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. 2022 Apr 7:2022:2836880.
doi: 10.1155/2022/2836880. eCollection 2022.

Effects of Ruanmailing in Blocking Early Stages of Atherosclerosis by TNF- α Regulation via Kir2.1

Liufang Fan  1 , Xuanbin Huang  1 , Wenjuan Xue  1 , Weyan Xie  1 , Ruxi Tong  1 , Jinshui Chen  1 , Tianmin Wu  1
Affiliations

Effects of Ruanmailing in Blocking Early Stages of Atherosclerosis by TNF- α Regulation via Kir2.1

Liufang Fan et al. Evid Based Complement Alternat Med. .

Retraction in

Abstract

Objective: Ruanmailing oral solution consists of 16 herbs, has anti-lipid peroxidation activity, protects vascular endothelial cells, and improves vascular elasticity. It is an effective drug for the treatment of atherosclerosis (AS). The objective of this study was to investigate the mechanism underlying the antiatherosclerotic effects of Ruanmailing oral solution.

Methods: Macrophages were isolated, cultured, and divided into the macrophage control; macrophage foam cell; and low-, medium-, and high-concentration Ruanmailing groups. Cell proliferation was analyzed by cell counting kit-8 (CCK-8) assay, and the expression levels of inward-rectifier potassium ion channel 2.1 (Kir2.1) and tumor necrosis factor (TNF)-α were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analyses.

Results: CCK-8 assay results showed that the tested concentrations of Ruanmailing solution did not affect macrophage proliferation. RT-PCR and Western blot assays indicated that TNF-α expression increased significantly with the formation of macrophage foam cells (P < 0.05). In addition, significant decreases in Kir2.1 and TNF-α expression were observed following treatment with various concentrations of Ruanmailing (P < 0.05).

Conclusion: Based on the results, Ruanmailing affects macrophage foam cell formation by regulating Kir2.1 expression, which in turn reduces TNF-α expression and exerts antiatherosclerotic effects. These findings provide a scientific basis for the use of traditional Chinese medicine for AS treatment.

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Conflict of interest statement

The authors declare that they have no conflicts of interest.

Figures

Figure 1
Figure 1
Morphological changes in macrophages in each group. (a) Macrophage control group, (b) macrophage foam cell group, and (c) high-concentration Ruanmailing group. Magnification, ×100.
Figure 2
Figure 2
Trypan blue staining of macrophages in each group. (a) Macrophage control group, (b) macrophage foam cell group, and (c) high-concentration Ruanmailing group. Magnification, ×100.
Figure 3
Figure 3
Absorbance (optical density, OD) values of the different groups at different time points. Note. Comparison between groups at 0 h: aP > 0.05. Comparison of cell proliferation between macrophage control and macrophage foam cell groups at each time point, ∗P > 0.05. Comparison of cell proliferation among groups treated with different Ruanmailing concentrations at different time points, #P > 0.05. Comparison of cell proliferation between groups treated with different concentrations of Ruanmailing and the macrophage foam cell group at 24, 48, and 60 h: bP < 0.05. Group A: macrophage control, Group B: macrophage foam cell, Group C: low-concentration Ruanmailing, Group D: medium-concentration Ruanmailing, Group E: high-concentration Ruanmailing.
Figure 4
Figure 4
Detection of Kir2.1 and TNF-α protein expression. (a) TNF-α protein expression increased significantly after the formation of macrophage foam cells. (b) Kir2.1 and TNF-α protein expression levels decreased after treatment with serum containing different concentrations of Ruanmailing, and the differences were statistically significant. ImageJ was used for the quantitative analysis (∗P < 0.05). Group A: macrophage control, Group B: macrophage foam cell, Group C: low-concentration Ruanmailing, Group D: medium-concentration Ruanmailing, Group E: high-concentration Ruanmailing.
Figure 5
Figure 5
Gene expression levels of (a) TNF-α and (b) Kir2.1. TNF-α gene expression increased remarkably after the formation of macrophage foam cells. Treatment with serum containing different concentrations of Ruanmailing significantly decreased Kir2.1 and TNF-α gene expression levels compared with the levels in macrophage control (∗P < 0.05). Group A: macrophage control, Group B: macrophage foam cell, Group C: low-concentration Ruanmailing, Group D: medium-concentration Ruanmailing, Group E: high-concentration Ruanmailing.
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