【摘要】 目的建立猪肾近曲小管上皮细胞(LLC-PK1)的高糖损伤模型,观察α-亚麻酸(ALA)对高糖损伤LLC-PK1细胞的保护作用并探讨其作用机制。方法 CCK-8试剂盒测定葡萄糖对LLC-PK1细胞增殖的影响,流式细胞术测定不同浓度ALA干预高糖损伤LLC-PK1的凋亡率和活性氧(ROS)含量。结果高糖环境可以抑制体外培养的LLC-PK1细胞的增殖,形成体外高糖损伤模型;经适当浓度(50～100μmol/L)的ALA干预后,前干预组和持续干预组细胞的凋亡率显著低于阳性对照组(P<0.05);当ALA浓度为10～100μmol/L时,持续干预组LLC-PK1细胞内ROS含量显著低于阳性对照组(P<0.05),当ALA浓度为50μmol/L时,前干预组LLC-PK1细胞内ROS含量显著低于阳性对照组(P<0.05)。结论高糖损伤LLC-PK1模型为研究DN肾小管上皮细胞的防治干预提供了良好的体外研究平台,ALA有望成为预防肾小管高糖损伤的保护剂,减少活性氧的产生可能是ALA保护肾小管上皮细胞的作用机制之一。更多还原

【Abstract】 Objective Made LLC-PK1 damage model induced by high glucose and observing the protection effect and its mechanisms of LLC-PK1 injury induced by high glucose.Methods The proliferation of LLC-PK1 induced by high glucose was tested by CCK-8 and the apoptosis rat and the contents of reactive oxygen species(ROS) of LLC-PK1 damaged by high glucose was observed by flow cytometry after administration of different concentration α-linolenic acid(ALA).Result High glucose could obviously inhibit the proliferation of LLC-PK1.The apoptotic rates of LLC-PK1 intervened by ALA(50~100μmol/L) in the preconditioning group and the persistent intervention group were lower than those in the positive control group(P<0.05).The contents of ROS of LLC-PK1 in the persistent intervention group were lower than those in the positive control group when the concentration of ALA were from 10μmol/L to 100μmol/L(P <0.05,P <0.01).The contents of ROS of LLC-PK1 in the preconditioning group were lower than those in the positive control group when the concentration of ALA was 50μmol/L(P <0.05).Conclusion The model of LLC-PK1 induced by high glucose provided fine chances for the intervention of renal tubular epithelial cells in DN.ALA were expected to be a protectant to prevent high glucose damage of renal tubulars.Decreasing the active oxygen generation may be one of the mechanism of the protective effects on LLC-PK1 by ALA.更多还原