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Journal of Peking University (Health Sciences) ›› 2022, Vol. 54 ›› Issue (4): 599-604. doi: 10.19723/j.issn.1671-167X.2022.04.004

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Effects of 4′-O-methylochnaflavone on endothelial dysfunction induced by palmitic acid in rat cavernous endothelial cells

Yang-yang GU1,Xiao-hui TAN1,Wen-peng SONG1,Dong FANG1,Wei-dong SONG1,Yi-ming YUAN1,Ning-han FENG2,*(),Rui-li GUAN1,*()   

  1. 1. Department of Urology, Peking University First Hospital; Institute of Urology, Peking University; Beijing 100034, China
    2. Department of Urology, Wuxi Second Hospital, Nanjing Medical University, Wuxi 214002, Jiangsu, China
  • Received:2022-03-25 Online:2022-08-18 Published:2022-08-11
  • Contact: Ning-han FENG,Rui-li GUAN E-mail:n.feng@njmu.edu.cn;guanruili@bjmu.edu.cn
  • Supported by:
    the National Natural Science Foundation of China(81971379);the Wuxi "Taihu Talents Program" Medical and Health High-Level Talents Project

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Abstract:

Objective: To investigate the effect of biflavonoid 4′-O-methylochnaflavone (MF) on palmitic acid-induced endothelial dysfunction in rat cavernous endothelial cells (RCECs). Methods: The isolated RCECs were commercially available and randomly divided into four groups: normal+BSA group (NC group), palmitic acid (PA) group, MF group, and icariside Ⅱ (ICA Ⅱ) group. The protein expression levels of protein kinase B (PKB/AKT) and endothelial nitric oxide synthase (eNOS) in each group were evaluated via Western blotting. The differences in the intracellular nitric oxide of RCECs treated by MF or ICA Ⅱ were detected by DAF-FM DA that served as a nitric oxide fluorescent probe. Effects of MF and ICA Ⅱ on cell proliferation of PA-stimulated RCECs were determined via CCK-8 assay. Results: The content of nitric oxide in RCECs was significantly increased after the treatment of MF and ICA Ⅱ in comparison with the NC group (P < 0.05). Moreover, compared with ICA Ⅱ group, MF demonstrated a more obvious effect in promoting nitric oxide production (P < 0.05). Compared with the NC group, the expression levels of eNOS and AKT in the PA group were significantly decreased, indicating that a model for simulating the high-fat environment in vitro was successfully constructed (P < 0.05). Meanwhile, the intervention of MF and ICA Ⅱ could effectively increase the expression of eNOS and AKT, suggesting that MF and ICA Ⅱ could promote the recovery of endothelial dysfunction caused by high levels of free fatty acids (P < 0.05). The results of CCK-8 assays showed that PA could significantly reduce the proli-feration ability of RCECs (P < 0.05). Furthermore, the decreased cell viability induced by PA was significantly elevated by treatment with ICA Ⅱ and MF (P < 0.05). Conclusion: In RCECs, MF and ICA Ⅱ could effectively increase the content of nitric oxide. The down-regulation of the expression of proteins associated with the AKT/eNOS pathway after PA treatment revealed that this pathway was involved in the development of endothelial dysfunction, which could be effectively reversed by MF and ICA Ⅱ. In addition, the cell proliferation ability was significantly decreased following PA treatment, but MF and ICA Ⅱ could restore the above changes. Overall, biflavonoid MF has an obvious repairing effect on PA-stimulated endothelial dysfunction.

Key words: Endothelial dysfunction, Palmitic acid, Nitric oxide synthase, Signal transduction

CLC Number: 

  • R698.1

Figure 1

Chemical structure of 4′-O-methylochnaflavone and icariside Ⅱ"

Figure 2

The evaluation of intracellular NO following the treatment of MF and ICA Ⅱ"

Figure 3

The protein level of eNOS and AKT following the treatment of PA PA, palmitic acid; AKT, protein kinase B; eNOS, endothelial nitric oxide synthase. *P < 0.05."

Figure 4

The protein level of eNOS and AKT in the PA-stimulated cells following the treatment of MF and ICA Ⅱ NC, normal control; PA, palmitic acid; MF, 4′-O-methylochnaflavone; ICA Ⅱ, icariside Ⅱ; AKT, protein kinase B; eNOS, endothelial nitric oxide synthase. *P < 0.05."

Figure 5

The RCECs proliferation ability of the PA-stimulated cells following the treatment of MF and ICA Ⅱ A, the RCECs were treated with different concentrations of MF and ICA Ⅱ for 24 h with cell viability measured via the CCK-8 assay; B, the RCECs were treated with 150 μmol/L PA in the presence or absence of MF and ICA Ⅱ for 24 h with cell viability measured via the CCK-8 assay. CCK, cell counting kit; NC, normal control; PA, palmitic acid; MF, 4′-O-methylochnaflavone; ICA Ⅱ, icariside Ⅱ; RCECs, rat cavernous endothelial cells; NS, no significant; μM, μmol/L; ***P < 0.001."

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