五酯滴丸保肝作用机制研究

Study of the action mechanism of Wuzhi Diwan's protective effect on liver

目的以金属硫蛋白-Ⅰ/Ⅱ基因敲除的MT^((-/-))小鼠和同源野生型MT^((+/+))小鼠为研究对象,建立APAP肝损伤模型,从氧化损伤方面研究五酯滴丸的保肝作用及其机制。方法 MT^((+/+))小鼠和MT^((-/-))小鼠分别给予五酯滴丸(20mg/kg)灌胃,每天给药1次,连续给药7天。末次给药后24 h给予500 mg/kg APAP处理,观察血清中丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转酶(AST)、乳酸脱氢酶(LDH)、嘌呤核苷磷酸化酶(PNP)和苹果酸脱氢酶(MDH)的表达水平;肝组织中丙二醛(MDA)、谷胱甘肽过氧化物酶(GPx)、超氧化物歧化酶(SOD)含量以及肝组织病理学;Western Blot法观察五酯滴丸对MT的调节作用。结果五酯滴丸预处理可有效改善APAP造成的脂肪变性和坏死等病理学改变,有效降低血清ALT、AST、LDH、PNP和MDH表达,与APAP模型组相比,差异显著。五酯滴丸能够降低MDA含量,增高GPx和SOD水平,具有一定的抗氧化损伤作用,且对MT^((+/+))小鼠作用更明显。五酯滴丸预处理能够诱导MT^((+/+))小鼠中MT的表达,减轻肝损伤。五酯滴丸预处理对MT^((-/-))小鼠肝损伤无明显保护作用。结论五酯滴丸预处理能够提高MT表达,对APAP诱发的MT^((+/+))小鼠肝损伤具有明显的保护作用。提示MT的高表达是五酯滴丸肝脏保护作用的可能机制之一。

Objective To take metallothione-Ⅰ/Ⅱ knockout MT（＋/＋）mice and MT（-/-）mice as the research object to build the model with liver injury in order to research on Wuzhi Diwan’s protective effect on liver mainly from the aspects of oxidative damage. Methods MT（＋/＋）mice and MT（-/-）mice were respectively treated with Wuzhi Diwan（ 20 mg / kg） through intragastric administration once a day for 7 days successively. And then 24 h after the last administration,they were processed with APAP（ 500 mg / kg）. Some indicators,such as alanine aminotransferase（ ALT）, aspartate aminotransferase（ AST）, lactate Dehydrogenase（ LDH）, purine nucleoside phosphorylase（ PNP）,malate dehydrogenase（ MDH）,lipid peroxidation（ MDA）,glutathione peroxidase（ GPx）,superoxide（ SOD）and liver tissue in pathology were observed. Western Blot was used to observe the adjustment role of Wuzhi Diwan on MT. Results The pretreatment with Wuzhi Diwan could effectively improve the adipose degeneration and necrosis caused by APAP. The expression levels of ALT,AST,LDH, PNP and MDH were effectively decreased and the differences were obvious compared with APAP model group. Wuzhi Diwan could decrease the level of MDA while increase the levels of GPx and SOD so as to have an action of anti-oxidation injury with better effect on MT（-/-）mice. The pretreatment with Wuzhi Diwan could induce the expression of MT in MT（＋/＋）mice to alleviate liver injury. The pretreatment with Wuzhi Diwan had no protective function on liver injury in MT（-/-）mice. Conclusion The pretreatment with Wuzhi Diwan can enhance the expression of MT and has a protective effect on liver injury induced by APAP. This gives a hint that high-expression of MT maybe one of the mechanism of protective action of Wuzhi Diwan on liver.