PPARγ激动剂对抗经氟诱导的SH-SY5Y神经细胞氧化损伤

Study on PPARγ Agonist on SH-SY5Y Neuron Oxidative Damage Induced by Fluorosis

目的:观察过氧化物酶体增殖物激活受体γ(PPARγ)激动剂15d-PGJ2对氟诱导人神经母瘤SHSY5Y细胞氧化损伤的影响。方法:将体外培养的SH-SY5Y细胞,分为对照组、染氟组(Na F组)、单纯PPARγ激动剂组(R组)、干预组(R+Na F组,先加入15d-PGJ2 2 h后再加Na F),各组处理后培养48 h,用蛋白印记法检测SH-SY5Y细胞中PPARγ蛋白表达水平,微量酶标法测定超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量,分析PPARγ蛋白与SOD活性及MDA含量的相关关系。结果:与对照组相比,R组SH-SY5Y细胞中PPARγ蛋白水平、SOD活性及MDA含量无明显改变(P> 0. 05),Na F组SH-SY5Y细胞PPARγ蛋白水平和SOD活性明显降低,MDA含量明显增加(P <0. 05); R+Na F组SH-SY5Y细胞中PPARγ蛋白表达水平及SOD活性明显高于Na F组、MDA含量明显低于Na F组(P <0. 05); SH-SY5Y细胞中PPARγ蛋白表达水平与SOD活性呈正相关关系(r=0. 771,P <0. 05),与MDA含量呈负相关关系(r=-0. 762,P <0. 05)。结论:过量氟会导致体外培养的SH-SY5Y细胞发生氧化损伤,而PPARγ激动剂15d-PGJ2处理后可减轻氟诱导的细胞氧化损伤。

Objective： To observe the effect of peroxisome proliferator-activated receptor γ （PPARγ） agonist 15d-PGJ2 on the oxidative damage induced by fluorosis in human neuroblastoma SH-SY5Y cells. Methods： The SH-SY5Y cells were cultivated vitro. The cells were divided into normal control group, fluoride group （NaF group）, PPARγ agonist group （R group）, and intervention group （R＋ NaF group, cells were first treated with 20 hr 15d-PGJ2 first and then NaF） , the culture time of each group was 48 h. The level of PPARγ protein in the cells was detected by Western blotting. The activity of SOD and the content of MDA were determined by biochemical methods and the COlTelation between PPARγ protein, SOD activity and MDA content was analyzed. Result： Compared with the control group, there were no significant changes of the level of PPARγ protein, SOD activity, and MDA content in the R group （P 〉 0.05）, while the level of PPAR3, protein and SOD activity in the NaF group were significantly decreased, and the MDA content was significantly increased （P 〈 0.05）. In R ＋ NaF group, the level of PPARγ protein expression and the activity of SOD in SH-SY5Y cells were significantly higher, and the content of MDA was significantly lower than those in the fluoride group （P 〈 0.05 ）. The level of PPARγ protein in SH-SY5Y cells was positively correlated with SOD activity （r = 0.771, P 〈 0.05 ） and negatively correlated with MDA content （ r = - 0. 762, P 〈 0.05 ）. Conclusion ： Excess fluoride may cause oxidative damage to cultured neurons in vitro, whereas PPARγ agonist 15d- PGJ2 may attenuate the fluorine-induced cellular oxidative damage.