摘要目的 探讨蛋白激酶Cβ(protein kinase Cβ,PKCβ)抑制剂LY333531对氟诱导人神经母瘤细胞SH-SY5Y氧化损伤及凋亡的影响.方法 建立氟中毒SH-SY5Y细胞模型,分为对照组[0.0 mmol/L氟化钠(NaF)、0.0 μmol/L LY333531]、染氟组(0.5 mmol/L NaF、0.0 μmol/LLY333531)、PKC3抑制组(0.5 mmol/L NaF、0.2 μmol/L LY333531),n=3.流式细胞仪检测各组作用48 h后细胞内活性氧(reactive oxygen species,ROS)及细胞凋亡率,荧光探针法检测线粒体膜电位.结果 与对照组[(3.32±0.29)×103,0.60±0.09,(7.58±1.20)％]比较,染氟组ROS水平[(5.99±0.32)×103]升高,线粒体膜电位(0.28±0.06)降低,细胞凋亡率[(18.00±2.32)％]增加(P均＜0.05);与染氟组比较,PKCβ抑制组ROS水平[(5.12±0.25)×103]降低,线粒体膜电位(0.42±0.03)升高,细胞凋亡率[(11.79±0.70)％]减少(P均＜0.05).结论 过量氟会导致细胞发生氧化损伤及凋亡,而PKC3抑制剂LY333531可能对氟诱导的细胞氧化损伤及凋亡有一定的保护作用.

abstractsObjective To explore the influences of protein kinase Cβ (PKC3) inhibitor LY333531 on oxidative injury and apoptosis of SH-SY5Y cells induced by fluorosis.Methods The SH-SY5Y cell model of fluorosis was established,and the experiment was divided into three groups:control group [0.0 mmol/L sodium fluoride (NaF) and 0.0 μmol/L LY333531],the fluoride group (0.5 mmol/L NaF and 0.0 μmol/L LY333531),and the PKCβ inhibitor group (0.5 mmol/L NaF and 0.2 μmol/L LY333531),n =3.Flow cytometry was used to detect the changes of reactive oxygen species (ROS) and apoptosis rate,fluorescent probe technique was used to detect mitochondrial membrane potential after each group for 48 h.Results Compared with the control group [(3.32 ± 0.29) × 103,0.60 ± 0.09,(7.58 ± 1.20)％],the level of ROS [(5.99 ± 0.32) × 103] was increased,mitochondrial membrane potential (0.28 ± 0.06) was decreased,and the apoptosis rate [(18.00 ± 2.32)％] was increased in the fluoride group (all P ＜ 0.05);compared with the fluoride group,the level of ROS [(5.12 ± 0.25) × 103] was decreased,mitochondrial membrane potential (0.42 ± 0.03) was increased,and the apoptosis rate [(11.79 ± 0.70)％] was decreased in the PKCβ inhibitor group (all P ＜ 0.05).Conclusions Excess fluoride could cause oxidative damage and apoptosis in cells.PKC3 inhibitor LY333531 has a protective effect in oxidative damage and apoptosis by fluorosis.