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目的:建立使用微载体进行LO2肝细胞系高密度培养的方法,探讨雷公藤内酯醇的肝毒性作用及机制。方法:在限制贴壁的条件下,以Cytodex-3为材料进行LO2肝细胞的三维立体培养,在光镜下观察其生长情况,探究雷公藤内酯醇对肝细胞增殖能力的影响,并对细胞上清液的丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)和丙二醛(MDA)以及裂解细胞样品的谷胱甘肽过氧化物酶(GSH-Px)等指标进行检测。结果:雷公藤内酯醇100 g·L-1组可明显抑制肝细胞的增殖,降低肝细胞活性,使细胞ALT,AST,LDH的活性升高,MDA含量升高,SOD和GSH-Px活性降低。结论:雷公藤内酯醇对肝细胞具有明显的损伤作用且随剂量增加具有一定的剂量依赖性,作用机制与其所致细胞脂质过氧化有关。
OBJECTIVE: To establish a method of high density culture of LO2 hepatocyte cell line using microcarriers to investigate the hepatotoxicity and mechanism of triptolide. METHODS: Cytodex-3 was used as a material to culture LO2 hepatocytes in vitro. The growth of LO2 hepatocytes was observed under light microscope. The effects of triptolide on the proliferation of hepatocytes were investigated. The alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) Glutathione peroxidase (GSH-Px) and other indicators were detected. Results: Triptolide 100 g · L-1 significantly inhibited the proliferation of hepatocytes and decreased the activity of hepatocytes. The activities of ALT, AST and LDH were increased, MDA content and SOD and GSH-Px activities were decreased . CONCLUSION: Triptolide has a significant damage on hepatocytes and dose-dependently with increasing dosage, and its mechanism may be related to lipid peroxidation induced by triptolide.