川陈皮素抑制Aβ_(25~35)诱导的SH-SY5Y细胞氧化应激与凋亡作用

Nobiletin inhibits oxidative stress and apoptosis in Aβ_(25~35) induced SH-SY5Y cells

目的探讨川陈皮素对β淀粉样蛋白(Aβ)25~35诱导的SH-SY5Y细胞损伤的保护作用及潜在机制。方法SH-SY5Y细胞培养至对数生长期,分为正常对照组、Aβ_(25~35)损伤组及川陈皮素25、50、100μmol/L组,培养24 h后检测细胞存活率、氧化应激水平、凋亡率、凋亡相关基因及磷酸化-蛋白激酶B(p-Akt)、磷酸化-雷帕霉素靶蛋白(p-mTOR)蛋白表达等指标。结果与Aβ_(25~35)损伤组比较,川陈皮素各浓度处理组SH-SY5Y细胞的存活率显著增加(P<0.05),而乳酸脱氢酶(LDH)活性显著降低(P<0.05);与Aβ_(25~35)损伤组比较,川陈皮素各浓度处理组SH-SY5Y细胞谷胱甘肽过氧化酶(GSH-Px)、过氧化氢酶(CAT)活性显著增加(P<0.05),而丙二醛(MDA)含量显著降低(P<0.05);给予不同浓度川陈皮素处理后,与Aβ_(25~35)损伤组比较,SH-SY5Y细胞凋亡率、Bax mRNA表达显著降低(P<0.05),而Bcl-2 mRNA、Bcl-2/Bax值、p-Akt及p-mTOR蛋白表达显著增加(P<0.05)。结论川陈皮素可以通过调控Akt/mTOR通路抑制Aβ_(25~35)诱导的SH-SY5Y细胞氧化应激及凋亡,进而对Aβ_(25~35)诱导的SH-SY5Y细胞损伤产生保护作用。

Objective To investigate the protective effect and potential mechanism of nobiletin on injury of Aβ_(25~35) induced SH-SY5Y cells.Methods SH-SY5Y cells were cultured to logarithmic growth stage and divided into normal control group,Aβ_(25~35) injury group and 25,50 and 100μmol/L nobiletin groups.After 24 h of culture,cell survival rate,oxidative stress level,apoptosis rate,apoptosis related genes and p-Akt,p-mTOR protein were detected.Results Compared with the Aβ_(25~35) injury group,the survival rate of SH-SY5Y cells was increased(P<0.05)and the LDH activity was decreased(P<0.05)in the treatment group with different concentrations of nobiletin.Compared with the Aβ_(25~35) injury group,the activities of GSH-Px and CAT in SH-SY5Y cells were increased(P<0.05),while the MDA content was decreased(P<0.05)in the each dose nobiletin groups.Compared with those of the Aβ_(25~35) injury group,after the treatment with different concentrations of the nobiletin,the apoptosis rate and Bax mRNA expression of SH-SY5Y cells were decreased(P<0.05),while the expressions of Bcl-2 mRNA,Bcl-2/Bax ratio,p-Akt and p-mTOR protein were increased(P<0.05).Conclusions Nobiletin could inhibit the oxidative stress and apoptosis in Aβ_(25~35) induced SH-SY5Y cells through the regulation of Akt/mTOR pathway,thus protecting against injury of Aβ_(25~35) induced SH-SY5Y cells.