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

JOURNAL OF BEIJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE ›› 2020, Vol. 43 ›› Issue (8): 653-660.doi: 10.3969/j.issn.1006-2157.2020.08.007

• Chinese Medicinal Pharmacology • Previous Articles     Next Articles

Regulation of input protein α3/NF-κB pathway by adjusting miR-181 with dissipating phlegm and removing blood stasis method and its anti-atherosclerosis mechanism*

Qin Hewei1,2, Li Yanjie1, Zhang Zhixin1#, Ji Lingshan1, Guo Ning2   

  1. 1 Henan Province Hospital of & TCM Second Affiliated Hospital of Henan University of Chinese Medicine, Henan 450002, China;
    2 Henan University of Chinese Medicine, Henan 450002, China
  • Received:2020-01-29 Online:2020-08-30 Published:2020-08-27
  • Contact: Zhang Zhixin, M.D., Deputy Chief Physician. No. 6 Dongfeng Road, Zhengzhou, Henan Province Hospital of TCM, Henan 450002. E-mail: qinheweii@126.com
  • Supported by:
    National Natural Science Foundation of China (No.81704030)

Abstract: Objective To observe the correlation between the anti-atherosclerosis mechanism of dissipating phlegm and removing blood stasis method and the effect on the input protein α3/NF-κB signaling pathway through the regulation of microribonucleic acid 181 (miR-181). Methods In vitro experiments, vascular endothelial cell injury model was established with oxidized low-density lipoprotein (ox-LDL). According to different intervention methods, cells were randomly divided into four groups: model group (blank serum), miR-181 mimic group, miR -181 inhibitor group, phlegm-resolving and blood stasis-eliminating TCM serum group (TCM serum group). After intervention, the cell activity of each group was detected. RT-PCR and Western blot were used to detect the mRNA and protein expression levels of miR-181, input protein α3 and NF-κB. In vivo experiments, 60 ApoE-/- mice were randomly divided into 5 groups: model group, miR-181 inhibitor group, miR-181 mimic group, high-dose and low-dose phlegm-resolving and blood stasis-eliminating TCM serum group (TCM high-dose serum group, and TCM low-dose serum group respectively) (n=12). After continuous intervention for 8 weeks, the plasma and aorta were collected for testing. The levels of IL-6, VCAM-1 and E-selectin in the peripheral plasma were tested by ELISA. The expression of microRNA-181, input protein α3 and NF-κB in the aortic wall of each group was tested by RT-PCR, and the pathological changes were observed. Results In vitro experiments, the cell activity of the TCM serum group and the miR-181 mimic group was significantly increased (P<0.05), and the expression of α3 and NF-κB mRNA were significantly down-regulated (P<0.05). In vivo experiments, compared with the model group, the plasma levels of IL-6, VCAM-1 and E-selectin in the TCM high-dose and low-dose serum groups and miR-181 mimic group were significantly lower, compared with the model group (P<0.05); compared with the inhibitor group, the levels of peripheral plasma IL-6, VCAM-1 and E-selectin in the TCM serum group were lower (P<0.05). Compared with the model group, the mRNA expression of aortic miR-181 in the TCM high-dose and low-dose serum groups and miR-181 mimic group was significantly increased (P<0.05), and the expression level of α3 and NF-κB significantly down-regulated (P<0.05). There was no significant difference in the indexes between the TCM high-dose and low-dose serum groups (P>0.05). Conclusion The anti-atherosclerosis mechanism of resolving phlegm and eliminating blood stasis may be related to the targeted regulation of miR-181 to affect the input protein α3/NF-κB signaling pathway, which could possibly reduce the levels of inflammatory mediators such as IL-6, VCAM-1 and E-selectin, thus reducing the damage of vascular endothelial cells.

Key words: atherosclerosis, phlegm-resolving and blood stasis-eliminating method, endothelial cells, miR-181, input protein α3, NF-κB, mice

CLC Number: 

  • R285.5