【摘要】 目的 :探讨COX 2抑制剂对成纤维细胞增殖、活力及细胞外基质产生的影响 ,研究COX 2抑制剂抗肝纤维化的机制 .方法 :采用体外培养的NIH/ 3T3成纤维细胞作为肝星状细胞 (HSC)的替代模型 ,常规培养 .COX 2特异性抑制剂塞来昔布 (celecoxib) (终浓度为 5 ,1 0 ,2 0 ,4 0 μmol/L)加入细胞中 ,用MTT法检测塞来昔布对NIH/ 3T3细胞的增殖 ,活力的影响 ;用放射免疫法测定培养上清细胞外基质 (HA ,LN ,PCIII,IVC)的生成 ;用Westernblot法检测NIH/ 3T3细胞上的COX 2的蛋白表达 .结果 :NIH/ 3T3细胞的COX 2蛋白表达阳性 ,是塞来昔布作用靶点 ;塞来昔布在所用浓度下对细胞无明显毒性 ,但可显著抑制细胞增殖 ,呈浓度依赖性 (P <0 .0 1 ) ,抑制HA及LN的合成 .5～ 4 0 μmoL/L对LN的抑制率分别为 1 9.74 % ,1 8.1 6 % ,2 2 .0 5 %和 2 4 .6 5 % ;2 0～ 4 0 μmoL/L对HA的抑制率分别为 1 7.5 3%和 2 4 .2 4 % ,HA及LN的含量与对照组比较差异显著 (P <0 .0 5 ) .结论 :塞来昔布可显著抑制NIH/3T3细胞增殖和细胞外HA及LN的合成 ,在体外具有一定的抗肝纤维化作用更多还原

【Abstract】 AIM: To study the effects of Celecoxib, a specific COX 2 inhibitor, on the proliferation, viability and extracellular matrix production of fibroblasts, and to explore the mechanisms involved in its protection against liver fibrosis. METHODS: NIH/3T3 fibroblasts were cultured in vitro , as the substitutive model of hepatic satellate cells (HSC). The dosage of Celecoxib used was 5, 10, 20 and 40 μmol/L, respectively. The effects of Celecoxib on the cell proliferation was measured by methabenzthiazuron (MTT) assay and the extracellular matrix (HA, LN, PCIII and IVC) in culture medium was detected by radioimmunoassay. The expression of COX 2 in NIH3T3 was analyzed by Western blot. RESULTS: Western blot indicated that COX 2 protein was expressed in NIH3T3 cells, the drug target of Celecoxib. Celecoxib showed no significant toxicity to NIH3T3 cells, but significantly inhibited the cell proliferation in a dose dependent manner ( P <0.01), and the synthesis of HA and LA as well. With 5-40 μmol/L of Celecoxib, the inhibition rate of LA was 19.74%, 18.16%, 22.05% and 24.65% respectively and with 20-40 μmol/L of Celecoxib, the inhibition rate of HA was 17.53 % and 24.24% respectively, which differed significantly from those in the control groups ( P < 0.05). CONCLUSION: Celecoxib can significantly inhibit NIH/3T3 cell proliferation and ECM (HA, LN) production and may play a role in anti liver fibrosis in vitro.更多还原