吡格列酮对大鼠心脏成纤维细胞胶原合成的研究

Effects of Pioglitazone on Collagen Synthesis in Cardiac Fibroblasts of Rat.

【目的】研究胰岛素增敏剂盐酸吡格列酮(PIO)对培养的Wistar大鼠心脏成纤维细胞(CFs)胶原合成的作用,并进一步探讨其与一氧化氮-氧化氮合酶(NO-NOS)活性的关系。【方法】采用胶原酶消化法培养Wistar大鼠的CFs,羟脯氨酸比色法测定CFs培养上清胶原含量,硝酸还原酶法测定CFs培养上清NO浓度,分光光度计法测定CFs培养上清NOS活性。【结果】①1×10-7、1×10-6、5×10-6、1×10-5mol/L的PIO作用24h后,CFs培养上清胶原含量分别为:1.87±0.14、1.76±0.06、1.44±0.05、1.37±0.09,除1×10-7mol/L组的PIO外,余各药物干预组与对照组(0.21±0.02)相比,差异非常显著(P<0.01)。②5×10-6mol/L的PIO作用24、48、72h后,药物干预组CFs上清胶原含量分别为:1.54±0.08、1.44±0.05、1.39±0.12,与同一时间点的对照组(1.84±0.04、1.88±0.16、2.41±0.01)相比,亦有非常显著性差异(P<0.01)。③5×10-6mol/L的PIO作用48h后,CFs培养上清NO浓度[(221.7±35.3)μmol/L]和对照组[(112.1±8.9)μmol/L]相比,差异非常显著(P<0.001);NOS活性[(15.38±1.82)U/mL]和对照组[(11.24±0.49)U/mL]相比,亦有非常显著性差异(P<0.001)。【结论】PIO能够抑制大鼠CFs的胶原合成,并具有浓度和时间依赖性,其效应可能与上调NO-NOS活性有关。

[Objective]To evaluate the effects of pioglitazone hydrochloride on collagen synthesis in cardiac fibroblasts（CFs） isolated from Wistar rat and the activity of NO-NOS. [Methods]Isolated and cultured CFs were used as experiment model. Collagen content of CFs culture supernatant was estimated by hydroxyproline chromatography. Nitrate enzyme reverting method was also adopted to evaluate NO content. NOS activity was estimated by spectrophotometer. [Results]①Collagen content of CFs culture supernatant（0. 21±0. 01,0.19±0.01,0. 18±0.01,0.17±0.01） was decreased by pioglitazone in a concentration-dependent manner except 1× 10^-7mol/L group. ②Collagen content of CFs culture supernatant （1.54±0.08,1.44±0.05,1.39±0.12） by 5 × 10^-6PIO in 24 h,48 h,72 h group were significantly lower than those of control group（1.84±0.04,1.88± 0.16,2. 41±0. 01, P〈0.01）. ③NO content and NOS activity of pioglitazone group in CFs culture supernatent E（221.7±35.3） μmol/L, （15.38±1.82） U/mL, respectively）] were significantly higher than those of control group[ （112.1±8.9）μmol/L,（11. 24±0.49 ）U/mL , respectively,（ P 〈0. 001）]. [Conclusion]Insulinsensitizer Pioglitazone Hydrochloride could inhibit collagen content of cardiac fibroblasts in a concentration-dependent and time-dependent manner in vitro, Up-regulating the activity of NOS may take part in signal pathway.