%, 25.48%, 55.17%, and 80.45% respectively. The inhibitory rates of iNOS activity by TWHF of concentrations of 2 mg/L, 4 mg/L, 8 mg/L, and 16 mg/L were 12.29%, 27.67%, 59.04%, and 85.06% respectively. Such inhibitory effects were dosage dependent. TWHF ffectively inhibited the expression of iNOS mRNA induced by IL 1 in chondrocyte ( r =0.976 and 0.974). Dexamethasone inhibited NO production, iNOS activity, and expression of iNOS mRNA significantly but not dosage dependently. Indomethcin only had weak inhibitory effect. Conclusion TWHF inhibits NO production in chondrocytes by limiting the transcription of iNOS gene, which may be one of the mechanisms of treatment of RA with TWHF.

关 键 词：一氧化氮 一氧化氮合酶 类风湿性关节炎 雷公藤 治疗 mRNA 

分 类 号：R593.22[医药卫生—内科学]