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

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.

Arginine-Glycin-Aspartic acid could enhance oxymatrine liposomes theraputic effect on hepatic fibrosis

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-fibrotie effect, levels of alkaline phosphatase, hepatic histopathology （HE and Masson staining） were detected. Moreover,fibrosis-related gene expression of matrix metallopeptidase-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 CCIA-induced hepatic fibrosis and inhibit fibrosis-related gene expression. RGD could enhance theraputic effect, of OXYL.