Chen Li, Fan Xiao-Di, Qu Hua, Bai Rui-Na, Shi Da-Zhuo
Peking University Traditional Chinese Medicine Clinical Medical School (Xiyuan), Beijing 100191, China.
Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing 100191, China.
Evid Based Complement Alternat Med. 2021 Dec 31;2021:6518355. doi: 10.1155/2021/6518355. eCollection 2021.
Endothelial injury, characterized by an inflammatory response and increased permeability, is an initial stage of atherosclerosis (AS). Adenosine 5'-monophosphate (AMP), activated protein kinase (AMPK), and Nuclear Factor kappa B (NF-B)/Yin Yang 1(YY1) signaling pathways play important roles in the process of endothelial injury. Berberine (BBR), a bioactive alkaloid isolated from several herbal substances, possesses multiple pharmacological effects, including anti-inflammatory, antimicrobial, antidiabetic, anticancer, and antioxidant activities. Previous studies showed a protective effect of berberine against endothelial injury. However, the underlying mechanism remains unclear. We explored the potential effect of BBR on TNF- (tumor necrosis factor-) -induced injury of human umbilical endothelial cells (HUVECs) and studied its possible molecular mechanism. In the present study, HUVECs were divided into three groups. HUVEC viability was measured with Cell Counting Kit-8 assay. Extracellular lactic dehydrogenase (LDH) concentration was measured with LDH leakage assay. Endothelial microparticle (EMP) numbers were evaluated by flow cytometry analysis assay. The expression of proinflammatory cytokines was evaluated by Enzyme-Linked Immunosorbent Assay (ELISA). The mRNA expression of NF-B and YY1 was detected by Real-Time PCR (RT-PCR). The protein expression of NF-B, YY1, and AMPK was detected by immunofluorescence microscopy assay or western blot analysis. The results showed that LDH concentration, EMPs numbers, and the expression of proinflammatory cytokines (IL-6, IL-8, and IL-1) increased in TNF--induced injured HUVECs, but ameliorated by BBR pretreatment. BBR pretreatment upregulated the expression of phosphorylated AMPK and downregulated the expressions of NF-B and YY1 in injured HUVECs induced by TNF-, which were offset by the AMPK inhibitor Compound C (CC). The results indicated that BBR protected against TNF--induced endothelial injury via the AMPK/NF-B/YY1 signaling pathway.
以内皮炎症反应和通透性增加为特征的内皮损伤是动脉粥样硬化（AS）的初始阶段。5'-单磷酸腺苷（AMP）、活化蛋白激酶（AMPK）和核因子κB（NF-κB）/阴阳1（YY1）信号通路在内皮损伤过程中起重要作用。黄连素（BBR）是从多种草药中分离出的一种生物活性生物碱，具有多种药理作用，包括抗炎、抗菌、抗糖尿病、抗癌和抗氧化活性。先前的研究表明黄连素对内皮损伤具有保护作用。然而，其潜在机制仍不清楚。我们探讨了黄连素对肿瘤坏死因子-α（TNF-α）诱导的人脐静脉内皮细胞（HUVECs）损伤的潜在影响，并研究了其可能的分子机制。在本研究中，HUVECs被分为三组。采用细胞计数试剂盒-8法检测HUVECs活力。采用乳酸脱氢酶（LDH）泄漏法检测细胞外LDH浓度。通过流式细胞术分析评估内皮微粒（EMP）数量。采用酶联免疫吸附测定（ELISA）评估促炎细胞因子的表达。通过实时荧光定量聚合酶链反应（RT-PCR）检测NF-κB和YY1的mRNA表达。通过免疫荧光显微镜检测或蛋白质印迹分析检测NF-κB、YY1和AMPK的蛋白表达。结果显示，TNF-α诱导损伤的HUVECs中LDH浓度、EMPs数量以及促炎细胞因子（IL-6、IL-8和IL-1）的表达增加，但经黄连素预处理后有所改善。黄连素预处理上调了TNF-α诱导损伤的HUVECs中磷酸化AMPK的表达，下调了NF-κB和YY1的表达，而这被AMPK抑制剂化合物C（CC）所抵消。结果表明，黄连素通过AMPK/NF-κB/YY1信号通路保护细胞免受TNF-α诱导的内皮损伤。
