[1]王虎清,樊嘉欣,陈婉莹,等.PPARγ通路激活可增强细胞抗氧化能力和保护长程培养原代神经细胞[J].南方医科大学学报,2019,(01):23.[doi:10.12122/j.issn.1673-4254.2019.01.04]
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PPARγ通路激活可增强细胞抗氧化能力和保护长程培养原代神经细胞()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2019年01期
页码:
23
栏目:
出版日期:
2019-01-26

文章信息/Info

Title:
Activation of PPARγ pathway enhances cellular anti-oxidant capacity to protect long-term cultured primary rat neural cells from apoptosis
作者:
王虎清樊嘉欣陈婉莹高震张桂莲吴海琴俞小瑞
关键词:
过氧化物酶体增殖物激活受体原代神经细胞凋亡抗氧化
Keywords:
peroxisome proliferator activated receptor γ primary nerve cells aging anti-oxidation
DOI:
10.12122/j.issn.1673-4254.2019.01.04
摘要:
目的研究PPARγ通路增强对长程培养原代神经细胞凋亡的保护作用。方法以长程培养22 d的原代神经细胞建立自然 衰老模型。根据不同干预方法分为衰老对照组(C组)、GW9662 干预组(G组)和吡格列酮(Pioglitazone)干预组(P组)。其中 GW9662干预组(G组)根据GW9662干预终浓度分为4个亚组:G(0.1):0.1 μmol/Lol/L,G(1):1 μmol/L,G(5)5 μmol/L,G(10): 10 μmol/L。吡格列酮干预组(P组):根据吡格列酮干预终浓度分为4个亚组:P(0.1):0.1 μmol/L,P(1):1 μmol/L,P(5)5 μmol/L, P(10):10 μmol/L。首先应用MTT法观察各组细胞活力,应用倒置显微镜观察各组神经细胞形态,确定下一步实验的G组和P 组的最终干预浓度。其次应用免疫荧光双标染色、流式细胞仪观察各组神经细胞计数、凋亡细胞比率的变化,明确PPARγ通路 增强对长程培养原代神经细胞凋亡的保护作用。最后应用免疫化学法观察各组细胞抗氧化能力的变化,探讨PPARγ通路增强 保护原代神经细胞的机制。结果随着GW9662干预浓度的增加,细胞活力进行性下降,其中G(1)、G(5)和G(10)组细胞活力 均较C组显著降低(P<0.05)。G(1)组神经细胞形态尚完整,部分突触交织成网,最终确定后续实验G组的GW9662干预浓度为 1 μmol/L。加用吡格列酮干预后细胞活力较C组提高,其中P(1)、P(5)和P(10)组细胞活力较C组显著提高,差异具有统计学意 义(P<0.05),但P(1)、P(5)、P(10)组3组之间比较无明显差异(P>0.05),最终确定后续实验P组的吡格列酮干预浓度为1 μmol/L。 G组细胞总数及神经细胞计数均较C组显著下降(P<0.05)。P组细胞总数及神经细胞计数均高于C组(P<0.05)。各组神经细 胞比率均维持在80%左右,差异无统计学意义(P>0.05)。G组凋亡细胞比率明显高于C组(P<0.05),而P组凋亡细胞比率显著 低于C组(P<0.05)。G组SOD活性及GSH含量较C组均显著降低(P<0.05),而MDA含量显著增加(P<0.05)。P组SOD活性 及GSH含量均较C组显著升高(P<0.05),而MDA含量较C组显著减少(P<0.05)。结论PPARγ通路是长程培养原代神经细胞 的保护性通路,该通路的激活可以增强细胞抗氧化能力减少细胞凋亡保护长程培养原代神经细胞,提示针对PPARγ通路进行的 药物干预有可能是临床抗神经系统衰老治疗的有效途径之一。
Abstract:
Objective To study the protective effect of enhanced peroxisome proliferator activated receptor γ (PPARγ) pathway against apoptosis of long-term cultured primary nerve cells. Methods A natural aging model was established in primary rat nerve cells by long-term culture for 22 days. The cells were divided into control group, 0.1, 1.0, 5.0, and 10 μmol/L GW9662 intervention groups, and 0.1, 1.0, 5.0, and 10 μmol/L pioglitazone intervention groups. The cell viability was assessed using MTT assay and the cell morphological changes were observed after the treatments to determine the optimal concentrations of GW9662 and pioglitazone. Double immunofluorescence labeling and flow cytometry were used to observe the changes in the number of viable cells and cell apoptosis following the treatments; immunocytochemical staining was used to assess the changes in the anti-oxidation ability of the treated cells. Results The optimal concentrations of GW9662 and pioglitazone determined based on the cell viability and morphological changes were both 1 μmol/L. Compared with the control group, GW9662 treatment significantly lowered while pioglitazone significantly increased the total cell number and nerve cell counts (P<0.05), and nerve cells in the cell cultures maintained a constant ratio at about 80% in all the groups (P>0.05). GW9662 significantly enhanced while pioglitazone significantly lowered the cell apoptosis rates compared with the control group (P< 0.05). GW9662 obviously lowered SOD activity and GSH content in G group (P<0.05) and increased MDA content in the cells (P<0.05), and pioglitazone resulted in reverse changes in SOD, GSH and MDA contents in the cells (P<0.05). Conclusion Activation of PPARγ pathway protects long-term cultured primary nerve cells by enhancing cellular anti-oxidant capacity and reducing cell apoptosis, suggesting a potential strategy for anti-aging treatment of the nervous system through intervention of the PPARγ pathway.
更新日期/Last Update: 1900-01-01