地参酚类化合物对肝损伤的保护作用及机制研究

In consequence of environmental pollution, poor dietary habits, occupational exposure, disease and autoimmunity, the population with liver damage are increasing, which adds huge burden to the individuals, families and states. Medical treatments for liver diseases are often difficult to manage and have limited efficacy. It has been an important way to obtain efficient and natural hepatoprotective compounds from plant materials. Liver damage usually accompanies with oxidative stress. Phenolic compounds are potential natural and efficient hepatoprotective medicines duo to their strong antioxidant activity. Lycopus lucidus Turcz. is a perennial member of the Labiata family with highly nutritional and healthy effects. In this project, the in vivo and in vitro carbon tetrachloride-induced liver damage model will be built, and the hepatoprotective effect of phenolic compounds from Lycopus lucidus Turcz. will be established on the basis of relevant indicators of liver function. Additionally, the target of phenolic compounds will be searched from the aspect of oxidative stress, inflammatory reaction and the change of mitochondria, and the hepatoprotective mechanism will be revealed from celluar and molecular level. The results of the research have a very important practical significance for the development of natural hepatoprotective medicines, utilization and development of Lycopus lucidus Turcz. and promotion of the local economics.

由于环境的污染、不良的饮食习惯、职业接触、疾病、自身免疫等原因，肝损伤人群日益增多，给个人、家庭和社会带来极大的经济负担。药物治疗肝损伤管理起来通常有困难，而且效果也有限，从植物原料中获取天然高效的保肝物质成为了一个重要的途径。酚类化合物具有强的抗氧化活性，而肝损伤通常伴随着氧化应激，因此酚类化合物是潜在的天然高效保肝药物。地参（Lycopus lucidus Turcz.），系唇形科(Labiatae) 泽兰属多年生草本植物，具有很高的营养及保健功效。本项目拟通过建立体内、体外四氯化碳肝损伤模型，依据肝功能相关指标，明确地参酚类化合物对四氯化碳肝损伤的保护作用，并从氧化应激、炎症反应和线粒体变化的角度，寻找地参酚类化合物的作用靶点，从细胞及分子水平揭示其保肝机制。研究结果对于天然保肝药物的开发，地参资源的开发与利用以及促进当地经济的发展具有重要的现实意义。

地参（Lycopus lucidus Turcz.），系唇形科(Labiatae) 泽兰属多年生草本植物，具有很高的营养及保健功效，通过建立体内、体外CCl4肝损伤模型，研究地参酚类化合物对CCl4肝损伤的保护作用，并从氧化应激、炎症反应和线粒体变化的角度揭示其保肝机制。 体外研究结果显示，地参游离酚和结合酚提取物在0.4-1.6 mg/mL范围内，可显著抑制CCl4所致BRL大鼠肝细胞损伤AST、ALT、LDH、ROS、8-OHdG、Cyt-C和Caspase-3含量的升高，升高SOD、GSH的水平，降低MDA、TNF-α、IL-6、IL-8、iNOS和NO的含量，并抑制细胞凋亡及影响细胞周期；地参主要酚类化合物单体迷迭香酸和咖啡酸在0.2-0.8 mg/mL范围内，可显著抑制CCl4所致BRL大鼠肝细胞损伤AST、ALT、LDH、ROS、Cyt-C和8-OHdG水平的升高，上调GSR和SOD mRNA的表达，降低COX-2、iNOS、IL-6和Caspase-3的表达水平。体内研究结果显示，地参游离酚（100 mg/kg bw）具有显著的CCl4肝损伤保护作用，能显著降低CCl4所致血清ALT、AST、ALP、TG、TC、TBIL的升高，升高肝脏GSH、CAT、SOD水平，降低MDA含量；病理学检验结果显示，地参游离酚组小鼠肝细胞排列相对整齐，胞质均匀清晰，只有部分细胞点状坏死，无大面积的凝固性坏死；Tunnel染色结果表明，地参游离酚能抑制CCl4所致小鼠DNA的断裂；免疫组化实验结果显示，地参游离酚能显著降低TNF-α、IL-6、IL-8、iNOS、COX-2、Caspase-3的水平；迷迭香酸高剂量组（40 mg/kg bw）具有显著的CCl4肝损伤保护作用，与模型组相比，肝脏和脾脏指数显著降低，血清ALT、AST、ALP、TG、TC、TBIL显著降低，肝脏GSH、CAT、SOD水平显著升高，MDA、NO、ROS、8-OHdG、Cyt-C、IL-6、IL-8、TNF-α含量显著降低；病理学检验结果进一步证实了迷迭香酸对CCl4肝损伤的保护作用。此外，研究了两个采收地三个采收期地参游离酚和结合酚提取物体外对消化酶的抑制活性。