Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Pharmacology of Chinese Materia Medica, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198 China.
Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Resource and Developmemt of Chinese Material Medica, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198 China.
Phytomedicine. 2022 Jan;95:153882. doi: 10.1016/j.phymed.2021.153882. Epub 2021 Dec 28.
YiQiFuMai lyophilized injection (YQFM) is derived from a traditional Chinese medicine prescription termed Shengmai San.YQFM is clinically applied to the treatment of cardiovascular and cerebrovascular diseases. It has been found that critical components of YQFM affect non-muscle myosin heavy chain IIA (NMMHC IIA), but its regulation in the excessive autophagy and the underlying mechanism has yet to be clarified.
To evaluate whether YQFM has neuroprotective effects on cerebral ischemia/reperfusion-induced injury by inhibiting NMMHC IIA-actin-ATG9A interaction for autophagosome formation.
The neuroprotective effects of YQFM were investigated in vivo in mice with middle cerebral artery occlusion/reperfusion (MCAO/R) (n = 6) by detecting neurological deficits, infarct volume, and histopathological changes. The NMMHC IIA-actin-ATG9A interaction was determined using immunofluorescence co-localization, co-immunoprecipitation, and proximity ligation assay. Rat pheochromocytoma (PC12) cells subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) were used to mimic neurons in in vitro experiments.
In MCAO/R model mice, YQFM (1.342 g/kg) attenuated brain ischemia/reperfusion-induced injury by regulating NMMHC IIA-actin-mediated ATG9A trafficking. YQFM (400 μg/ml) also exerted similar effects on OGD/R-induced PC12 cells. Furthermore, RNAi of NMMHC IIA weakened the NMMHC IIA-F-actin-dependent ATG9A trafficking and, therefore, attenuated the neuroprotective activities of YQFM in vitro.
These findings demonstrated that YQFM exerted neuroprotective effects by regulating the NMMHC IIA-actin-ATG9A interaction for autophagosome formation. This evidence sheds new light on the potential mechanism of YQFM in the treatment of cerebral ischemia/reperfusion.
益气复脉冻干注射剂（YQFM）源自一种名为生脉散的中药方剂。YQFM 临床上用于治疗心脑血管疾病。现已发现，YQFM 的关键成分会影响非肌肉肌球蛋白重链 IIA（NMMHC IIA），但其在过度自噬中的调节作用及其潜在机制尚不清楚。
评价 YQFM 是否通过抑制 NMMHC IIA-肌动蛋白-ATG9A 相互作用形成自噬体，对脑缺血再灌注损伤发挥神经保护作用。
通过检测神经功能缺损、梗死体积和组织病理学变化，在大脑中动脉闭塞/再灌注（MCAO/R）模型小鼠（n=6）中研究 YQFM 的神经保护作用。采用免疫荧光共定位、免疫共沉淀和邻近连接分析检测 NMMHC IIA-肌动蛋白-ATG9A 相互作用。采用氧葡萄糖剥夺/复氧（OGD/R）模拟体外实验中的神经元，用于原代大鼠嗜铬细胞瘤（PC12）细胞。
在 MCAO/R 模型小鼠中，YQFM（1.342g/kg）通过调节 NMMHC IIA 介导的 ATG9A 转运，减轻脑缺血再灌注损伤。YQFM（400μg/ml）对 OGD/R 诱导的 PC12 细胞也具有类似作用。此外，NMMHC IIA 的 RNAi 削弱了 NMMHC IIA-F-肌动蛋白依赖性 ATG9A 转运，从而减弱了 YQFM 在体外的神经保护作用。
这些发现表明，YQFM 通过调节 NMMHC IIA-肌动蛋白-ATG9A 相互作用形成自噬体发挥神经保护作用。这一证据为 YQFM 在治疗脑缺血再灌注中的潜在机制提供了新的见解。
