Study on the mechanism of paeonol in improving low-density lipoprotein-induced human vascular endothelial cell injury

doi:10.12047/j.cjap.6325.2022.100

Abstract

Abstract: Objective: To investigate the effects of paeonol on low-density lipoprotein-induced human vascular endothelial cell injury and its molecular mechanisms. Methods: Human umbilical vein endothelial cells (HUVECs) were divided into 9 groups, normal control (NC) group, ox-LDL group (100 ng/L ox-LDL), low, medium, and high-dose paeonol groups (60 μmol/L, 120 μmol/L, 240 μmol/L paeonol+100 ng/L ox-LDL), ox-LDL+small interfering RNA negative control (si-NC) group, ox-LDL+circ_0003204 small interfering RNA (si-circ_0003204) group, middle dose group+ox-LDL+circ_0003204 overexpression negative control (pcDNA-NC) group, middle dose group+ox-LDL+circ_0003204 overexpression (pcDNA-circ_0003204) group, three replicate wells in each group. MTT flow cytometry, and Western blot were used to detect cell proliferation, apoptosis and protein (CDK2, Bcl2, p27, Bax) expressions, respectively. Malondialdehyde (MDA) and superoxide dismutase (SOD) kit were used to detect MDA content and SOD activity; real-time quantitative PCR (RT-qPCR) was used to detect the expression of circ_0003204. Results: Compared with the NC group, the proliferation activity, protein expressions of CDK2 and Bcl2, and SOD activity of HUVECs in the ox-LDL group were decreased significantly (P＜0.05), and the apoptosis rate, protein expressions of p27 and Bax, MDA content, and circ_0003204 expression were increased significantly (P＜ 0.05). Compared with the ox-LDL group, the proliferation activity, protein (CDK2, Bcl2) expressions and SOD activity of HUVECs in the low, medium and high dose paeonol groups were increased significantly (P＜0.05), and the apoptosis rate, protein (p27, Bax) expressions, MDA content And circ_0003204 expression were decreased significantly (P＜ 0.05). Compared with ox-LDL+si-NC group, the proliferation activity, protein (CDK2, Bcl2) expressions, SOD activity of HUVECs in ox-LDL+si-circ_0003204 group were increased significantly (P＜0.05), the apoptosis rate, protein (p27, Bax) expressions, and the content of MDA were decreased significantly (P＜0.05). Compared with the middle-dose+ox-LDL+pcDNA-NC group, the HUVECs proliferation activity, protein (CDK2, Bcl2) expressions, and SOD activity in the middle-dose+ox-LDL+pcDNA-circ_0003204 group were decreased significantly (P＜0.05), and the levels of circ_0003204, apoptosis rate, protein (p27, Bax) expressions and MDA content were increased significantly (P＜0.05). Conclusion: Paeonol can inhibit ox-LDL-induced apoptosis and oxidative stress of human umbilical vein endothelial cells, and alleviate human umbilical vein endothelial cell injury. The mechanism of action may be related to the down-regulation of circ_0003204 expression.

Key words: paeonol, circular RNA 0003204, atherosclerosis, human umbilical vein endothelial cells, proliferation, apoptosis, oxidative stress

CLC Number:

R285

JIA Cheng-wen, LI Ying-dong, JIANG Hu-gang, HAN Guo-wei, ZHAO Xin-ke. Study on the mechanism of paeonol in improving low-density lipoprotein-induced human vascular endothelial cell injury[J]. CJAP, 2022, 38(5): 537-542.

References

[1] 高鹏琳, 桂丽卿, 袁奕珂, 等. 中医药治疗动脉粥样硬化研究进展[J]. 神经病学与神经康复学杂志, 2018, 14(4): 55-60.
[2] 吴洁, 宋耀鸿. 中医药保护动脉粥样硬化血管内皮细胞研究进展[J]. 中医学, 2020, 9(2): 115-121.
[3] 胡云飞, 徐国兵. 牡丹皮及其主要成分丹皮酚的药理作用研究进展[J]. 安徽医药, 2014, 18(4): 589-592.
[4] 蒋丽丽, 张彦龙, 王春杰, 等. 牡丹皮中有效成分丹皮酚的药理活性研究进展[J]. 现代诊断与治疗, 2016, 27(22): 4223-4224.
[5] 龚明玉, 许倩, 李素婷. 丹皮酚对动脉粥样硬化大鼠的防治作用及其机制[J]. 中成药, 2018, 40(2): 437-440.
[6] 孙颖, 刘雅蓉, 吴鸿飞, 等. 丹皮酚对动脉粥样硬化相关细胞的作用及机制[J]. 安徽中医学院学报, 2020, 39(3): 92-96.
[7] 孙艺丹, 侯晓峰. 丹皮酚对H2O2诱导血管内皮细胞损伤的保护作用[J]. 中国处方药, 2019, 17(4): 37-38.
[8] Zhang DY, Zhang G, Yu K, et al. Circ_0003204 knockdown protects endothelial cells against oxidized low-density lipoprotein-induced injuries by targeting the miR-491-5p-ICAM1 pathway[J]. J Thromb Thrombolys, 2022, 53(2): 302-312.
[9] Liu H, Ma X, Mao Z, et al. Circular RNA has_circ_0003204 inhibits ox-LDL-induced vascular endothelial cell proliferation and angiogenesis[J]. Cell Signal, 2020, 70(1): 109595-109602.
[10] 刘雅蓉, 陈君君, 戴敏, 等. 丹皮酚通过miR-21介导的p38MAPK信号通路下调氧化低密度脂蛋白诱导的大鼠血管内皮细胞肿瘤坏死因子-α释放[J]. 安徽中医学院学报, 2014, 33(1): 51-56.
[11] 高爽, 王臻楠, 顾耘. 血管内皮细胞功能障碍与动脉粥样硬化关系的研究进展[J]. 中西医结合心脑血管病杂志, 2018, 16(20): 50-54.
[12] 陈刚, 陈九霖, 吴俊. miR-133b靶向抑制SGTB对oxLDL诱导的血管内皮细胞损伤的影响[J]. 中国应用生理学杂志, 2021, 37(6): 594-600.
[13] 欧阳昕, 李清, 肖爱娇, 等. 艾灸对缺氧缺血性脑损伤新生小鼠的治疗作用[J]. 中国应用生理学杂志, 2021, 37(5): 543-547.
[14] Bassi C, Fortin J, Snow BE, et al. The PTEN and ATM axis controls the G1/S cell cycle checkpoint and tumorigenesis in HER2-positive breast cancer[J]. Cell Death Differ, 2021, 28(11): 3036-3051.
[15] 李杰. 中医药治疗动脉粥样硬化研究进展[J]. 内蒙古中医药, 2019, 38(12): 167-168.
[16] 周晓慧, 牛成伟, 曹凯, 等. 丹皮酚通过抑制NF-κB信号通路下调高脂血清诱导的人脐静脉内皮细胞黏附分子的表达[J]. 中国病理生理杂志, 2011, 27(2): 249-253.
[17] 胡文君, 张振, 戴敏. 丹皮酚通过抑制PI3K/AKT-NF-κB通路对LPS诱导的与平滑肌细胞共培养的大鼠血管内皮细胞的保护作用[J]. 中国中药杂志, 2016, 41(12): 2298-2302.
[18] Qin M, Wang W, Zhou H, et al. Circular RNA circ_0003645 silencing alleviates inflammation and apoptosis via the NF-κB pathway in endothelial cells induced by oxLDL[J]. Gene, 2020, 755(1): 144900-144907.
[19] Zhang W, Sui Y.CircBPTF knockdown ameliorates high glucose-induced inflammatory injuries and oxidative stress by targeting the miR-384/LIN28B axis in human umbilical vein endothelial cells[J]. Mol Cell Biochem, 2020, 471(1-2): 101-111.