Department of Pharmacology, Qingdao University School of Pharmacy, Qingdao, Shandong 266021, P.R. China.
School of Basic Medical Sciences, Qingdao University, Qingdao, Shandong 266021, P.R. China.
Mol Med Rep. 2017 Dec;16(6):8721-8728. doi: 10.3892/mmr.2017.7725. Epub 2017 Oct 4.
Accumulation of α‑synuclein (α‑SYN) is a common pathology for Parkinson's disease (PD). There is abundant evidence that the toxic‑gain‑of‑function of α‑SYN's is associated with aggregation and consequent effects. To assess the potential of chondroitin sulfate (CS) in this regard, the present study investigated its neuroprotective on SH‑SY5Y cells overexpressing wild‑type (WT) or A53T mutant α‑SYN. Cell viability was measured by MTT assay. Apoptosis, reactive oxygen species (ROS) and mitochondrial membrane potential were detected by flow cytometry. The protein expression levels of total α‑SYN, phosphorylated Ser129 α‑SYN, B‑cell lymphoma 2 (Bcl‑2), Bcl‑2‑associated X protein (Bax) and cytochrome‑c (Cyt‑c ) were analyzed by western blotting. It was observed that CS reduced the expression levels of total α‑SYN and phosphorylated Ser129 α‑SYN, prevented cell loss and inhibited apoptosis. The subsequent mechanism study indicated that CS inhibited ROS overproduction. CS also significantly attenuated WT and A53T mutant α‑SYN‑induced dysfunction, including decrease of mitochondrial membrane potential, decrease of Bcl‑2 expression, and increase of Bax expression, release of Cyt‑c from the mitochondria and activation of caspase‑3 and caspase‑9, which demonstrated that CS suppressed α‑SYN‑induced apoptosis possibly through mitochondria protection. These results suggested that CS protects SH‑SY5Y cells overexpressing WT or A53T mutant α‑SYN by inhibiting the expression and phosphorylation of α‑SYN, and ROS overproduction and mitochondrial apoptosis. These results implicate CS as a potential therapeutic agent for the treatment of PD.
α-突触核蛋白(α-SYN)的积累是帕金森病(PD)的常见病理。有大量证据表明,α-SYN 的毒性获得功能与其聚集和随之而来的影响有关。为了评估硫酸软骨素(CS)在这方面的潜力,本研究调查了 CS 对过表达野生型(WT)或 A53T 突变型 α-SYN 的 SH-SY5Y 细胞的神经保护作用。通过 MTT 测定法测量细胞活力。通过流式细胞术检测细胞凋亡、活性氧(ROS)和线粒体膜电位。通过蛋白质印迹法分析总 α-SYN、磷酸化 Ser129 α-SYN、B 细胞淋巴瘤 2(Bcl-2)、Bcl-2 相关 X 蛋白(Bax)和细胞色素 c(Cyt-c)的蛋白表达水平。结果观察到 CS 降低了总 α-SYN 和磷酸化 Ser129 α-SYN 的表达水平,防止了细胞丢失并抑制了细胞凋亡。随后的机制研究表明 CS 抑制了 ROS 的过度产生。CS 还显著减轻了 WT 和 A53T 突变型 α-SYN 诱导的功能障碍,包括线粒体膜电位降低、Bcl-2 表达降低、Bax 表达增加、Cyt-c 从线粒体释放以及 caspase-3 和 caspase-9 的激活,表明 CS 通过抑制线粒体凋亡来抑制 α-SYN 诱导的细胞凋亡。这些结果表明 CS 通过抑制 α-SYN 的表达和磷酸化、ROS 的过度产生和线粒体凋亡来保护过表达 WT 或 A53T 突变型 α-SYN 的 SH-SY5Y 细胞。这些结果表明 CS 可能是治疗 PD 的潜在治疗剂。