[Effect of acupuncture on intracerebral mitochondrial function in mice with cerebral hemorrhage].

Abstract

Objectives: To observe the effect of acupuncture on the recovery of neurological function in mice with intracerebral hemorrhage (ICH), so as to explore its mechanism of regulating mitochondria in brain tissue of ICH mice through the utilization of RNA sequencing and m6A methylation sequencing technology.

Methods: Twenty-four male C57BL/6J mice were ramondly divided into sham operation, model, and acupuncture groups (n=8 in each group). The ICH model was established by injection of the rat's auto-blood (10 μL) into the right caudatum region. Acupuncture group received manual acupuncture stimulation of "Baihui" (GV20) for 30 min, once every 12 h for 7 d. After the intervention, the behavioral scores of mice in each group were evaluated. The contents of glutathione peroxidase malondialdehyde (MDA), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) in brain tissue were detected by ELISA. The N6-methyladenosine (m6A) modification genes of ICH mice brain tissue were analyzed using RNA and m6A sequencing. The protein expression level of mitochondrial outer membrane translocase 20 (TOMM20) was detected by Western blot. The mRNA expression levels of Ndufb3、Ndufb6、Uqcr11、Cox7c、Ndufv1、Ndufb11 and Cytb were detected by PCR.

Results: Compared with the sham operation group, the suspension test score, balance walking test score, GSH-Px and SOD contents in the brain tissue were decreased (P<0.01), while the MDA content in the brain tissue was increased (P<0.01) in the model group. After the intervention and compared with the model group, the scores of the suspension test, the balance walking test and the contents of GSH-Px and SOD were increased (P<0.01, P<0.05), while the content of MDA was decreased (P<0.01) in the acupuncture group. RNA sequencing showed that a total of 780 differentially expressed genes were screened in the model group compared with the sham operation group, a total of 1 097 differentially expressed genes were screened in the acupuncture group compared with the model group, and the shared differential genes in the sham group vs model group and model group vs acupuncture group were 342;and 50 common differential genes were obtained after intersection with mitochondrial-related genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of the screened differentially expressed genes revealed that the above differential genes were mainly enriched in the oxidative phosphorylation pathway. PCR and Western blot verification results showed that compared with the sham operation group, the expression levels of Ndufb3, Ndufb6, Uqcr11, Cox7c, Ndufv1, Ndufb11 and Cytb mRNA, and the expression level of TOMM20 protein were decreased (P<0.01) in the model group. All the indexes mentioned above were reserved (P<0.01) in the acupuncture group.

Conclusions: Acupuncture can improve the neurological deficit signs of ICH mice, which maybe related to its function in regulating the mitochondrial function of brain tissue.