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目的:观察溪黄草总二萜的保肝作用。方法:制备乙醇损伤L-02细胞模型,考察总二萜对受损细胞ALT、AST泄漏量的影响。复制两种急性肝损伤动物模型,分离血清检测转氨酶活性,取肝组织进行病理学观察。提取肝脏线粒体检测MMP、MPTP开放程度、Mn-SOD活力、8-OHd G水平、呼吸链复合物Ⅰ、复合物Ⅲ活性。肝匀浆测定MDA水平和GSH、GSH-Px活力。RT-PCR检测大鼠肝组织中Nrf2、Bach1、HO-1 mRNA表达水平;Western Blot检测大鼠肝组织HO-1总蛋白的表达水平。结果:细胞实验显示,溪黄草总二萜5.00、10.00、20.00μg/ml给药组有降低受损L-02细胞转氨酶泄漏的趋势。动物实验显示,溪黄草总二萜25.0、50.0、100.0 mg/kg治疗组动物血清转氨酶水平均明显下降,肝细胞水肿减轻,病理分级下降。溪黄草总二萜25.0、100.0 mg/kg治疗组动物肝脏线粒体Mn-SOD、ComplexⅠ、ComplexⅢ活性上升;50.0 mg/kg治疗组MPTP开放被抑制、8-OHd G水平下降。溪黄草总二萜25.0、50.0、100.0 mg/kg治疗组动物肝匀浆中MDA水平下降,GSH、GSH-Px活力升高。溪黄草总二萜50.0、100.0 mg/kg治疗组大鼠肝组织中HO-1 mRNA、总蛋白表达水平上升。结论:溪黄草总二萜在细体内外实验中显示均具有明确的保肝作用,其保肝作用与保护肝脏线粒体、调控肝脏内源性抗氧化酶系统有关。
Objective: To observe the hepatoprotective effect of total diterpene of Xihuangcao. Methods: L-02 cell model of ethanol injury was prepared to investigate the effect of total diterpene on the amount of ALT and AST in damaged cells. Two animal models of acute liver injury were duplicated. Serum transaminase activity was detected by sera. Liver tissues were harvested for pathological observation. The liver mitochondria were extracted for detection of MMP, MPTP opening degree, Mn-SOD activity, 8-OHd G level, respiratory chain complex Ⅰ, complex Ⅲ activity. Liver homogenate MDA levels and GSH, GSH-Px activity. The expression of Nrf2, Bach1, HO-1 mRNA in rat liver tissue was detected by RT-PCR. The expression of HO-1 protein in liver tissue was detected by Western Blot. Results: The cell experiments showed that the total diterpene 5.00,10.00,20.00μg / ml administration group of Xihuangcao could reduce the leakage of transaminase in the damaged L-02 cells. Animal experiments showed that total serum diterpenoid 25.0,50.0,100.0 mg / kg Xihuangcao serum transaminase levels were significantly decreased, hepatocyte edema reduced, the pathological grade decreased. The activities of Mn-SOD, ComplexⅠ and ComplexⅢ in hepatic mitochondria increased in 25.0,100.0 mg / kg xanthophyll group total diterpene treatment group. The MPTP opening was inhibited and the level of 8-OHd G decreased in 50.0 mg / kg treatment group. The total diterpene 25.0,50.0,100.0 mg / kg in the treatment group decreased MDA level in the liver homogenate, GSH, GSH-Px activity increased. The content of HO-1 mRNA and total protein in the liver tissue of 50 mg / kg and 100.0 mg / kg total diterpene in Xihuangcao increased. CONCLUSION: The total diterpene in Xihuangcao has a clear hepatoprotective effect both in vitro and in vivo. The hepatoprotective effects of the two diterpenes are related to protecting the mitochondria of the liver and regulating the endogenous antioxidant enzyme system in the liver.