PPARγ激动剂对抗经氟诱导的SH-SY5Y神经细胞氧化损伤
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摘要
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 15 d-PGJ2 on the oxidative damage induced by fluorosis in human neuroblastoma SH-SY5 Y cells. Methods: The SH-SY5 Y 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 15 d-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 correlation 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 PPARγ protein and SOD activity in the NaF group were significantly decreased,and the MDA content was significantly increased(P < 0. 05). In R +Na F group,the level of PPARγ protein expression and the activity of SOD in SH-SY5 Y 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-SY5 Y 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 15 dPGJ2 may attenuate the fluorine-induced cellular oxidative damage.
Objective: To observe the effect of peroxisome proliferator-activated receptor γ(PPARγ)agonist 15 d-PGJ2 on the oxidative damage induced by fluorosis in human neuroblastoma SH-SY5 Y cells. Methods: The SH-SY5 Y 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 15 d-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 correlation 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 PPARγ protein and SOD activity in the NaF group were significantly decreased,and the MDA content was significantly increased(P < 0. 05). In R +Na F group,the level of PPARγ protein expression and the activity of SOD in SH-SY5 Y 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-SY5 Y 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 15 dPGJ2 may attenuate the fluorine-induced cellular oxidative damage.
引文
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[17]YU X,SHAO X G,SUN H,et al. Activation of cerebral peroxisome proliferator-activated receptors gamma execs neuropro-tection by inhibiting oxidative stress following pilocarpine-inducedstatus epilepticus[J]. Brain Res,2008,20(8):146-158.
[18]邓永兵,唐文渊. PPAR的神经保护作用研究进展[J].中华神经疾病研究杂志,2010,9(1):92-94.
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[20]张婧媛,张艳桥,张一娜,等. PPARγ激动剂减轻缺氧性大鼠神经细胞损伤的作用机制[J].中国病理生理杂志,2009,25(1):89-92.
[2]GRAY E,GINTY M,KEMP K,et al. Peroxisome proliferator activated receptor-alpha agonists protect cortical neurons from inflammatory mediators and improve peroxisomal function[J]. Eur J Neurosci,2011,33(8):1421-1432.
[3]STORER P D,XU J,CHAVIS J,et al. Peroxisome proliferators-activated receptor-gamma agonists inhibit the activation of microglia and astrocytes:implications for multiple sclerosis[J]. J Neuroimmunol,2005,161(1-2):113-122.
[4]YU X,SHAO X G,SUN H,et al. Activation of cerebral peroxisome proliferator-activated receptors gamma exerts neuroprotection by inhibiting oxidative stress following pilocarpine-induced status epilepticus[J]. Brain Research,2008,1200(1):146-158.
[5]陈修文,万昌武,官志忠,等.氟中毒病区人群血清中多种氧化应激指标的综合分析[J].工业卫生与职业病,2016,42(4):249-252.
[6]官志忠,氧化应激在地方性氟中毒分子发病机制中的作用[J].中华地方病学杂志,2016,35(2):79-82.
[7]BARBIER O,ARREOLA-MENDOZA L,DEL RAZO L M. Molecular mechanisms of fluoride toxicit[J]. Chem Biol Interact,2010,188(2):319-333.
[8]刘燕斐,官志忠.慢性氟中毒性肝损伤及其发病机制[J].中国地方病学杂志,2012,31(5):588-590.
[9]VELLA S,CONALDI P G,FLORIO T,PAGANO A.PPAR gamma in neuroblastoma:the translational perspectives of hypoglycemic drugs[J]. PPAR Res, 2016,2016:3038164.
[10]CHOI S S,PARK J Y,CHOI J H. Revisiting PPARγas a target for the treatment of metabolic disorders[J]. BMB Rep,2014,47(11):599-608.
[11]CRAFT S,WASTON G S. Insulin and neurodegenerative disease:shared and specific mechanisms[J]. Lancet Neurol,2004,3(3):169-178.
[12]BLANQUICETT C,KANG B Y,RITZENTHALER J D,et al. Oxidative stress modulates PPAR gamma in vascular endothelial cells.[J]. Free Radical Biology&Medicine,2010,48(12):1618.
[13]MOHAPATRA S K,GURI A J,CLIMENT M,et al. Immunoregulatory actions of epithelial cell PPAR gamma at the colonic mucosa of mice with experimental inflammatory bowel disease.[J]. Plos One,2010,5(4):10215.
[14]LIU Z,LU G,CHEN Y,et al. Mark4 promotes oxidative stress and inflammation via binding to PPARgamma and activating NF-kappaB pathway in mice adipocytes[J].Scientific Reports,2016,6:21382.
[15]CHEN L,BI B,ZENG J,et al. Rosiglitazone ameliorates senescence-like phenotypes in a cellular photoaging model[J]. Journal of Dermatological Science,2015,77(3):173-181.
[16]KAPADIA R,YI J H,VEMUGANTI R. Mechanisms of anti-inflammatory and neuroprotective actions of PPARgamma agonists[J]. Front Biosci,2009,1(13):1813-1826.
[17]YU X,SHAO X G,SUN H,et al. Activation of cerebral peroxisome proliferator-activated receptors gamma execs neuropro-tection by inhibiting oxidative stress following pilocarpine-inducedstatus epilepticus[J]. Brain Res,2008,20(8):146-158.
[18]邓永兵,唐文渊. PPAR的神经保护作用研究进展[J].中华神经疾病研究杂志,2010,9(1):92-94.
[19]王宁,韩晶,徐艳炜,等.过氧化氢损伤原代皮质神经元中PPARγ的改变[J].中华医学杂志,2015,95(21):1686-1690.
[20]张婧媛,张艳桥,张一娜,等. PPARγ激动剂减轻缺氧性大鼠神经细胞损伤的作用机制[J].中国病理生理杂志,2009,25(1):89-92.