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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
OBJECTIVE: To investigate the effect of COX 2 inhibitor on the proliferation, viability and extracellular matrix production of fibroblasts, and to study the mechanism of COX 2 inhibitor against hepatic fibrosis.Methods: NIH / 3T3 fibroblasts were used as hepatic stellate (HSC) as a substitute model and routinely cultured.COX 2 specific inhibitor celecoxib (final concentration of 5, 10, 20, 40 μmol / L) was added to the cells and detected by MTT assay The effect of celecoxib on the proliferation and viability of NIH / 3T3 cells was detected by radioimmunoassay. The production of extracellular matrix (HA, LN, PCIII, IVC) was detected by radioimmunoassay. COX 2 protein expression.Results: NIH / 3T3 cells COX2 protein positive, is the target of celecoxib; celecoxib at the concentrations used in the cells without significant toxicity, but significantly inhibited cell proliferation in the concentration (P <0.01), inhibiting the synthesis of HA and LN. The inhibitory rates of 5 ~ 40 μmol / L on LN were 1 9.74%, 1 8.16%, 22.0% and 24% .6 5%. The inhibitory rates of HA to 20 ~ 40 μmol / L were 1 7.53% and 24.4%, respectively. The contents of HA and LN were significantly different from those of the control group Significantly (P <0 .0 5) Conclusion: Celecoxib significantly inhibited the outer NIH / 3T3 cell proliferation and the synthesis of HA and LN has certain liver fibrosis in vitro