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槲皮素减轻6-羟基多巴胺诱导的SH-SY5Y细胞凋亡。

Quercetin alleviates 6-OHDA-caused apoptosis in SH-SY5Y cells.

作者信息

Pakrashi Sourav, Chakraborty Joyeeta, Bandyopadhyay Jaya

机构信息

Department of Biotechnology, Maulana Abul Kalam Azad University of Technology, NH-34 Road, District, Nadia, Haringhata, West Bengal 741249, India.

Department of Microbiology, Bidhannagar College, Block - EB2, Sector 1, Salt Lake, District - 24 Parganas (North), West Bengal 700064, India.

出版信息

Toxicol Res (Camb). 2024 Aug 2;13(4):tfae117. doi: 10.1093/toxres/tfae117. eCollection 2024 Aug.

Abstract

AIM

This study is primarily designed to investigate the potential neuroprotective effects of polyphenol against 6-OHDAcaused neurotoxicity on SH-SY5Y cells.

MATERIALS AND METHODS

Cytotoxic effect of 6-OHDA and valuable role of quercetin, myricetin and kaempferol on SH-SY5Y cells were analyzed by MTT assay. Generation of 6-OHDA-stimulated reactive oxygen species (ROS) was measured using DCFDA fluorescence dye. Alteration of 6-OHDA-caused mitochondrial membrane potential and nuclear condensation was investigated with the help of rhodamine-123 and hoechest stain. Immunoblotting was performed to detect the expression level of 6-OHDA-caused alpha-synuclein (á-syn), Bcl-2 associated protein X (BAX), caspase 3, cleaved Poly ADP - ribose polymerase (PARP) and Bcell lymphoma 2 proteins (Bcl-2).

RESULT

Through MTT assay, quercetin was selected over myricetin and kaempferol to counter 6-OHDA-caused cell death. The research delves into unraveling the intricate mechanisms underlying 6-OHDA-induced neurotoxicity, encompassing alterations in cellular morphology, escalation of oxidative stress, perturbation in mitochondrial membrane potential, and nuclear condensation. Exposure to 6-OHDA is implicated in the upregulation of á-syn protein, contributing to the aggravation of neurotoxicity. Concurrently, 6-OHDA orchestrates the apoptotic pathway by upregulating the expression of proapoptotic proteins such as BAX, caspase 3, and PARP, while down regulating the expression of the Bcl-2, affirming its role in apoptosis induction. Quercetin demonstrated ability to attenuate the expression of á-syn in the presence of 6-OHDA-caused injury in SH-SY5Y cells.

CONCLUSION

Taken together, these findings collectively underscore the therapeutic potential of quercetin as a promising agent against neurotoxicity caused by 6-OHDA.

摘要

目的

本研究主要旨在探讨多酚对6-羟基多巴胺(6-OHDA)引起的SH-SY5Y细胞神经毒性的潜在神经保护作用。

材料与方法

采用MTT法分析6-OHDA的细胞毒性以及槲皮素、杨梅素和山奈酚对SH-SY5Y细胞的重要作用。使用DCFDA荧光染料测量6-OHDA刺激产生的活性氧(ROS)。借助罗丹明-123和 Hoechst 染色研究6-OHDA引起的线粒体膜电位变化和核浓缩。进行免疫印迹以检测6-OHDA引起的α-突触核蛋白(α-syn)、Bcl-2相关蛋白X(BAX)、半胱天冬酶3、裂解的聚ADP-核糖聚合酶(PARP)和B细胞淋巴瘤2蛋白(Bcl-2)的表达水平。

结果

通过MTT法,选择槲皮素而非杨梅素和山奈酚来对抗6-OHDA引起的细胞死亡。该研究深入探究6-OHDA诱导神经毒性的复杂机制,包括细胞形态改变、氧化应激加剧、线粒体膜电位扰动和核浓缩。暴露于6-OHDA与α-syn蛋白上调有关,导致神经毒性加重。同时,6-OHDA通过上调促凋亡蛋白如BAX、半胱天冬酶3和PARP的表达来调控凋亡途径,同时下调Bcl-2的表达,证实其在诱导凋亡中的作用。槲皮素在6-OHDA对SH-SY5Y细胞造成损伤的情况下,表现出减弱α-syn表达的能力。

结论

综上所述,这些发现共同强调了槲皮素作为对抗6-OHDA引起的神经毒性的有前景药物的治疗潜力。

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