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目的:氧化应激在肝脏疾病中扮演着重要的角色。胶原蛋白肽是天然的抗氧化剂,其在动物实验中已经被证实有抑制氧化应激的作用。最新研究证实胶原蛋白肽将有可能被应用在肝脏疾病的预防中,但是很少有研究报道其分子作用机制。因此本研究在胶原蛋白肽是对H2O2诱导的正常人的肝细胞系HL7702氧化损伤有保护作用的基础上,并探索其分子作用机制。方法:实验设空白对照组,H2O2模型组,胶原蛋白肽低、中、高剂量组(10,100,200μg/ml)。胶原蛋白肽各组加入相应浓度的药物预处理12 h后,与模型组一起加入300μM H2O2的H2O2共同培养12 h,空白对照组正常培养。细胞毒性是由CCK8和乳酸脱氢酶(LDH)的释放检测。抗氧化试剂盒检测细胞内活性氧的水平,超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和丙二醛(MDA)含量的变化。Western blot检测细胞内Nrf2蛋白的表达水平。结果:胶原蛋白肽对H2O2诱导的正常人的肝细胞系HL7702氧化损伤有保护作用。胶原蛋白肽能够及时清除细胞内的活性氧,增加Nrf2的蛋白表达水平,提高超氧化物歧化酶(SOD)、过氧化氢酶(CAT)的活性,减轻脂质过氧化反应,从而保护正常人的肝细胞系HL7702。结论:总之,胶原蛋白肽通过增加Nrf2的蛋白表达水平,提高抗氧化活性,对H2O2诱导损伤的肝细胞发挥保护作用。本研究为胶原蛋白肽的分子作用机制提供了新的证据,将有助于预防氧化应激所致的肝损伤。
OBJECTIVE: Oxidative stress plays an important role in liver diseases. Collagen peptides are natural antioxidants that have been shown to inhibit oxidative stress in animal experiments. Recent research confirms that collagen peptides may be used in the prevention of liver diseases, but few studies have reported the molecular mechanism of action. Therefore, in this study, collagen peptide is a protective effect on oxidative damage of human hepatocyte cell line HL7702 induced by H2O2, and to explore its molecular mechanism. Methods: The experiment was set blank control group, H2O2 model group, collagen peptide low, medium and high dose group (10,100,200μg / ml). Collagen peptide groups were added into the corresponding concentration of drug pretreatment for 12 h, with the model group by adding 300 μM H2O2 H2O2 co-culture for 12 h, the blank control group normal culture. Cytotoxicity is detected by the release of CCK8 and lactate dehydrogenase (LDH). The level of reactive oxygen species (ROS), the activities of superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) Western blot detection of intracellular Nrf2 protein expression levels. RESULTS: Collagen peptide had a protective effect on oxidative damage of human hepatoma cell line HL7702 induced by H2O2. Collagen peptide can promptly remove intracellular reactive oxygen species, increase the protein level of Nrf2, increase the activity of superoxide dismutase (SOD) and catalase (CAT), reduce the lipid peroxidation and protect normal people Of hepatocyte cell line HL7702. Conclusion: In conclusion, collagen peptide can protect hepatocytes induced by H2O2 by increasing the protein level of Nrf2, enhancing the antioxidant activity. This study provides new evidence for the molecular mechanism of action of collagen peptides and will help prevent oxidative stress-induced liver injury.