葡萄籽原花青素预防大鼠肝纤维化的实验研究

目的观察葡萄籽原花青素(GSPE)抑制四氯化碳(CCL4)诱导大鼠肝纤维化形成的效果,并探讨其可能的作用机制。方法 50只大鼠随机选取20只分为空白对照组(10只)和溶剂组(10只),其他大鼠采用CCL4腹腔注射法,每周2次,8周制造大鼠肝纤维化模型,将造模成功的24只大鼠分为模型组(12只)和GSPE干预组(12只)。GSPE干预组从第5周给予GSPE 150 mg/(kg.d)灌胃,8周后处死大鼠,留取肝脏和血清,采用自动生化分析仪检测血清中丙氨酸氨基转移酶(ALT)、天门冬酸氨基转移酶(AST)、总胆红素(TBIL),放射免疫法检测血清透明质酸(HA)、层黏连蛋白(LN)、Ⅲ型前胶原(PCⅢ),肝组织碱水解法检测羟脯氨酸(HyP),化学比色法检测肝组织超氧化物歧化酶(SOD)、丙二醛(MDA)、还原型谷胱甘肽(GSH),同时取固定部位肝脏组织HE染色,观察其肝纤维化程度。结果与空白对照组比较,模型组大鼠血清中的ALT、AST、TBIL、HA、LN、PCⅢ水平均显著升高(P<0.05),肝组织中GSH和SOD降低,MDA和Hyp含量升高(P<0.05);与模型组比较,GSPE干预组大鼠血清ALT、AST、TBIL、HA、LN、PCⅢ水平均显著降低(P<0.05),肝组织中GSH和SOD升高,MDA和Hyp含量降低(P<0.05);与模型组比较,GSPE干预组肝组织纤维化病变明显减轻,差异有统计学意义(P<0.05)。结论GSPE对CCL4诱导的大鼠肝纤维化形成具有明显的抑制作用,其作用机制可能与保护肝细胞、抑制脂质过氧化和抑制胶原合成有关。

Grape seed proanthocyanidin extract in the prevention of rat liver fibrosis

Objective To observe the therapeutic effect of grape seed proanthocyanidin extract(GSPE) on Carbon tetrachloride-induced hepatic fibrosis in rats and explore its therapeutic mechanism.Methods 20 of 50 rats were randomly selected as the control group(n=10) and the solvent group(n=10).The remaining 30 rats were regarded as models of hepatic fibrosis which were established by intraperitoneal injection of carbon tetrachloride twice a week for 8 weeks.The 24 successful model rats were divided into the model group(n=12) and the GSPE group(n=12),with GSPE 150 mg/(kg·d) being given to the GSPE group rats from 5th week.After eight weeks,all rats were executed,and the liver and serum were stored for further investigation.Serum alanine aminotransferase(ALT)、aspartate aminotransferase(AST) and total bilirubin(TBIL) were tested by automatic biochemical analyzer.Hexadecenoic acid(HA) 、 laminin(LN) and Procollagen Ⅲ(PCⅢ) were tested by radioimmunoassay.Hydroxyproline(HyP) was tested by liver tissue alkaline hydrolysis.Superoxide dismutase(SOD) 、malondialdehyde(MDA) and reduced glutathione hormone(GSH) were determined by chemical colorimetric method.Additionally,liver samples from fixed part were stained by HE to trace the degree of fibrosis.Results Levels of serum ALT、AST、TBIL、HA、LN、PCⅢ in the rats of the model group were significantly raised(P<0.05) compared to those of the control group.Activities of GSH and SOD in hepatic tissue decreased and levels of MDA and Hyp obviously increased(P<0.05).Levels of serum ALT、AST 、TBIL、HA、LN、PCⅢ in the GSPE group were significantly reduced(P<0.05) in comparison to those of the model group.Activities of GSH and SOD in hepatic tissue increased and levels of MDA and Hyp decreased(P<0.05).In addition,the GSPE group had significantly improved histological changes of hepatic fibrosis(P<0.05).Conclusion GSPE plays an inhibitory role in CCL4-induced hepatic fibrosis in rats,probably through protection of hepatic cell,alleviation of injury induced by lipid peroxidation and inhibition on collagen fiber synthesis.