精氨酸-甘氨酸-天冬氨酸对氧化苦参碱脂质体减轻肝纤维化的增强作用研究

摘要

Objective To investigate whether Arginine-Glycin-Aspartic acid (RGD) could enhance Oxymatrine liposomes (OXYL) theraputic effect on CCl4-induced hepatic fibrosis in rat. Methods CCl4 -induced hepatic fibrosis model was constructed based on rats. Different formulations of oxymatrine were established in this study, i. e. RGD-OXYL and OXYL. Animals were diveded into five groups, which was control group, CCl4 -induced hepatic fibrosis group, OXYL group, RGD-OXYL group and RGD-Liposomes control group. To evaluate the anti-fibrotic effect, levels of alkaline phosphatase, hepatic histopathology (HE and Masson staining) were detected. Moreover,fibrosis-related gene expression of matrixmetallopeptidase-2 (MMP-2), tissue inhibitor of metalloproteinases-1 ( TIMP-1) were determined via semi-quantitative polymerase chain reaction. Results Oxymatrine can attenuate CCl4 -induced hepatic fibrosis, as defined by reducing serum alkaline phosphatase (344. 47±27. 52 vs 550. 69±43. 78, P<0.05), attenuating liver injury and improving collagen deposits (2. 36%±0. 09% vs 7. 70%±0. 60%, P<0. 05) and down regulating fibrosis-related gene expression, i. e. , MMP-2, TIMP-1 (P <0.05). RGD could enhance the therapeutic effect of OM in terms of serum alkaline phosphatase (272. 51±19. 55 vs 344.47±27. 52, P< 0. 05), liver injury, collagen deposits (0. 26%±0. 09% vs 2. 36%±0.09%, P<0.05) and down regulating fibrosis-related gene expression, i. e. , MMP-2, TIMP-1 ( P < 0.05).Conclusion OXYL could attenuate CCL4-induced hepatic fibrosis and inhibit fibrosis-related gene expression. RGD could enhance theraputic effect. of OXYL.%目的 探讨精氨酸-甘氨酸-天冬氨酸(Arginine-Glycin-Aspartic acid,RGD)三肽序列对氧化苦参碱脂质体(oxymatrine liposomes,OXYL)治疗四氯化碳诱导的肝纤维化 ( hepatic fibrosis,HF)作用的影响.方法 建立四氯化碳诱导的大鼠HF模型,制备RGD-OXYL和OXYL.动物分组为:正常对照组;肝纤维化模型组;OXYL治疗组;RGD-OXYL治疗组;RGD脂质体组.检测各组大鼠血清ALP,利用HE染色和Masson染色评价肝脏病理损伤及细胞外基质沉积情况.分离各组大鼠的肝脏组织,利用半定量PCR检测纤维化相关基因(TIMP-1,MMP-2)表达.结果 成功合成OXYL及RGD-OXYL.与肝纤维化组比较,OXYL治疗组大鼠的ALP水平下降(344.47±27.52 vs 550.69±43.78,P<0.05)、肝损伤减轻、细胞外基质沉积面积显著减少(2.36%±0.09% vs 7.70%±0.60%,P<0.05)、纤维化相关基因(TIMP-1,MMP-2)表达下调;与OXYL治疗组大鼠相比,RGD-OXYL治疗组大鼠ALP水平(272.51±19.55 vs 344.47±27.52,P<0.05)和肝损伤及细胞外基质沉积面积(0.26%±0.09% vs 2.36%±0.09%,P<0.05)及纤维化相关基因(TIMP-1,MMP-2)表达等指标均有均有进一步改善.结论 氧化苦参碱脂质体可减轻四氯化碳诱导的肝纤维化,抑制纤维化相关基因表达.RGD偶联的OXYL治疗效果优于OXYL.