【作者】 齐艳萍；

【导师】 李和平；

【作者基本信息】 东北林业大学 ， 野生动植物保护与利用， 2008， 博士

【摘要】 急性肝损伤是临床常见、多发的肝脏病患,是各种肝病共同的病理基础,其病理特点主要为肝细胞变性、坏死,甚至凋亡,其长期存在往往导致肝纤维化,甚至是肝硬化、肝癌发生的重要的使动因素。因此防治肝损伤是临床肝病治疗的主要环节之一,及时控制肝损伤的发生和发展对治疗肝病将有重要的临床意义。鹿茸含有多种生物活性成分,有广泛的药理作用和医疗保健功能。在现代医疗实践中,以鹿茸为主要原材料制作的药剂已经应用于多种疾病的治疗,并取得良好的效果,但在肝病治疗中尚未得以普遍应用。如果将鹿茸应用到肝病的治疗和保健中,不仅可解决医学上一大难题,而且也可以为鹿茸的开发带来巨大的经济效益和社会效益。目前研究已经揭示鹿茸对肝损伤确有一定的修复作用,但对其抗肝损伤的具体作用及作用机理尚不十分清楚。本研究以小鼠为实验对象,利用CCl₄、APAP、56°白酒以及BCG+LPS几种毒物制造化学性、药物性、酒精性和免疫性四种不同类型的急性肝损伤模型,通过监测血清中ALT、AST、TP、ALB、TBil、TG的浓度和肝组织匀浆中MDA、SOD、NO、ADH的含量,来研究以马鹿茸为原料制备的鹿茸粉和鹿茸醇提物对不同类型肝损伤的修复作用及相关机理,旨在从肝细胞损伤、肝脏合成贮备功能、分泌排泄功能、脂代谢功能及脂质过氧化几个方面阐明鹿茸对肝脏的保护作用及部分作用机理,为充分开发鹿茸新的药用价值,研制鹿茸抗肝损伤的新型医疗保健药物提供生化药理学基础。1.鹿茸对CCl₄所致化学性肝损伤修复作用的研究结果表明:对肝细胞损伤的保护作用方面,鹿茸醇提物1.0g/kg、2.0g/kg均可以明显降低ALT、AST的含量（P＜0.01）,其中醇提物2.0g/kg将ALT由204.08±72.00U/L降至55.21±7.93U/L,AST由234.77±7.71U/L降至80.57±10.80U/L,其降酶作用与联苯双酯（71.25±14.06、79.83±11.02U/L）相当。在提高肝脏蛋白合成储备功能方面,鹿茸醇提物1.0g/kg、2.0g/kg均能明显提高血清TP（56.94±5.96、59.90±3.89g/L）及ALB（31.80±0.92、31.30±0.72g/L）含量（P＜0.01）,作用效果优于联苯双酯（37.94±5.80、29.18±2.78g/L）,而鹿茸粉效果不显著。对肝脏排泄分泌功能的改善方面,鹿茸醇提物2.0g/kg能将血清TBil明显降低至6.35±0.22μmol/L（P＜0.01）,效果与联苯双酯（6.33±0.29μmol/L）相当,而鹿茸醇提物1.0g/kg稍差（P＜0.05）。改善肝脏脂代谢功能方面,各治疗组均能降低血清TG含量,其中鹿茸醇提物2.0g/kg可将TG由1.03±0.09mmol/L降至0.70±0.12mmol/L,作用效果明显（P＜0.05）且与联苯双酯（0.87±0.06mmol/L）相当。在对抗肝脏自由基的脂质过氧化方面,鹿茸醇提物能显著提高SOD活力、降低MDA和NO水平（P＜0.01）,鹿茸醇提物2.0g/kg作用效果稍好,可将SOD活力提高至78.75±0.56U/mgprot,MDA和NO含量降低至0.93±0.07nmol/mg和40.82±3.73umol/L,而鹿茸粉效果较差。此外,鹿茸能显著降低肝损伤小鼠的肝重系数（P＜0.01）,醇提物1.0g/kg组肝重系数为5.01±0.12,其作用效果较2.0g/kg组（5.16±0.07）稍好,但差异不明显。2.鹿茸对APAP所致药物性肝损伤修复作用的研究结果表明:对受损肝细胞的保护作用方面,鹿茸醇提物2.0g/kg将ALT、AST分别降至38.38±2.19U/L、36.75±0.92U/L,作用效果优于1.0g/kg（41.67±11.56U/L、56.67±8.09U/L）。在蛋白合成储备功能方面,各治疗组TP含量均有小幅升高,其中醇提物1.0g/kg组TP升高至38.17±0.76g/L,鹿茸粉组升高至27.17±0.61g/L,两组效果均显著（P＜0.05）,而ALB含量升高幅度较大,醇提物2.0g/kg治疗效果最好（P＜0.01）,其ALB含量高达29.86±1.00g/L。在改善肝脏的胆汁瘀滞程度方面,各治疗组TBil含量均明显降低（P＜0.01）,其中鹿茸醇提物2.0g/kg可将TBil降低至8.59±0.27μmol/L,作用效果优于醇提物1.0g/kg,但均低于联苯双酯（8.27±0.38μmol/L）。对脂代谢功能的调节方面,各治疗组TG含量都有大幅度的降低（P＜0.01）,其中鹿茸醇提物2.0g/kg的疗效最好,其TG由1.68±0.05mmol/L降至0.86±0.03mmol/L,低于联苯双酯（0.61±0.02mmol/L）。鹿茸各治疗组均能在一定程度上提高SOD活力,降低NO和MDA水平,抑制APAP所致受损肝脏的脂质过氧化反应,但均未达到联苯双酯组中SOD（77.56±2.18U/mgprot）、MDA（0.77±0.06nmol/mg）和NO（45.24±4.39umol/L）水平。鹿茸还能能显著降低APAP急性肝损伤小鼠的肝重系数,其中醇提物2.0g/kg组与模型组相比差异极显著（P＜0.01）,将肝重系数由6.52±0.23降至5.64±0.11,作用效果较1.0g/kg的明显。3.鹿茸对酒精性肝损伤修复作用的研究结果表明:对减轻肝细胞的损伤程度方面,各组ALT、AST均有不同程度的降低,其中鹿茸醇提物2.0g/kg对AST的降低效果极显著（P＜0.01）,可将AST由56.00±2.31U/L降至36.42±1.45U/L,结合ALT和AST综合判断,鹿茸醇提物2.0g/kg效果最佳。对改善肝脏的合成储备功能方面研究发现,鹿茸对促进TP的合成作用不显著,而能显著增加肝脏对ALB的合成功能（P＜0.01）,鹿茸醇提物2.0g/kg作用效果较好,能将其提高至28.71±1.12g/L,仅次于联苯双酯的29.15±0.95g/L。在肝脏对TBil的摄取、结合和排泄能力方面,鹿茸缓解急性酒精肝损伤小鼠肝脏胆汁瘀滞的程度,给药后各组TBil含量均大幅降低（P＜0.01）,其中鹿茸醇提物1.0g/kg作用效果最好,使TBil达到10.99±1.55μmol/L,仅次于联苯双酯（9.46±0.11μmol/L）。鹿茸能有效抑制脂肪变性,调整脂代谢功能,鹿茸醇提物1.0g/kg组和鹿茸粉组可将TG降低至0.80±0.07mmo/L和0.94±0.02mmol/L,作用效果较差,而鹿茸醇提物2.0g/kg能将TG降低至0.65±0.28mmol/L,作用效果极显著（P＜0.01）,且与联苯双酯（0.61±0.07mmol/L）相当。在抗急性酒精肝损伤脂质过氧化方面,鹿茸粉作用效果低于鹿茸醇提物,其中醇提物2.0g/kg效果最好,可将SOD活性提高至73.20±0.60U/mgprot,MDA和NO含量降低至1.05±0.07nmol/mg和50.00±3.19umol/L,但均低于联苯双酯。在肝脏对酒精的代谢能力方面,鹿茸能明显增加肝脏ADH活性,与模型组相比,鹿茸粉可使ADH活性由0.93±0.03U/mg提高至1.11±0.04U/mg（P＜0.05）,其作用效果明显低于鹿茸醇提物1.0g/kg组的1.42±0.06U/mg和2.0g/kg组的1.52±0.03U/mg（P＜0.01）。此外,鹿茸能有效抑制大量饮酒造成的肝脏重量的增加,鹿茸醇提物2.0g/kg作用效果最好（P＜0.01）,将肝重系数降至4.99±0.23,作用与联苯双酯相当。4.鹿茸对BCG和LPS联合所致免疫性肝损伤修复作用的研究结果表明:对肝细胞损伤的保护作用方面,鹿茸给药后ALT、AST均不同程度降低,鹿茸醇提物2.0g/kg可将ALT、AST分别由43.79±1.51U/L、53.50±1.72U/L降至33.33±2.78U/L、39.50±0.93U/L,其对于免疫性肝损伤的保护作用较1.0g/kg稍好而与联苯双酯相当,鹿茸粉（37.17±2.20U/L、50.83±5.92U/L）作用最差。在促进肝脏蛋白质的合成代谢功能方面,各组TP和ALB均有不同程度的提高,鹿茸醇提物效果高于鹿茸粉,其中醇提物2.0g/kg效果较好,可将TP、ALB分别提高至36.81±0.55g/L、26.62±0.54g/L,但低于联苯双酯（38.35±1.31g/L、27.61±0.65 g/L）。在改善摄取、转化和排泄TBil的能力方面,给药后各组TBil含量均极显著降低（P＜0.01）,鹿茸醇提物2.0g/kg将TBil由17.42±0.74μmol/L降低至8.82±0.20μmol/L,作用效果优于其它组,仅次于联苯双酯（7.85±0.51μmol/L）。在调节脂代谢功能方面,各组TG含量在用药后均明显降低,鹿茸醇提物2.0g/kg降低幅度最大（P＜0.01）,由1.23±0.07mmo/L降至0.87±0.03mmo/L,效果较联苯双酯（0.79±0.03mmo/L）稍差。对抑制脂质过氧化方面,鹿茸粉和鹿茸醇提物1.0g/kg提高SOD活力的效果较好,可使SOD值达到75.67±1.63U/mgprot和75.57±1.88U/mgprot,鹿茸醇提物降低MDA的效果优于鹿茸粉,其中醇提物2.0g/kg（1.22±0.07nmol/mg）效果略高于1.0g/kg（1.39±0.10 nmol/mg）,而NO的降低效果以鹿茸醇提物1.0g/kg效果最为理想,由61.22±5.14umol/L降低至40.82±6.56umol/L,根据MDA、SOD和NO综合分析,鹿茸醇提物1.0g/kg抗脂质过氧化反应的效果较好,但较联苯双酯稍差。此外,鹿茸能抑制免疫性肝损伤造成的肝脏重量的增加,鹿茸粉效果最差,仅由6.22±0.10降至5.97±0.07,鹿茸醇提物2.0g/kg效果较好（P＜0.01）,可将达5.74±0.10,但效果低于联苯双酯（5.61±0.11）。更多还原

【Abstract】 The acute liver damage was a kind of common pathological changes of various liver diseases.A characteristic feature of the pathology was hepatocyte degeneration,necrosis, even apoptosis.Its long-term existence always leaded to hepatic fibrosis,cirrhosis and hepatoma.Therefore,preventing and curing liver damage were the key points of treating clinical liver diseases.Controlling the liver damage in time was particularly important to cure the liver diseases.Velvet has extensive pharmacological action and medical care function.In modern medical practice,the medicines with velvet as its main raw materials gained good effects on treating many diseases.But velvet wasn’t used to treat the liver disease.If velvet could be applied to the treatment of liver diseases and the health care,that it could not only resolve a difficult medical problem,but also bring huge economic benefit and the social effect.Some studies showed that velvet had certain repair action for the liver damage,but the action mechanism was still not very clear.In this study,we dealt the rats with CCl₄、APAP、alcohol and BCG+LPS,established the model of acute liver damage,such as chemical,medicamentosa and immune liver damage.By measuring the content of ALT、AST、TP、ALB、TBil、TG in serum and MDA、SOD、NO、ADH in liver homogenate,expounded the effects and mechanism of velvet powder and the EAV（extract by alcohol from velvet） for the liver damage,finally it could build the foundation of biochemistry and pharmacology for anti-liver damage of velvet.1.The main results obtained from studies on the repair effects of velvet for the chemical liver damage are as follows:In terms of protecting the liver cell damage,the EAV（1.0g/kg and 2.0g/kg） could reduce the content of ALT and AST in serum（P＜0.01） obviously.The rats were treated with EAV（2.0g/kg）,then the content of ALT was reduced from 204.08±72.00U/L to 55.21±7.93U/L,and the content of AST was reduced from 234.77±7.71U/L to 80.57±10.80U/L;the same effects existed between BIF（71.25±14.06、79.83±11.02U/L ） and EAV（2.0g/kg）.In terms of enhancing the function of protein synthesis and reserve, EAV（1.0g/kg and 2.0g/kg） could increase the content of TP（56.94±5.96、59.90±3.89g/L） and ALB（31.80±0.92、31.30±0.72g/L） in serum obviously（P＜0.01）,the protective effects was superior to the BIF（37.94±5.80、29.18±2.78g/L）;but the effects of velvet powder was not prominent.In terms of improving the liver excretory ability,EAV（2.0g/kg） could reduce the content of TBil（6.35±0.22μmol/L） obviously（P＜0.01） obviously,its effects were as good as BIF（6.33±0.29μmol/L）;but the EAV（1.0g/kg） was a little worse（P＜0.05）.In terms of improving the liver fat metabolism,the content of TG in each treatment group were reduced;the EAV（2.0g/kg） could reduce the content of TG form 1.03±0.09mmol/L to 0.70±0.12mmol/L,the same effects occurred between BIF（0.87±0.06mmol/L） and EAV（2.0g/kg）.For reducing lipid peroxidation of free radicals in liver,the EAV could enhance the vitality of SOD obviously,and reduce the level of MDA and NO（P＜0.01）;the EAV（2.0g/kg） could increase the content of SOD（78.75±0.56U/mgprot）,and reduce the content of MDA（0.93±0.07nmol/mg） and NO（40.82±3.73umol/L）;but the effects of velvet powder was bad.Besides these,velvet could reduce the rat’s liver weight coefficient obviously（P＜0.01）,but the difference was not significant in each group.2.The main results obtained from studies on the repair effects of velvet for the medicamentosa liver damage by APAP are as follows:For protecting the liver’s damaged cell,EAV（2.0g/kg） could reduce the content of ALT（38.38±2.19U/L） and AST（36.75±0.92U/L）,its effects were better than that of EAV（1.0g/kg）.For enhancing the function of protein synthesis and reserve,the content of TP in each treatment group had a small rise;the difference between the effects（38.17±0.76g/L） of EAV（1.0g/kg）and that（27.17±0.61g/L） of velvet powder was significant;the content of ALB had a big rise,EAV（2.0g/kg） remarkable enhanced ALB’s content（29.86±1.00g/L）,its effects were the best（P＜0.01）.For improving cholestasis of liver,the content of TBil reduced obviously,the effects of BIF（8.27±0.38μmol/L） were better than that（8.59±0.27μmol/L） of EAV（2.0g/kg）.For adjusting lipid metabolism,the level of TG in each treatment group was reduced a lot（P＜0.01）.EAV（2.0g/kg） reduced TG’s level from 1.68±0.05mmol/L to 0.86±0.03mmol/L,the level was lower than BIF（0.61±0.02mmol/L）.Velvet could enhance the vitality of SOD in each treatment group, and reduce the level of NO and MDA;inhibit the lipid per-oxidization of damaged liver of rats dealt with APAP,but their effects were worse than BIF.Velvet could reduce the rat’s liver weight coefficient obviously,EAV（2.0g/kg） was the better than EAV（1.0g/kg）,it could reduce the liver weight coefficient from 6.52±0.23 to 5.64±0.11.3.The main results obtained from studies on the repair effects of velvet for the alcoholic liver damage are as follows:For alleviating the liver’s damaged cell,the levels of ALT、AST all reduced, EAV（2.0g/kg） reduced the content of AST from 56.00±2.31U/L to 36.42±1.45U/L very significantly,its effects were the best.For enhancing protein synthesis and reserve of liver, velvet could not improved TP synthesis remarkably,but could improved ALB synthesis evidently（P＜0.01）;but the remedial ability of EAV（2.0g/kg） was worse than BIF（29.15±0.95g/L）.For adjusting the ability of liver with TBil ingested、combined and excreted,velvet could alleviate cholestasis of liver,the content of TBil was reduced in each treatment group obviously（P＜0.01） after dealt with velvet;the effects of BIF（9.46±0.11μmol/L） were better than EAV（1.0g/kg）,was the best.Velvet could inhibit fatty degeneration and adjust its metabolism;the effects were good by BIF（0.61±0.07mmol/L） and EAV（2.0g/kg）,the effects surpassed EAV（1.0g/kg） and velvet powder（0.94±0.02mmol/L） in remedial ability.For acuted lipid per-oxidation by alcohol in damaged liver,remedial ability of EAV（1.0g/kg、2.0g/kg） was better than velvet powder,EAV（2.0g/kg） could adjust the vitality of SOD（73.20±0.60U/mgprot）,and reduce the level of MDA（1.05±0.07nmol/mg） and NO（50.00±3.19umol/L）;but BIF had the best effects.For alcoholic metabolism in liver,the vitality of ADH was enhanced by velvet; EAV（1.0g,/kg、2.0g/kg） could enhanced the vitality of ADH（1.42±0.06U/mg、1.52±0.03U/mg）,their effects both surpassed velvet powder（1.11±0.04U/mg）.Velvet could effectively inhibit liver weight increasing caused by alcohol;EAV（2.0g/kg） reduced the liver weight coefficient to 4.99±0.23（P＜0.01）,have as good effects as BIF.4.The main results obtained from studies on the repair effects of velvet for the immune liver damage by APAP are as follows:For protecting the liver’s damaged cell,velvet could reduce content of ALT、AST; effects of BIF and EAV（2.0g/kg） were better,that of velvet powder（ALT:37.17±2.20U/L、AST:50.83±5.92U/L）was the worst.For enhancing the function of protein synthesis and reserve,the levels of TP、ALB were increased;the effects of BIF（38.35±1.31 g/L、27.61±0.65 g/L） was the best,and surpassed EAV;velvet powder was the worst.For adjusting the ability of liver with TBil ingested、combined and excreted,the level of TBil was increased in all groups（P＜0.01）;the effects of BIF（7.85±0.51μmol/L） were the best,and surpassed EAV（2.0g/kg）.For fatty metabolism in liver,TG’s level was reduced obviously with velvet;EAV（2.0g/kg） could reduce the content of TG from 1.23±0.07mmo/L to 0.87±0.03mmo/L（P＜0.01）,the effects were worse than BIF.For inhibiting lipid per-oxidation,the effects of heightening the vitality of SOD by velvet powder and EAV（1.0g/kg） were better,it was enhanced to 75.67±1.63U/mgprot and 75.57±1.88U/mgprot;the remedial effects of BIF were the best,and it was better than EAV（1.0g/kg） for reducing the level of MDA、NO.Velvet could obviously inhibit liver weight increasing（P＜0.01） caused by the immune liver damage;the effects of BIF（5.61±0.11）were the best,and EAV（2.0g/kg） was worse,the effects of velvet powder were the worst.更多还原