YiQiFuMai lyophilized injection attenuates cerebral ischemic injury with inhibition of neuronal autophagy through intervention in the NMMHC IIA-actin-ATG9A interaction

Phytomedicine. 2022 Jan:95:153882. doi: 10.1016/j.phymed.2021.153882. Epub 2021 Dec 28.

Abstract

Background: 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.

Purpose: To evaluate whether YQFM has neuroprotective effects on cerebral ischemia/reperfusion-induced injury by inhibiting NMMHC IIA-actin-ATG9A interaction for autophagosome formation.

Methods: 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.

Results: 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.

Conclusion: 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.

Keywords: ATG9A; Autophagy; Cerebral ischemia/reperfusion; F-actin, Neuroprotective; Non-muscular myosin.

MeSH terms

  • Actins
  • Animals
  • Autophagy
  • Autophagy-Related Proteins
  • Brain Ischemia* / drug therapy
  • Drugs, Chinese Herbal* / pharmacology
  • Membrane Proteins
  • Mice
  • Neuroprotective Agents* / pharmacology
  • Rats
  • Reperfusion Injury* / drug therapy
  • Vesicular Transport Proteins

Substances

  • ATG9A protein, rat
  • Actins
  • Atg9A protein, mouse
  • Autophagy-Related Proteins
  • Drugs, Chinese Herbal
  • Membrane Proteins
  • Neuroprotective Agents
  • Vesicular Transport Proteins
  • yi-qi-fu-mai