红花黄色素对冈田酸诱导的人神经母细胞瘤SH-SY 5Y细胞Tau蛋白磷酸化的调节作用研究

马勤, 阮影影, 施晓萌, 王志祥, 胡艳丽

中国药学杂志 ›› 2016, Vol. 51 ›› Issue (3) : 213-217.

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中国药学杂志
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中国药学杂志 ›› 2016, Vol. 51 ›› Issue (3) : 213-217. DOI: 10.11669/cpj.2016.03.013
论 著

红花黄色素对冈田酸诱导的人神经母细胞瘤SH-SY 5Y细胞Tau蛋白磷酸化的调节作用研究

  • 马勤1a,2, 阮影影1a, 施晓萌1a, 王志祥1b, 胡艳丽1a*
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Study on Regulatory Effects of Safflower Yellow on Tau Protein Hyperphosphorylation in SH SY-5Y Cell Induced by Okadaic Acid

  • MA Qin1a,2, RUAN Ying-ying1a, SHI Xiao-meng1a, WANG Zhi-xiang1b, HU Yan-li1a*
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文章历史 +

摘要

目的 探讨红花黄色素(safflower yellow, SY)对冈田酸(okadaic acid, OA)诱导SH-SY 5Y细胞高磷酸化水平的调节作用及可能机制。方法 SH-SY 5Y细胞给予红花黄色素(0.01、0.1、1 g·L-1)处理,4 h后给予冈田酸 80 nmol·L-1处理48 h,利用MTT法测定红花黄色素对冈田酸损伤的SH-SY 5Y细胞存活率的影响。Hoechst 33342染色法观察红花黄色素对冈田酸诱导的SH-SY 5Y细胞损伤中凋亡的影响。Western blot检测红花黄色素对冈田酸诱导的SH-SY 5Y细胞中p-tau(T205、S199)、总GSK-3β 和PP2A的表达。结果 红花黄色素可明显增加冈田酸损伤的SH-SY 5Y细胞的生存率,Hoechst 33342染色显示红花黄色素可显著减少冈田酸损伤的SH-SY 5Y细胞的凋亡细胞个数。Western blot检测结果显示,p-tau(T205、S199)位点tau蛋白含量增加,红花黄色素能够显著降低tau蛋白的含量, 红花黄色素能够增加PP2A的表达和降低GSK-3β表达。结论 红花黄色素对冈田酸损伤的SH-SY 5Y细胞具有一定保护作用,并可能通过调节tau蛋白上游激酶和磷酸酶的蛋白表达而降低tau蛋白磷酸化水平。

Abstract

OBJECTIVE To investigate the regulating effects and its possible mechanism of safflower yellow (SY) on tau protein hyperphosphorylation in SH-SY 5Y cell induced by okadaic acid (OA). METHODS Cell viability was measured by MTT colorimetric method. The cell apoptosis was determined with Hoechst 33342 after SH-SY 5Y cell were treated with SY. To determined the phosphorylated tau protein of p-tau(T205), p-tau(S199) sites, and total GSK-3β, PP2A in SH-SY 5Y cell were determined using Western blotting. RESULTS SY significantly increased survival rates of SH-SY5Y cell damaged by OA. Hoechst 33342 showed that SY decreased the apoptosis of SH-SY5Y cells. Western blotting analysis showed that tau protein content was increased compared to control group at p-tau (T205), p-tau (S199) sites. However, SY treatment significantly reduced tau protein content at p-tau (T205) and p-tau (S199) sites. PP2A content was significantly suppressed by OA, significantly increased by SY, GSK-3β content was significantly increased by OA and significantly decreased by SY. CONCLUSION These results suggest that safflower yellow may have neuroprotective effect on OA-induced SH-SY 5Y cells injury, which may be associated with tau hyperphosphorylation at the enzyme level by activation of tau kinases or by a decline in the activities of tau phosphatases.

关键词

红花黄色素 / tau蛋白磷酸化 / 蛋白激酶 / 磷酸酶

Key words

safflower yellow / tau hyperphosphorylation / kinase / phosphatase

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马勤, 阮影影, 施晓萌, 王志祥, 胡艳丽. 红花黄色素对冈田酸诱导的人神经母细胞瘤SH-SY 5Y细胞Tau蛋白磷酸化的调节作用研究[J]. 中国药学杂志, 2016, 51(3): 213-217 https://doi.org/10.11669/cpj.2016.03.013
MA Qin, RUAN Ying-ying, SHI Xiao-meng, WANG Zhi-xiang, HU Yan-li. Study on Regulatory Effects of Safflower Yellow on Tau Protein Hyperphosphorylation in SH SY-5Y Cell Induced by Okadaic Acid[J]. Chinese Pharmaceutical Journal, 2016, 51(3): 213-217 https://doi.org/10.11669/cpj.2016.03.013
中图分类号： R965   

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基金

国家自然科学基金资助项目(81360495);兵团博士基金资助项目(2101701)

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