肝再生增强因子对大鼠肝纤维化的治疗作用及机制研究
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摘要
肝纤维化(liver fibrosis)是各种慢性肝病的共同病理过程,严重的肝纤维化可发展成肝硬化甚至肝癌。在我国,肝纤维化、肝硬化多与病毒性肝炎有关,往往是由于慢性病毒性肝炎迁延难愈而造成的。针对慢性肝炎肝纤维化的病理特征,本文采用CCl4和PS诱导的大鼠肝纤维化模型,研究ALR对大鼠肝纤维化的治疗作用并探讨其抗肝纤维化的机制。
研究发现在各种急、慢性肝损伤动物模型中,ALR可促进肝细胞再生,减轻肝损伤,改善肝功能,增加动物存活率和存活时间。本文采用尾静脉注射pcDNA3.1-hALR重组质粒治疗毒物诱导和免疫性大鼠肝纤维化。
研究结果显示,ALR基因治疗可明显减轻大鼠肝损伤和肝纤维化,其机制可能是抑制肝细胞内氧化应激,下调各种致纤维化因子,阻止肝星状细胞的活化。ALR治疗可显著减少α-SMA的表达,减少肝内活性氧簇水平,并下调TGF-β1、PDGF-BB和TIMP-1的表达。与肝硬化模型组大鼠相比,治疗组大鼠的血清AST, ALT和AKP水平明显降低,但是,血清ALB明显高于模型组大鼠而接近健康对照组。血清生化指标的变化与肝组织病理检查结果相符,治疗组大鼠肝组织H&E染色显示肝小叶结构的破坏得到修复,Sirius-red胶原特异性染色显示胶原阳性面积显著减少,qRT-PCR结果表明Ⅰ型胶原和Ⅲ型胶原含量明显减少。与健康对照组比较,模型组大鼠肝脂质过氧化指标MDA含量增加,SOD活性降低。经过ALR治疗后,治疗组大鼠MDA含量降低,SOD活性增加。与健康对照组相比,模型组大鼠肝脏Hyp含量显著增加。而与模型组大鼠比较,治疗组大鼠肝脏Hyp含量明显减少。总之,研究结果提供了新的ALR保护肝脏的机制。
Liver fibrosis is a common pathological process of chronic hepatic disease, which can lead to cirrhosis and increase risk for hepatocellular carcinoma. In China, liver fibrosis and cirrhosis are usually caused by the developing of chronic viral hepatitis. Here we evaluated the therapeutic efficacy of augmenter of liver regeneration (ALR) on liver fibrosis induced by carbon tetrachloride (CC14) and porcine-serum (PS), and investigated potential mechanisms.
Accumulated studies had revealed that ALR appeared to be an important regulator of liver regeneration and had trophic effects on damaged liver in animal models of acute or chronic liver injury. In these studies, exogenous ALR administration ameliorated hepatic damage, enhanced hepatic proliferative capacity post-toxininduced liver injury, or improved survival of toxin-administered animals. The present research was to observe the therapeutic effects of ALR on liver fibrosis and to explore the molecular mechanism of antifibrosis.
ALR might attenuate liver injury and fibrosis by suppressing oxidative stress, down-regulating profibrogenic factors, and blocking HSCs activation. This report demonstrated that ALR therapy diminished α-smooth muscle actin (SMA) expression, decreased intrahepatic reactive oxygen species (ROS) level, and down-regulated transforming growth factor (TGF)-β1, platelet-derived growth factor (PDGF)-BB, and tissue inhibitor of metalloproteinase (TIMP)-1expression. Compared with the model rats, cirrhotic rats treated with ALR showed ameliorated biochemical liver tests. In these animals, serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (AKP) were significantly lower and serum albumin (ALB) significantly higher than in cirrhotic rats and similar to healthy controls. These favorable changes were accompanied by histological improvement with marked reduction of fibrosis and decreased expression of collagen I and IV in ALR-treated rats. Compared with cirrhotic rats, ALR treatment were able to reduce the level of malondialdehyde (MDA) content, increased superoxide dismutase (SOD) activity. Compared healthy control rats, the Hyp content of cirrhotic rats was significantly increased, however, the Hyp content was marked reduced in ALR-treated rats, compared with cirrhotic rats. In conclusion, these results provided novel insights into the mechanisms of ALR in the protection of the liver.
研究发现在各种急、慢性肝损伤动物模型中,ALR可促进肝细胞再生,减轻肝损伤,改善肝功能,增加动物存活率和存活时间。本文采用尾静脉注射pcDNA3.1-hALR重组质粒治疗毒物诱导和免疫性大鼠肝纤维化。
研究结果显示,ALR基因治疗可明显减轻大鼠肝损伤和肝纤维化,其机制可能是抑制肝细胞内氧化应激,下调各种致纤维化因子,阻止肝星状细胞的活化。ALR治疗可显著减少α-SMA的表达,减少肝内活性氧簇水平,并下调TGF-β1、PDGF-BB和TIMP-1的表达。与肝硬化模型组大鼠相比,治疗组大鼠的血清AST, ALT和AKP水平明显降低,但是,血清ALB明显高于模型组大鼠而接近健康对照组。血清生化指标的变化与肝组织病理检查结果相符,治疗组大鼠肝组织H&E染色显示肝小叶结构的破坏得到修复,Sirius-red胶原特异性染色显示胶原阳性面积显著减少,qRT-PCR结果表明Ⅰ型胶原和Ⅲ型胶原含量明显减少。与健康对照组比较,模型组大鼠肝脂质过氧化指标MDA含量增加,SOD活性降低。经过ALR治疗后,治疗组大鼠MDA含量降低,SOD活性增加。与健康对照组相比,模型组大鼠肝脏Hyp含量显著增加。而与模型组大鼠比较,治疗组大鼠肝脏Hyp含量明显减少。总之,研究结果提供了新的ALR保护肝脏的机制。
Liver fibrosis is a common pathological process of chronic hepatic disease, which can lead to cirrhosis and increase risk for hepatocellular carcinoma. In China, liver fibrosis and cirrhosis are usually caused by the developing of chronic viral hepatitis. Here we evaluated the therapeutic efficacy of augmenter of liver regeneration (ALR) on liver fibrosis induced by carbon tetrachloride (CC14) and porcine-serum (PS), and investigated potential mechanisms.
Accumulated studies had revealed that ALR appeared to be an important regulator of liver regeneration and had trophic effects on damaged liver in animal models of acute or chronic liver injury. In these studies, exogenous ALR administration ameliorated hepatic damage, enhanced hepatic proliferative capacity post-toxininduced liver injury, or improved survival of toxin-administered animals. The present research was to observe the therapeutic effects of ALR on liver fibrosis and to explore the molecular mechanism of antifibrosis.
ALR might attenuate liver injury and fibrosis by suppressing oxidative stress, down-regulating profibrogenic factors, and blocking HSCs activation. This report demonstrated that ALR therapy diminished α-smooth muscle actin (SMA) expression, decreased intrahepatic reactive oxygen species (ROS) level, and down-regulated transforming growth factor (TGF)-β1, platelet-derived growth factor (PDGF)-BB, and tissue inhibitor of metalloproteinase (TIMP)-1expression. Compared with the model rats, cirrhotic rats treated with ALR showed ameliorated biochemical liver tests. In these animals, serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (AKP) were significantly lower and serum albumin (ALB) significantly higher than in cirrhotic rats and similar to healthy controls. These favorable changes were accompanied by histological improvement with marked reduction of fibrosis and decreased expression of collagen I and IV in ALR-treated rats. Compared with cirrhotic rats, ALR treatment were able to reduce the level of malondialdehyde (MDA) content, increased superoxide dismutase (SOD) activity. Compared healthy control rats, the Hyp content of cirrhotic rats was significantly increased, however, the Hyp content was marked reduced in ALR-treated rats, compared with cirrhotic rats. In conclusion, these results provided novel insights into the mechanisms of ALR in the protection of the liver.
引文
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