Objective: To study and explore the effect and mechanism of action of Jieduhuayu granules on oxidative injury of human liver L02 cells. Methods: Human liver L02 oxidative injury model was established with 0.1 mmol/ L H(2)O(2) intervention for 1 h, and treated with different concentrations of Jieduhuayu (JDHY) solution. Hepatocytes were divided into five groups: normal, H(2)O(2), H(2)O(2) + JDHY (0.5 mg/ml), H(2)O(2) + JDHY (1 mg/ml), and H(2)O(2) + JDHY (1.5 mg/ml). MTT assay was used to detect hepatocytes activity. Transmission electron microscope was used to observe mitochondrial morphology in hepatocytes. Biochemical test was used to detect the levels of superoxide dismutase, lactate dehydrogenase, alanine aminotransferase, aspartate aminotransferase, malondialdehyde, and reduced glutathione and albumin in hepatocytes. Western blot was used to detect the expression levels of rabbit anti-phosphatidylinositol 3-kinase (PI3K), AKT and mTOR in hepatocytes. One-way analysis of variance was used for comparison between multiple groups, and the LSD method was used for pairwise comparison. Results: Compared with the normal group, the cell proliferation activity (P < 0.05), mitochondrial vacuolization, superoxide dismutase activity, reduced albumin and glutathione content, and PI3K, AKT, and mTOR protein expression levels in the H(2)O(2) group were all significantly reduced (P < 0.05), while the content of malondialdehyde and the activities of alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase were significantly increased (P < 0.05). Compared with H(2)O(2) group, the cell proliferation activity (P < 0.05), alterations in morphological remission of mitochondria, superoxide dismutase activity, reduced albumin and glutathione content, and PI3K, AKT and mTOR protein expression levels in the H(2)O(2) + JDHY (1 mg/ml) and H(2)O(2) + JDHY (1.5 mg/ml) group (P < 0.05) were all significantly increased (P < 0.05), while malondialdehyde content and alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase activities were significantly decreased (P < 0.05). Conclusion: Jieduhuayu granule can effectively improve oxidative stress and mitochondrial injury in hepatocytes, and its effect may be related to the promoting expression of PI3K/AKT/mTOR signaling pathways.
目的： 研究解毒化瘀颗粒对氧化损伤人肝细胞L02的影响，并探讨其作用机制。 方法： 采用0.1 mmol/L H(2)O(2)干预1 h构建人肝细胞L02氧化损伤模型，并采用不同浓度的解毒化瘀工作液（Jie-Du-Hua-Yu, JDHY）进行处理。将细胞分为5组：正常组、H(2)O(2)组、H(2)O(2) + JDHY（0.5 mg/ml)组、H(2)O(2) + JDHY (1 mg/ml)组、H(2)O(2) + JDHY (1.5 mg/ml)组。MTT检测细胞活性，透射电镜观察细胞线粒体形态，生物化学检测细胞内超氧化物歧化酶、乳酸脱氢酶、丙氨酸转氨酶、天冬氨酸转氨酶、丙二醛、还原型谷胱甘肽和白蛋白水平，蛋白质印迹检测细胞内兔抗磷脂酰肌醇3-激酶（PI3K）、AKT、mTOR表达水平。多组间比较采用单因素方差分析，两两比较采用LSD法。 结果： 与正常组比较，H(2)O(2)组细胞增殖活性显著降低（P < 0.05），细胞线粒体空泡化，超氧化物歧化酶活性、白蛋白、还原型谷胱甘肽含量以及PI3K、AKT、mTOR蛋白表达水平均显著降低（P值均< 0.05），而丙二醛含量和丙氨酸转氨酶、天冬氨酸转氨酶及乳酸脱氢酶活性显著升高（P值均< 0.05）。与H(2)O(2)组比较，H(2)O(2) + JDHY (1 mg/ml)组和H(2)O(2) + JDHY (1.5 mg/ml)组细胞增殖活性显著升高（P值均< 0.05），线粒体形态变化得到缓解，超氧化物歧化酶活性、白蛋白、还原型谷胱甘肽含量以及PI3K、AKT、mTOR蛋白表达水平均显著升高（P值均< 0.05），丙二醛含量和丙氨酸转氨酶、天冬氨酸转氨酶及乳酸脱氢酶活性显著降低（P值均< 0.05）。 结论： 解毒化瘀颗粒可有效改善肝细胞氧化应激与线粒体受损，其作用可能与促进PI3K/AKT/mTOR信号通路表达有关。.
Keywords: Jie-Du-Hua-Yu granules; Mitochondrion; Oxidative stress; PI3K/AKT/mTOR signaling pathway.