School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong Province, 510006, PR China; Research Centre of Basic Intergrative Medicine, School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong Province, 510006, PR China; Guangzhou Huaxia Vocational College, Guangzhou, Guangdong Province, 510935, PR China.
School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong Province, 510006, PR China; Research Centre of Basic Intergrative Medicine, School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong Province, 510006, PR China.
J Ethnopharmacol. 2024 Mar 1;321:117292. doi: 10.1016/j.jep.2023.117292. Epub 2023 Oct 7.
Traditional Chinese medicine views kidney shortage as a significant contributor to the aetiology of Parkinson's disease (PD), a neurodegenerative condition that is closely linked to aging. In clinical, patients with Parkinson's disease are often treated with Testudinis Carapax et Plastrum (Plastrum Testudinis, PT), a traditional Chinese medication that tonifies the kidney. Previous research has demonstrated that ethyl stearate (PubChem CID: 8122), an active component of Plastrum Testudinis Extracted with ethyl acetate (PTE), may encourage neural stem cells (NSCs) development into dopaminergic (DAergic) neurons. However, the effectiveness and mechanism of cotransplantation of ethyl stearate and NSCs in treating PD model rats still require further investigation.
PD is a neurodegenerative condition marked by the loss and degradation of dopaminergic neurons in the substantia nigra of the midbrain. Synaptic damage is also a critical pathology in PD. Because of their self-renewal, minimal immunogenicity, and capacity to differentiate into dopaminergic (DAergic) neurons, NSCs are a prospective treatment option for Parkinson's disease cell transplantation therapy. However, encouraging transplanted NSCs to differentiate into dopaminergic neurons and enhancing synaptic plasticity in vivo remains a significant challenge in improving the efficacy of NSCs transplantation for PD. This investigation seeks to examine the efficacy of cotransplantation of NSCs and ethyl stearate in PD model rats and its mechanism related to synaptic plasticity.
On 6-hydroxydopamine-induced PD model rats, we performed NSCs transplantation therapy and cotransplantation therapy involving ethyl stearate and NSCs. Rotating behavior induced by apomorphine (APO) and pole climbing tests were used to evaluate behavioral changes. Using a variety of methods, including Western blotting (WB), immunofluorescence analysis, enzyme-linked immunosorbent assay, and quantitative real-time polymerase chain reaction (qRT-PCR), we examined the function and potential molecular mechanisms of ethyl stearate in combined NSCs transplantation therapy.
In the rat PD model, cotransplantation of ethyl stearate with NSCs dramatically reduced motor dysfunction, restored TH protein levels, and boosted dopamine levels in the striatum, according to our findings. Furthermore, the expression levels of SYN1 and PSD95, markers of synaptic plasticity, and BDNF, closely related to synaptic plasticity, were significantly increased. Cotransplantation with ethyl stearate and NSCs also increased the expression levels of Dopamine Receptor D1 (Drd1), an important receptor in the dopamine neural circuit, accompanied by an increase in MMP9 levels, ERK1/2 phosphorylation levels, and c-fos protein levels.
According to the results of our investigation, cotransplantation of ethyl stearate and NSCs significantly improves the condition of PD model rats. We found that cotransplantation of ethyl stearate and NSCs may promote the expression of MMP9 by regulating the Drd1-ERK-AP-1 pathway, thus improving synaptic plasticity after NSCs transplantation. These findings provide new experimental support for the treatment of PD with the kidney tonifying Chinese medicine Plastrum Testudinis and suggest a potential therapeutic strategy for PD based on cotransplantation therapy.
ETHNOPHARMACOLOGICAL 相关性：中医认为肾虚是帕金森病 (PD) 发病机制的重要因素，PD 是一种与衰老密切相关的神经退行性疾病。在临床中，帕金森病患者通常采用龟板（龟板，PT）治疗，这是一种补肾的中药。先前的研究表明，正十六烷酸乙酯（PubChem CID：8122）是从乙酸乙酯中提取的龟板的一种活性成分（PTE），可能会促进神经干细胞（NSCs）向多巴胺能（DAergic）神经元发育。然而，正十六烷酸乙酯和 NSCs 共移植治疗 PD 模型大鼠的效果和机制仍需要进一步研究。
PD 是一种神经退行性疾病，其特征是中脑黑质多巴胺能神经元的丧失和退化。突触损伤也是 PD 的一个关键病理学特征。由于神经干细胞具有自我更新、免疫原性低和分化为多巴胺能（DAergic）神经元的能力，因此它们是帕金森病细胞移植治疗的一种有前途的治疗选择。然而，促进移植的 NSCs 分化为多巴胺能神经元并增强体内突触可塑性仍然是提高 NSCs 移植治疗 PD 效果的一个重大挑战。本研究旨在研究 NSCs 和正十六烷酸乙酯共移植治疗 PD 模型大鼠的效果及其与突触可塑性相关的机制。
在 6-羟多巴胺诱导的 PD 模型大鼠上，我们进行了 NSCs 移植治疗和涉及正十六烷酸乙酯和 NSCs 的共移植治疗。使用阿朴吗啡（APO）诱导的旋转行为和杆攀爬测试来评估行为变化。通过 Western blot（WB）、免疫荧光分析、酶联免疫吸附测定和定量实时聚合酶链反应（qRT-PCR）等多种方法，我们研究了正十六烷酸乙酯在 NSCs 共移植治疗中的作用及其潜在的分子机制。
我们的研究结果表明，在大鼠 PD 模型中，正十六烷酸乙酯与 NSCs 的共移植显著降低了运动功能障碍，恢复了 TH 蛋白水平，并增加了纹状体中的多巴胺水平。此外，突触可塑性的标志物 SYN1 和 PSD95 以及与突触可塑性密切相关的脑源性神经营养因子（BDNF）的表达水平显著增加。正十六烷酸乙酯与 NSCs 的共移植还增加了多巴胺能神经回路中重要受体 Dopamine Receptor D1（Drd1）的表达水平，同时伴随着 MMP9 水平、ERK1/2 磷酸化水平和 c-fos 蛋白水平的升高。
根据我们的研究结果，正十六烷酸乙酯和 NSCs 的共移植显著改善了 PD 模型大鼠的状况。我们发现，正十六烷酸乙酯和 NSCs 的共移植可能通过调节 Drd1-ERK-AP-1 通路促进 MMP9 的表达，从而改善 NSCs 移植后的突触可塑性。这些发现为补肾中药龟板治疗 PD 提供了新的实验依据，并为基于共移植治疗的 PD 提供了一种潜在的治疗策略。
