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李晓凤, 田梦芝, 陈笑一, 彭南波, 陈思远, 杜可.姜黄素通过Akt/mTOR/HIF-1α通路调控氧糖剥夺PC12细胞自噬的机制研究[J].湖南中医药大学学报英文版,2025,45(5):836-844.[Click to copy
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| 姜黄素通过Akt/mTOR/HIF-1α通路调控氧糖剥夺PC12细胞自噬的机制研究 |
| 李晓凤,田梦芝,陈笑一,彭南波,陈思远,杜可 |
| (湖南中医药大学, 湖南 长沙 410208;长沙市中心医院, 湖南 长沙 410004) |
| 摘要: |
| 目的 探讨姜黄素对氧糖剥夺（OGD）诱导的PC12细胞自噬损伤的保护作用及其机制。方法 通过体外建立OGD诱导的PC12细胞自噬损伤模型，将PC12细胞分为CON组、OGD组、右美托咪啶（DEX）组和姜黄素低、中、高剂量组。除CON组外，其余各组均换DMEM无糖培养基，并且在94%N2、5%CO2、1%O2条件下培养 6 h进行OGD处理。姜黄素低、中、高剂量组和DEX组分别加入1.25、5、20 μmol/L浓度的姜黄素以及0.5 μmol/L的DEX干预12 h。采用MTT法检测PC12细胞存活率；透射电子显微镜观察姜黄素处理后的自噬小体及自噬溶酶体；MDC荧光染色法观察自噬小体的荧光强度；Western blot检测微管相关蛋白轻链3（LC3）-Ⅱ/LC3-Ⅰ、苄氯素1（Beclin-1）、螯合体1（p62）、磷酸化蛋白激酶B/蛋白激酶B（p-Akt/Akt）、磷酸化雷帕霉素/雷帕霉素（p-mTOR/mTOR）、缺氧诱导因子1α（HIF-1α）等自噬相关蛋白表达情况。结果 与CON组相比，OGD组细胞存活率显著降低（P＜0.01），PC12细胞自噬小体荧光强度显著增强（P＜0.01）；LC3-Ⅱ/LC3-I、Beclin-1、HIF-1α蛋白表达显著升高（P＜0.01），p62、p-Akt/Akt、p-mTOR/mTOR蛋白表达显著降低（P＜0.01）。与OGD组相比，姜黄素低、中、高剂量组及DEX组细胞存活率均升高（P＜0.05，P＜0.01）；PC12细胞自噬小体荧光强度减弱（P＜0.05，P＜0.01）。与OGD组相比，姜黄素低、中剂量组及DEX组LC3-Ⅱ/LC3-Ⅰ、Beclin-1、HIF-1α蛋白表达显著降低（P＜0.01），p62、p-Akt/Akt、p-mTOR/mTOR蛋白表达显著升高（P＜0.01）。结论 姜黄素的低、中剂量组可通过激活Akt/mTOR通路，继而影响HIF-1α蛋白表达，抑制OGD诱导的PC12细胞自噬损伤，从而发挥抗神经元自噬损伤的保护作用。 |
| 关键词: 姜黄素 氧糖剥夺 细胞自噬 右美托咪啶 Akt/mTOR/HIF-α通路 自噬损伤 |
| DOI：10.3969/j.issn.1674-070X.2025.05.008 |
| Received:March 04, 2025 |
| 基金项目:湖南省自然科学基金项目（2025JJ80168）；湖南省普通高等学校教学改革研究项目（HNJG-2022-0134）；湖南中医药大学学科建设“揭榜挂帅”项目（22JBZ020）。 |
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| Mechanism of curcumin in regulating autophagy in oxygen-glucose deprived PC12 cells through the Akt/mTOR/HIF-1α pathway |
| LI Xiaofeng, TIAN Mengzhi, CHEN Xiaoyi, PENG Nanbo, CHEN Siyuan, DU Ke |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Changsha Central Hospital, Changsha, Hunan 410004, China) |
| Abstract: |
| Objective To explore the protective effects of curcumin on oxygen-glucose deprivation (OGD)-induced autophagic injury in PC12 cells and its mechanism. Methods An in vitro OGD-induced autophagic injury model was established in PC12 cells, and the PC12 cells were divided into CON group, OGD group, dexmedetomidine (DEX) group, and low-, medium-, and high-dose curcumin groups. Except for the CON group, cells in all other groups were switched to glucose-free DMEM medium and subsequently subjected to OGD treatment by incubating them for six hours under conditions of 94% N2, 5% CO2, and 1% O2. The low-, medium-, and high-dose curcumin groups, along with the DEX group, were respectively intervened with curcumin at concentrations of 1.25, 5, and 20 μmol/L, as well as 0.5 μmol/L DEX for 12 hours. The viability of PC12 cells was measured using the MTT assay. Transmission electron microscope was used to observe autophagosomes and autolysosomes following curcumin treatment, and the MDC staining method was employed to observe the fluorescence intensity of autophagosomes. Western blot was conducted to determine the relative expression levels of autophagy-related proteins, including microtubule-associated protein light chain 3 (LC3)-II/LC3-I, Beclin-1, sequestosome 1 (p62), phosphorylated protein kinase B/protein kinase B (p-Akt/Akt), phosphorylated mammalian target of rapamycin/mammalian target of rapamycin (p-mTOR/mTOR), and hypoxia-inducible factor 1α (HIF-1α). Results Compared with the CON group, the OGD group exhibited a significantly reduced cell viability (P<0.01) and a markedly enhanced fluorescent intensity of autophagosomes in PC12 cells (P<0.01). The protein expression levels of LC3-II/LC3-I, Beclin-1, and HIF-1α were significantly upregulated (P<0.01), while those of p62, p-Akt/Akt, and p-mTOR/mTOR were significantly downregulated (P<0.01). Compared with the OGD group, the low-, medium-, and high-dose curcumin groups and the DEX group exhibited increased cell viability (P<0.05, P<0.01) and reduced autophagic fluorescence intensity in PC12 cells (P<0.05, P<0.01). Compared with the OGD group, the low- and medium-dose curcumin groups and the DEX group demonstrated significantly decreased protein expressions of LC3-II/LC3-I, Beclin-1, and HIF-1α (P<0.01), along with significantly elevated protein expressions of p62, p-Akt/Akt, and p-mTOR/mTOR (P<0.01). Conclusion The low- and medium-dose curcumin groups can exert protective effects against neuronal autophagic injury by activating the Akt/mTOR pathway, subsequently modulating HIF-1α protein expression and inhibiting OGD-induced autophagic injury in PC12 cells. |
| Key words: curcumin oxygen-glucose deprivation autophagy dexmedetomidine Akt/mTOR/HIF-α pathway autophagic injury |
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