蛇菰宁通过抑制 p38 使 Neuro-2a/APP695 细胞免受氧化应激损伤。
Spinosin protects Neuro-2a/APP695 cells from oxidative stress damage by inactivating p38.
机构信息
School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang 110016, China.
School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
出版信息
J Tradit Chin Med. 2023 Oct;43(5):868-875. doi: 10.19852/j.cnki.jtcm.20220907.001.
OBJECTIVE
To explore the protective mechanism of spinosin (SPI) on Alzheimer's disease (AD) model cells, Neuro-2a/APP695 (N2a/APP695), against HO-induced oxidative stress damage, to reflect the influence of oxidative stress on the development of AD, and to provide a valuable basis for the research and development of therapeutic drug for AD.
METHODS
N2a/APP695 cells were exposed to HO and then treated with spinosin. Firstly, the secretion level of amyloid β (Aβ) and the production of malondialdehyde (MDA) and lactate dehydrogenase (LDH) were detected by enzyme linked immunosorbent assay kits. Secondly, the oligomerization degree of Aβ was performed by Thioflavin T staining. Thirdly, the expression levels of p-Tau (Ser199/202/396), synaptophysin (SYP), postsynaptic density protein 95 (PSD95), and mitogen-activated protein kinase (MAPK) family-related proteins were detected by Western blot analysis. In addition, FITC-labeled phalloidin was used in cytoskeleton staining to reflect synaptic function.
RESULTS
This study showed that HO stimulated N2a/APP695 cells to produce excessive MDA and LDH and secrete a large amount of Aβ, promoted the aggregation of Aβ, induced Tau protein hyperphosphorylation, and led to synaptic dysfunction. Spinosin reversed these changes caused by HO by inactivating p38, which was verified by treatment with the p38 inhibitor BIRB796.
CONCLUSION
Spinosin protects N2a/APP695 cells from oxidative stress damage caused by HO through inactivating p38.
目的
探讨桑枝生物碱（SPI）对阿尔茨海默病（AD）模型细胞Neuro-2a/APP695（N2a/APP695）氧化应激损伤的保护作用，反映氧化应激对 AD 发病的影响，为 AD 治疗药物的研究与开发提供有价值的依据。
方法
用 HO 处理 N2a/APP695 细胞，再用桑枝生物碱进行处理。首先，通过酶联免疫吸附试剂盒检测 Aβ的分泌水平、丙二醛（MDA）和乳酸脱氢酶（LDH）的产生。其次，通过 Thioflavin T 染色检测 Aβ的寡聚化程度。再次，通过 Western blot 分析检测 p-Tau（Ser199/202/396）、突触小体相关蛋白（SYP）、突触后密度蛋白 95（PSD95）和丝裂原活化蛋白激酶（MAPK）家族相关蛋白的表达水平。此外，用 FITC 标记的鬼笔环肽进行细胞骨架染色，以反映突触功能。
结果
本研究表明，HO 刺激 N2a/APP695 细胞产生过多的 MDA 和 LDH，并大量分泌 Aβ，促进 Aβ的聚集，诱导 Tau 蛋白过度磷酸化，导致突触功能障碍。桑枝生物碱通过激活 p38 来逆转 HO 引起的这些变化，这一作用可通过用 p38 抑制剂 BIRB796 处理来验证。
结论
桑枝生物碱通过抑制 p38 来保护 N2a/APP695 细胞免受 HO 引起的氧化应激损伤。
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