Effect of ethanol extract of Gastrodiae Rhizoma on mitochondrial dysfunction in cerebral ischemia-reperfusion injury.
10.19540/j.cnki.cjcmm.20220613.708
- Author:
Yuan LUO
1
;
Pu CHEN
1
;
Li-Ping YANG
1
;
Xiao-Hua DUAN
1
Author Information
1. Yunnan University of Traditional Chinese Medicine,Yunnan Key Laboratory of Dai Medicine and Yi Medicine Kunming 650500, China.
- Publication Type:Journal Article
- Keywords:
Gastrodiae Rhizoma ethanol extract;
apoptosis;
cerebral ischemia-reperfusion injury;
mitochondrial dysfunction;
oxidative stress
- MeSH:
Animals;
Rats;
bcl-2-Associated X Protein/metabolism*;
Reactive Oxygen Species/metabolism*;
Ethanol;
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine/pharmacology*;
Neuroprotective Agents/pharmacology*;
Rats, Sprague-Dawley;
Reperfusion Injury/metabolism*;
Infarction, Middle Cerebral Artery;
Mitochondria;
Proto-Oncogene Proteins c-bcl-2/metabolism*;
Apoptosis;
Cytochromes c/metabolism*;
Adenosine Triphosphate/pharmacology*
- From:
China Journal of Chinese Materia Medica
2022;47(20):5564-5573
- CountryChina
- Language:Chinese
-
Abstract:
This study observed the effects of ethanol extract of Gastrodiae Rhizoma(GE) on multiple aspects of mitochondrial dysfunction by investigating the mitochondria isolated from rat brains subjected to focal middle cerebral artery occlusion/reperfusion(MCAO/R). SD rats were randomly divided into a sham operation group(Sham), a model group(MCAO/R), low-and high-dose ethanol extract of GE groups(262.3 and 524.6 mg·kg~(-1)), and a nimodipine group(Nim, 15 mg·kg~(-1)). After continuous intragastric administration for 7 days, the MCAO/R model was induced in rats by the suture method. The neurological function and percentage of cerebral infarction volume were measured 24 h after reperfusion, and mitochondrial ultrastructure was observed under an electron microscope. Mitochondria were separated by gradient centrifugation and detected for reactive oxygen species(ROS), malondialdehyde(MDA), respiratory chain enzyme complex Ⅰ-Ⅳ activity, mitochondrial permeability transition pore(mPTP), mitochondrial membrane potential(MMP), and mitochondrial adenosine triphosphate(ATP) content. Enzyme-linked immunosorbent assay(ELISA) was used to detect the expression of cytochrome C(Cyt C) in different sites. TUNEL staining was used to observe the apoptosis of brain tissues in each group, and Western blot was used to detect the expression of B-cell lymphoma 2(Bcl-2) and Bcl-2-associated X protein(Bax) in brain tissues. The experimental results revealed that compared with the Sham group, the MCAO/R group showed evident neurological dysfunction and cerebral infarction(P<0.01) accompanied by mitochondrial swelling and crest disappearance, increased ROS level and MDA content, inhibited activity of respiratory chain enzyme complex Ⅰ-Ⅳ, abnormal opening of mPTP, and reduced MMP and mitochondrial ATP(P<0.01). Besides, many Cyt C was released from mitochondria into the cytoplasm to induce apoptosis(P<0.01). The ethanol extract of GE positively affected the behavior deficit and mitochondrial health of MCAO/R rats, with the manifestations of decreased production of ROS and MDA(P<0.01), potentiated activity of mitochondrial respiratory chain enzyme complex Ⅰ-Ⅳ, and restored the level of mPTP(P<0.05). In addition, the ethanol extract of GE reduced the loss of MMP and the escape of Cyt C to the cytoplasm(P<0.05), reduced the number of TUNEL positive cells(P<0.01) accompanied by the decrease in Bax and the up-regulation of Bcl-2(P<0.01), and increased the level of ATP(P<0.01). In conclusion, GE ethanol extract has a protective effect against MCAO/R-induced mitochondrial dysfunction, and the mechanism may be related to the regulation of oxidative stress, maintenance of functional morphology of mitochondria, and inhibition of endogenous apoptosis.
