秋水仙碱对成纤维细胞产生细胞因子以及分泌细胞外基质的影响  

Effects of colchicine on synthesis and excretion of cytokines and extracellular matrix by human renal fibroblasts

作　　者：黄文彦[1] 孙骅[2] 潘晓勤[2] 费莉[2] 郭梅[2] 鲍华英[2] 陈荣华[2] 姜新猷[1] 

机构地区：[1]南京医科大学第二附属医院儿科 [2]南京医科大学小儿肾脏病研究中心

出　　处：《中华儿科杂志》2004年第7期524-528,共5页Chinese Journal of Pediatrics

基　　金：国家自然科学基金资助项目 ( 3 0 2 70 619);江苏省卫生厅重点资助项目(H990 6)

摘　　要：目的　阐明秋水仙碱对体外培养人肾脏成纤维细胞 (FB)产生炎症因子转化生长因子 β1(TGF β1)、白细胞介素 1β(IL 1β)以及细胞外基质的影响。 方法　体外培养人胚胎肾脏FB ,经不同浓度秋水仙碱 ( 5 0 μmol/L、10 0 μmol/L、2 0 0 μmol/L、4 0 0 μmol/L)预处理 1h后 ,加入含 10 0μg/ml脂多糖 (LPS)的FB完全培养液继续培养 18h ,以未经LPS刺激也无秋水仙碱预处理的FB细胞为对照。实验终点收集细胞和上清。分别利用RT PCR方法观察秋水仙碱对FB产生TGF β1和IL 1βmRNA表达的影响 ;ELISA法测定细胞上清TGF β1、IL 1β以及胶原COLⅢ、COLⅣ蛋白含量。结果　 ( 1)单纯 10 0 μg/mlLPS刺激后 ,FB产生TGF β1和IL 1βmRNA分别较未经刺激的正常对照组上调 3倍 (RI值分别为 6 6 1± 1 6、2 2 3± 2 0 ,q =5 90 5 ,P =0 0 0 2 )和 4 7倍 (RI值分别为2 2 0± 2 2、4 7± 0 8,q =10 6 8,P =0 0 0 9)。细胞上清TGF β1和IL 1β蛋白含量均明显高于基础含量 [TGF β1分别为 :( 5 16± 14 )pg/ml和 ( 4 2 0± 5 ) pg/ml(q =80 3,P =0 0 12 ) ,IL 1β分别为 :( 3 4± 0 3) pg/ml和 ( 0 3± 0 1) pg/ml( q=2 97 9,P =0 0 0 3) ]。 ( 2 )秋水仙碱明显抑制FB之TGF β1mRNA以及蛋白表达 。Objective Renal interstitial fibrosis is the final common pathway leading to end stage renal failure for progressive renal disease of various types. The present study was undertaken to add to the knowledge on colchicine′s antiinflammatory and antifibrotic properties confirmed by both human and experimental studies. As the main effector cells, fibroblasts have a central role in the pathogenesis of renal fibrosis. This study aimed to investigate the effects of colchicine on the synthesis and excretion of cytokines transforming growth factor β1 (TGF β1), interleukin 1β (IL 1β) and extracellular matrix (collagen Ⅲ, collagen Ⅳ) in human renal fibroblast. Methods Various concentrations of colchicine (5 0 μmol/L, 10 0 μmol/L,20 0 μmol/L, 40 0 μmol/L) were used to pretreat human embryo renal fibroblasts for 1 hour which were cultured in vitro and then stimulated by 10 0 μg/ml of lipopolysaccharide (LPS). After 18 hours, these fibroblasts and their supernatant were collected. The expression of TGF β1 mRNA, IL 1β mRNA in the cells was studied by using RT PCR, and the excretion of TGF β1, IL 1β, collagen Ⅲ and collagen IV by the fibroblasts was assessed by ELISA respectively. Results (1) By pure stimulation with 10 0 μg/ml LPS,the expression of TGF β1 mRNA and IL 1β mRNA of fibroblasts was up regulated 3 times (66 1±1 6 vs 22 3±2 0, q =590 5, P =0 002) and 4 7 times (22 0±2 2 vs 4 7±0 8, q =106 8, P =0 009), respectively. The protein excretion of TGF β1 and IL 1β was remarkably increased as well compared with the control group [TGF β1: (516±14)pg/ml vs (420±5)pg/ml ( q =80 3, P =0 012), IL 1β:(3 4±0 3)pg/ml vs. (0 3±0 1)pg/ml ( q =297 9, P =0 003)]. (2) Colchicine could inhibit the expression of TGF β1 mRNA and protein in a dose dependent manner. IL 1β mRNA and protein were both up regulated by colchicine. (3) LPS could not stimulate the excretion of extracellular matrix by fibroblasts, but

关 键 词：秋水仙碱 成纤维细胞 细胞因子 细胞外基质 转化生长因子-β1 白细胞介素-1Β 肾间质纤维化 

分 类 号：R965[医药卫生—药理学]