目的 研究蝦青素對七氟醚誘導的海馬神經元HT22細胞活力和細胞凋亡的作用與機制。方法 體外培養(yǎng)HT22細胞分為對照組、七氟醚組、七氟醚+蝦青素（1.25、2.50、5.00 μmol/L）組；七氟醚組給予4%七氟醚刺激細胞6 h；七氟醚+蝦青素組七氟醚刺激細胞前給予不同濃度的蝦青素處理2 h。采用MTT實驗檢測各組HT22細胞存活率，DCFH-DA熒光探針和硫代巴比妥酸法分別檢測各組細胞內的活性氧（ROS）水平和丙二醛（MDA）含量，流式細胞術和TUNEL檢測各組細胞凋亡，Western blotting檢測各組細胞中CyclinD1、Cleaved Caspase-3、Bax和Bcl-2、AMPK、p-AMPK、SIRT1蛋白的表達。結果 與對照組比較，七氟醚組HT22細胞存活率顯著下降和細胞凋亡率顯著增加（P<0.05），CyclinD1、Bcl-2、p-AMPK和SIRT1蛋白表達減少（P<0.05），Cleaved Caspase-3、Bax蛋白表達增加（P<0.05），ROS活性和MDA含量降低（P<0.05）。與模型組比較，七氟醚+蝦青素HT22細胞Cleaved Caspase-3、Bax蛋白表達下調（P<0.05），CyclinD1、Bcl-2、p-AMPK和SIRT1蛋白表達上調（P<0.05），ROS活性和MDA含量降低（P<0.05），且呈劑量相關性。結論 蝦青素對七氟醚誘導的HT22細胞凋亡和氧化應激損傷具有保護作用，其機制可能與調控AMPK-SIRT1通路有關。

Objective To study the effect and mechanism of astaxanthin on the viability and apoptosis of hippocampal neuron HT22 cells induced by sevoflurane. Methods HT22 cells cultured in vitro were divided into control group, sevoflurane group, sevoflurane + astaxanthin (1.25, 2.50, 5.00 μmol/L) group. The cells of sevoflurane group was given 4% sevoflurane stimulation and treated for 6 h; The cells of sevoflurane + astaxanthin group was treated with different concentrations of astaxanthin for 2 h before 4% sevoflurane stimulation. The MTT experiment was used to detect the survival rate of HT22 cells in each group, and the DCFH-DA fluorescent probe and thiobarbituric acid method were performed to determine the levels of ROS and MDA content. Flow cytometry and TUNEL were applied to detect cell apoptosis in each group, and Western blotting was used to detecte expression of CyclinD1, Cleaved caspase-3, Bax and Bcl-2, AMPK, p-AMPK and SIRT1 proteins. Results Compared with the control group, the survival rate of HT22 cells in the sevoflurane group was significantly decreased and the apoptosis rate was significantly increased. CyclinD1, Bcl-2, p-AMPK and SIRT1 protein expression were decreased, Cleaved Caspase-3, Bax protein expression were increased, and ROS activity and MDA content were decreased. Compared with the model group, the expression of Cleaved Caspase-3 and Bax protein were down-regulated in the astaxanthin group, and the expression of CyclinD1, Bcl-2, p-AMPK and SIRT1 protein were up-regulated, the activity of ROS and the content of MDA were decreased, which were dose-dependent. Conclusion Astaxanthin has a protective effect on sevoflurane induced HT22 cells apoptosis and oxidative stress injury, and its mechanism may be correlated to the regulation of AMPK-SIRT1 pathway.

R285

河南省醫(yī)學科技攻關計劃（2018020611）；河南省醫(yī)學科技攻關計劃（2018020653）；河南省醫(yī)學科技攻關計劃（2018020689）