鹿角方调控NF-κB通路抑制血管紧张素 Ⅱ诱导的心肌细胞肥大

doi:10.19803/j.1672-8629.20240494

摘要/Abstract

摘要： 目的探究鹿角方调控NF-κB信号通路对血管紧张素Ⅱ（Ang Ⅱ）诱导的心肌细胞肥大的影响。方法以血管紧张素Ⅱ诱导建立原代乳鼠心肌细胞肥大模型,将细胞分为正常组、Ang Ⅱ模型组、鹿角方+Ang Ⅱ组、BAY11-7082+Ang Ⅱ组、鹿角方+BAY11-7082+Ang Ⅱ组。检测终浓度为0、0.1、0.3、1、3、10μg·mL^-1的鹿角方对心肌细胞表面积的影响;酶联免疫吸附法（ELISA）检测有效浓度10、100μg·mL^-1鹿角方对心肌细胞培养液中肿瘤坏死因子α（TNF-α）、白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和白细胞介素-2（IL-2）的含量的影响;通过实时荧光定量聚合酶链反应（qRT-PCR）检测ANP和BNP的基因表达;蛋白免疫印迹（Western Blot）法检测NF-κB p65、p-NF-κB p65（Ser536）、p-IKK（Ser176/180）、IκBα和p-IκBα（Ser32）的蛋白表达。结果与正常组相比,模型组心肌细胞表面积显著增大（P<0.01或P<0.05）,肥大标志物ANP和BNP mRNA表达显著增加（P<0.01）,促炎因子TNF-α、IL-1β、IL-6的水平显著升高（P<0.01）,抑炎因子IL-2的水平显著降低（P<0.01）,p-NF-κB p65（Ser536）、p-IKK（Ser176/180）和p-IκBα（Ser32）蛋白表达均显著增加（P<0.01）,IκBα蛋白表达显著下降（P<0.01）;与模型组相比,鹿角方可显著抑制心肌细胞表面积增大（P<0.01）,显著抑制心肌细胞ANP和BNP mRNA表达（P<0.01）,显著降低心肌细胞培养液中TNF-α、IL-1β和IL-6水平（P<0.01）,显著升高IL-2的水平（P<0.01）,并显著降低p-NF-κB p65（Ser536）、p-IKK（Ser176/180）和p-IKBα（Ser32）的蛋白表达（P<0.01或P<0.05）,显著增加IκBα蛋白表达水平（P<0.01）,且鹿角方对上述蛋白表达水平的影响与BAY11-7082相当（P>0.05）。结论鹿角方可显著抑制Ang Ⅱ诱导的心肌细胞肥大,其机制与阻断NF-κB信号通路有关。

关键词: 鹿角方, 心肌细胞, 血管紧张素Ⅱ, 心力衰竭, 心肌细胞肥大, NF-κB信号通路, 炎症, SD 乳鼠

Abstract: Objective To investigate the effect of Lujiao formula on cardiac hypertrophy induced by angiotensin II (Ang II) through regulating the NF-κB signaling pathway. Methods Ang II was used to establish a cardiac hypertrophy model of neonatal rat cardiac myocytes (NRCMs). The cells were divided into normal group, Ang Ⅱ model group, Lujiao formula + Ang Ⅱ group, BAY11-7082 + Ang Ⅱ group and Lujiao formula + BAY11-7082 + Ang Ⅱ group. The effect of Lujiao formula with final concentrations of 0, 0.1, 0.3, 1, 3, and 10 μ g · mL^-1 on the surface area of cardiomyocytes was detected. The level of tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6) and interleukin-2 (IL-2) in culture medium of cells treated with Lujiao formula at effective concentrations of 10 and 100 μ g · mL^-1 were detected by enzyme-linked immunosorbent assay (ELISA). Quantitative real-time polymerase chain reaction (RT-qPCR) was used to quantify the gene expression of ANP and BNP. Protein expression of NF-κB p65, p-NF-κB p65 (Ser536), p-IKK (Ser176/180), IκBα and p-IκBα (Ser32) were detected by Western Blot. Results Compared with normal group, the surface area of cardiomyocytes in model group was significantly increased (P<0.01 or P<0.05), and the mRNA level of hypertrophy markers ANP and BNP were increased (P<0.01), and the levels of pro-inflammatory factors TNF-α, IL-1β and IL-6 were increased (P<0.01). The level of anti-inflammatory factor IL-2 was decreased (P<0.01), the protein expressions of p-NF-κBp65 (Ser536), p-IKK (Ser176/180) and p-IκBα (Ser32) were significantly increased (P<0.01), and the protein expressions of IκBα were decreased (P<0.01). Compared with the model group, Lujiao formula could significantly inhibit the increase of surface area of cardiomyocytes (P<0.01), and inhibit the expressions of ANP and BNP mRNA of cardiomyocytes (P<0.01), and reduce the levels of TNF-α, IL-1β and IL-6 in cardiomyocyte culture medium (P<0.01), and increase the level of IL-2 (P<0.01). Lujiao formula could significantly decrease the protein expressions of p-NF-κB p65 (Ser536), p-IKK (Ser176/180) and p-IκBα(Ser32), and increase the protein expressions of IκBα (P<0.01 or P<0.05), and the effect of Lujiao formula on these protein levels was comparable to that of BAY11-7082 (P>0.05). Conclusion Lujiao formula could significantly inhibit cardiac hypertrophy induced by angiotensin II through inhibiting the NF-κB signaling pathway.

Key words: Lujiao Formula, Cardiomyocytes, Ang II, Heart Failure, Cardiac Hypertrophy, NF-κB Signaling Pathway, Inflammation, SD Neonatal Rat

中图分类号:

张建伟, 刘力, 徐光临, 时潇丽. 鹿角方调控NF-κB通路抑制血管紧张素 Ⅱ诱导的心肌细胞肥大[J]. 中国药物警戒, 2024, 21(11): 1250-1255.

ZHANG Jianwei, LIU Li, XU Guanglin, SHI Xiaoli. Lujiao Formula Inhibiting Cardiac Hypertrophy Induced by Ang Ⅱ through Regulating the NF-κB Pathways[J]. Chinese Journal of Pharmacovigilance, 2024, 21(11): 1250-1255.

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