主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

JOURNAL OF BEIJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE ›› 2021, Vol. 44 ›› Issue (4): 366-373.doi: 10.3969/j.issn.1006-2157.2021.04.012

• Acupuncture & Moxibustion • Previous Articles     Next Articles

Protective mechanism of electroacupuncture serum on neurons after cerebral ischemia based on H4K16AC-mediated autophagy*

Xu Shuying1, Shen Yan2, Peng Yongjun1#, Yang Sha2, Li Wenqian1   

  1. 1 Hospital affiliated to Nanjing University of Chinese Medicine, Jiangsu 210029, China;
    2 First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, China National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion,Tianjin 300192, China
  • Received:2020-05-18 Published:2021-04-29
  • Supported by:
    National Natural Science Foundation of China (No. 81973932, No.81574060, No.81001556); the Tianjin Natural Science Foundation (18JCZDJC99200); Developmental Program for Changjiang Scholars and Innovative Research Team Program (IRT1167)

Abstract: Objective To study the protective mechanism of electroacupuncture serum on neurons after cerebral ischemia from the perspective of autophagy regulation by acetylation of histone H4 16 lysine (H4K16ac). Methods 1) Preparation of electroacupuncture serum: The improved rat model of acute focal cerebral ischemia reperfusion was established. Electroacupuncture intervention was performed 5 at min and 16 h after the model was successfully established. Acupuncture was performed at the points of Renzhong (GV 26) and Baihui (GV 20), and electroacupuncture treatment was applied for 30 mins. The samples were collected 7 h after the second electroacupuncture treatment, and the serum of the rats was used for reserve. 2) Cell experiment: The rat cerebral cortex neuron cells were divided into control group, model group, electroacupuncture group, hMOF inhibitor group and Sirt1 inhibitor group. All cells were deprived of oxygen and sugar for 3 h and then re-oxygenated and re-glycated for 24 h except the control group. Meanwhile, all these cells were treated with 2% electroacupuncture serum, 15 g/L hMOF siRNA and 8 g/L Sirt1 siRNA. Cell proliferation was measured by using CCK-8 after 48 h of cell treatment. Cell apoptosis was measured by using flow cytometry after 48 h treatment. hMOF, Sirt1, H4K16ac, LC3-Ⅱ, and Beclin1 protein expression in rat cerebral cortex neurons were measured with Western blot assay. The expression of hMOF, Sirt1 and Beclin1 mRNA was detected by using qPCR. ChIP was used to detect the binding degree of H4K16ac of neurons in each group in the Beclin1 promoter region of autophagy target gene. Results Compared with the model group, the cell proliferation activity was significantly increased (P<0.05), and the cell apoptosis rate was significantly decreased (P<0.05). Compared with model group, both electroacupuncture and hMOF inhibitor decreased the expression of hMOF and H4K16ac proteins (P<0.05), and increased the expression of Sirt1, LC3-II and Beclin1 (P<0.05). There was no significant difference between Sirt1 inhibitor group and model group (P>0.05). According to qPCR results, compared with model group, both electroacupuncture and hMOF inhibitor could inhibit hMOF mRNA expression (P<0.05), while Sirt1 and Beclin1 mRNA expression were up-regulated (P<0.05). Compared with the model group, the enrichment of H4K16ac in the Beclin1 promoter region of autophagy target gene was significantly increased in the electroacupuncture group (P<0.05). Conclusion Electroacupuncture serum has a protective effect on OGD/R injured neurons, and its anti-cerebral ischemia reperfusion injury may be that electroacupuncture regulates autophagy by regulating the expression of hitin H4K16ac, thereby reducing cerebral ischemia reperfusion injury and playing a protective effect on nerve cells.

Key words: electroacupuncture serum, rat cerebral cortex neuron, H4K16ac, hMOF, histone deactylase inhibitor Sirt1, cell autophagy

CLC Number: 

  • R245