Harmol hydrochloride dihydrate induces autophagy in neuro cells and promotes the degradation of α-Syn by Atg5/Atg12-dependent pathway,Food Science & Nutrition

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Harmol hydrochloride dihydrate induces autophagy in neuro cells and promotes the degradation of α-Syn by Atg5/Atg12-dependent pathway
Food Science & Nutrition ( IF 3.5 ) Pub Date : 2022-08-12 , DOI: 10.1002/fsn3.3031
Gulinuer Abulimiti ₁ , Jianghua Zeng ₁ , Mutalifu Aimaiti ₂ , Xiuying Lei ₃ , Na Mi ₄

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Harmol hydrochloride dihydrate (HHD) is a novel alkaloid salt of the natural β-carboline harmol, which is isolated from Peganum harmala L. Here, we studied whether HHD could induce autophagy in neuro cells and investigated the underlying molecular mechanism. After incubation with HHD, the number of GFP-LC3 puncta in cells was measured using confocal microscopy. The distribution and colocalization of autophagosomes and autolysosomes in the cells were also detected. LC3 was gathered and cultured in a medium containing HHD. Compared with control cells and cells starved for 2 h, the number of GFP-LC3 puncta and the LC3-II expression level were significantly increased in HHD-treated cells (p < .05). The number of autophagosome (red) was increased and most of them were colocalized with lysosomes (green). Moreover, HHD induced the formation of puncta with Lysotracker Red positive in the L3 fat bodies (p < .05). When treated HEK cells with HHD, the protein expression level of LC3-II was markedly increased, and the protein expression level of α-Syn was significantly decreased (p < .05). HHD could induce the increased autophagosome in neuro cells by induction of autophagy. Moreover, HHD may promote the degradation of α-Syn protein to protect neuro cells by inducing autophagy.

中文翻译：

Harmol 盐酸盐二水合物诱导神经细胞自噬并通过 Atg5/Atg12 依赖性途径促进 α-Syn 降解

Harmol 盐酸盐二水合物 (HHD) 是天然 β-咔啉 harmol 的一种新型生物碱盐，从骆驼蓬中分离出来。在此，我们研究了 HHD 是否可以诱导神经细胞自噬，并探讨其潜在的分子机制。与 HHD 孵育后，使用共聚焦显微镜测量细胞中 GFP-LC3 斑点的数量。还检测了自噬体和自溶酶体在细胞中的分布和共定位。收集LC3并在含有HHD的培养基中培养。与对照细胞和饥饿 2 小时的细胞相比，HHD 处理的细胞中 GFP-LC3 斑点的数量和 LC3-II 表达水平显着增加 ( p < .05)。自噬体（红色）的数量增加，其中大部分与溶酶体（绿色）共存。此外，HHD 诱导 L3 脂肪体中 Lysotracker Red 呈阳性的斑点形成 ( p < .05)。当用HHD处理HEK细胞时，LC3-II的蛋白表达水平显着增加，而α-Syn的蛋白表达水平显着降低( p < .05)。 HHD可以通过诱导自噬来诱导神经细胞中自噬体的增加。此外，HHD可能促进α-Syn蛋白的降解，通过诱导自噬来保护神经细胞。

更新日期：2022-08-12

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