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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 ,而
Objective To elucidate the effects of colchicine on the production of transforming growth factor β1 (TGFβ1), interleukin 1β (IL 1β) and extracellular matrix in cultured human renal fibroblasts. Methods Human embryonic kidney FB was cultured in vitro and pretreated with different concentrations of colchicine (50 μmol / L, 100 μmol / L, 200 μmol / L, 400 μmol / L) for 1 h. / ml lipopolysaccharide (LPS) FB complete culture medium to continue training 18h, without LPS stimulation nor colchicine pretreatment of FB cells as a control. The end of the experiment collected cells and supernatant. The effects of colchicine on the expression of TGFβ1 and IL-1β mRNA by FB were observed by RT-PCR. The contents of TGFβ1, IL-1β and collagen COLⅢ and COLⅣ were determined by ELISA. Results (1) After stimulated with LPS alone at 10 0 μg / ml, the TGFβ1 and IL 1β mRNA levels in FBs were upregulated 3-fold (616 ± 1 6,2 2 3 ± 2 0, respectively) compared with the untreated normal controls , q = 5 905, P = 0 0 0 2) and 4 7 times (RI values were 220 ± 2 2,4 7 ± 0 8, q = 10 6 8, P = 0 0 0 9, respectively). The contents of TGFβ1 and IL 1β in the supernatant of the cells were significantly higher than those of the basal [TGFβ1 (5 16 ± 14) pg / ml and (42 ± 5) pg / ml, respectively 0 0 12), IL 1β were: (34 ± 0 3) pg / ml and (0 3 ± 0 1) pg / ml, respectively (q = 2 97 9, P = 0 0 0 3)]. (2) Colchicine significantly inhibited TGFβ1 mRNA and protein expression in FB