Role of NADPH oxidase in total salvianolic acid injection attenuating ischemia-reperfusion impaired cerebral microcirculation and neurons: implication of AMPK/Akt/PKC.
OBJECTIVE
TSI is a new drug derived from Chinese medicine for treatment of ischemic stroke in China. The aim of this study was to verify the therapeutic effect of TSI in a rat model of MCAO, and further explore the mechanism for its effect.


METHODS
Male Sprague-Dawley rats were subjected to right MCAO for 60 minutes followed by reperfusion. TSI (1.67 mg/kg) was administrated before reperfusion via femoral vein injection. Twenty-four hours after reperfusion, the fluorescence intensity of DHR 123 in, leukocyte adhesion to and albumin leakage from the cerebral venules were observed. Neurological scores, TTC staining, brain water content, Nissl staining, TUNEL staining, and MDA content were assessed. Bcl-2/Bax, cleaved caspase-3, NADPH oxidase subunits p47(phox)/p67(phox)/gp91(phox), and AMPK/Akt/PKC were analyzed by Western blot.


RESULTS
TSI attenuated I/R-induced microcirculatory disturbance and neuron damage, activated AMPK, inhibited NADPH oxidase subunits membrane translocation, suppressed Akt phosphorylation, and PKC translocation.


CONCLUSIONS
TSI attenuates I/R-induced brain injury in rats, supporting its clinic use for treatment of acute ischemic stroke. The role of TSI may benefit from its antioxidant activity, which is most likely implemented via inactivation of NADPH oxidase through a signaling pathway implicating AMPK/Akt/PKC.
The Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, China.
Hao
H
Tang
Chun-Shui
CS
Pan
Xiao-Wei
XW
Mao
Yu-Ying
YY
Liu
Li
L
Yan
Chang-Man
CM
Zhou
Jing-Yu
JY
Fan
Shuang-Yan
SY
Zhang
Jing-Yan
JY
Han
D055372:AMP-Activated Protein Kinases / Q000502:physiology; D000475:Alkenes / Q000494:pharmacology / Q000627:therapeutic use*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002199:Capillary Permeability / Q000187:drug effects; D002544:Cerebral Infarction / Q000209:etiology / Q000473:pathology / Q000517:prevention & control; D004365:Drugs, Chinese Herbal / Q000494:pharmacology / Q000627:therapeutic use*; D020244:Infarction, Middle Cerebral Artery / Q000188:drug therapy* / Q000201:enzymology / Q000503:physiopathology; D007962:Leukocytes / Q000187:drug effects; D015227:Lipid Peroxidation / Q000187:drug effects; D008297:Male; D008833:Microcirculation / Q000187:drug effects*; D009069:Movement Disorders / Q000209:etiology / Q000517:prevention & control; D019255:NADPH Oxidases / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000502:physiology; D009474:Neurons / Q000187:drug effects* / Q000201:enzymology; D010766:Phosphorylation / Q000187:drug effects; D059808:Polyphenols / Q000494:pharmacology / Q000627:therapeutic use*; D011493:Protein Kinase C / Q000502:physiology; D021381:Protein Transport / Q000187:drug effects; D051057:Proto-Oncogene Proteins c-akt / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D015427:Reperfusion Injury / Q000188:drug therapy* / Q000201:enzymology / Q000503:physiopathology; D015398:Signal Transduction / Q000187:drug effects
D000475:Alkenes; D004365:Drugs, Chinese Herbal; D009419:Nerve Tissue Proteins; D059808:Polyphenols; D017382:Reactive Oxygen Species; C568740:salvianolic acid; D019255:NADPH Oxidases; C494920:Akt1 protein, rat; D051057:Proto-Oncogene Proteins c-akt; D011493:Protein Kinase C; D055372:AMP-Activated Protein Kinases
AMPK/Akt/PKC; NADPH; antiapoptosis; antioxidation; microvasculature