摘要
目的探讨异鼠李素(ISO)对高糖高脂诱导小鼠胰岛β细胞株MIN6细胞损伤的保护作用及机制。方法不同浓度ISO预处理MIN6细胞后高糖高脂培养48 h,CCK8法筛选ISO最佳干预浓度。细胞分为正常对照(Con)组(11.1 mmol/L葡萄糖)、高糖高脂损伤模型(M)组(33.3 mmol/L葡萄糖+0.25 mmol/L棕榈酸)及ISO预处理(ISO+M)组(10μmol/L ISO+33.3 mmol/L葡萄糖+0.25 mmol/L棕榈酸)。流式细胞术测定各组细胞凋亡率;硝酸还原酶法检测一氧化氮(NO)含量;RT-PCR测定IL-1β、IL-6、肿瘤坏死因子α(TNF-α)mRNA表达;Western blot法检测磷酸化核因子κB(NF-κB)抑制蛋白(IκB)激酶β(p-IKKβ)、磷酸化NF-κB抑制蛋白α(p-IκBα)、诱导型一氧化氮合酶(iNOS)及NF-κB p65蛋白的表达。结果细胞凋亡及NO含量M组高于Con组,ISO+M组低于M组(P<0.01)。IL-1β、IL-6、TNF-αmRNA,p-IKKβ、p-IκB、iNOS蛋白与NF-κB p65蛋白表达M组高于Con组,ISO+M组低于M组(P<0.05)。结论ISO可减轻高糖高脂诱导的胰岛β细胞损伤,可能与抑制IκB激酶(IKK)/IκB/NF-κB/iNOS通路活性有关。
Objective To investigate the protective effect and mechanism of Isorhamnetin(ISO)on the injury of pancreatic beta cells induced by high glucose and fatty acid.treated with various concentration of ISO under high glucose and fatty acid conditions for 48 h to select the optimum ISO intervention concentration by CCK8 method.Cells were divided into three groups:normal control(Con)group(11.1 mmol/L glucose),model(M)group(33.3 mmol/L glucose+0.25 mmol/L palmitic acid),ISO+M group(10μmol/L ISO+33.3 mmol/L glucose+0.25 mmol/L palmitic acid).Cell apoptosis was analyzed by flow cytometry.Nitric oxide(NO)content of cell supernatant was detected by nitrate reductase method.RT-PCR was used to detect the mRNA expression of interleukin 1β(IL-1β),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α).Western blot was used to determine intracellular protein expression of phosphorylated IκB kinaseβ(p-IKKβ),phosphorylated inhibitory kappa B(p-IκB),inducible nitric oxide synthase(iNOS)and the endonuclear protein expression of nuclear factor-kappa B(NF-κB).Results Cell apoptosis and NO content were higher in M group than in Con group and lower in ISO+M group than in M group(P<0.01).The mRNA expression of IL-1β,IL-6 and TNF-α,the protein expression of p-IKKβ,p-IκB and iNOS and the endonuclear protein expression of NF-κB p65 were higher in M group than in Con group and lower in ISO+M group than in M group(P<0.05).Conclusion ISO could alleviate the injury induced by glucolipotoxicity in islet beta cells,which may be related to the inhibition of IKK/IκB/NF-κB/iNOS pathway activity.
作者
来千惠
熊青
吴贵军
何光珍
徐焱成
LAI Qianhui;XIONG Qing;WU Guijun(Department of Endocrinology,Zhongnan Hospital of Wuhan University,Wuhan 430071,China)
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2020年第6期461-466,共6页
Chinese Journal of Diabetes
基金
国家自然科学基金(81370872)。
