Fan Qinghong, Liu Yujuan, Rao Jiaoyu, Zhang Zhe, Xiao Wei, Zhu Tao, Chai Xiaomeng, Ye Kaihe, Ning Na, Yin Zhen, Chai Yushuang, Xu Yimin, Lan Ruirui, Verkhratsky A, Nie Hong
Guangdong Province Key Laboratory of Pharmacodynamic Constituents of Traditional Chinese Medicine and New Drugs Research, College of Pharmacy, Jinan University, Guangzhou, China.
Guangzhou Baiyunshan Zhongyi Pharmaceutical Co., Ltd, Guangzhou, China.
Front Pharmacol. 2020 Jan 31;10:1584. doi: 10.3389/fphar.2019.01584. eCollection 2019.
Angong Niuhuang Pill (ANP) is a well-known patented Chinese medicine which is used for hundreds of years for treating the central nervous system diseases. Atherosclerosis is a poly-aetiological chronic inflammatory vascular disease. Preventing inflammation is fundamental for treating atherosclerosis in early stages. In this study, we investigated the protective effects and possible mechanisms of ANP action on a high-fat diet induced early and mid-term atherosclerosis ApoE mice. The effects of ANP were compared with accepted drug simvastatin. Twelve male C57BL/6J mice were used as the control group, and 60 male ApoE mice were randomly divided into five groups: Model group, Simvastatin group, Low-, Medium-, and High-dose ANP group these groups received, respectively, saline, simvastatin (3.0mg/kg), low-dose ANP (0.25 g/kg), medium-dose ANP (0.50 g/kg), and high-dose ANP (1.0 g/kg), once every other day for 10 weeks. After administration, serum biochemical indices were detected by the automatic biochemical analyzer, the concentrations of IL-6 and IL-10 in the serum were assayed by ELISA, expression levels of IL-1β, TNF-α, MMP-2, MMP-9, CCL2, and its receptor CCR2 in the full-length aorta, and expression levels of transcription factors Foxp3, RORγt in the spleen were assayed western blotting and RT-qPCR. Flow cytometry was used to analyze Th17 cells and Treg cells. Pathological and histological analysis was completed on aortic root. ANP decreased LDL/HDL ratio, concentrations of IL-6 while increased IL-10 in serum. Moreover, ANP down-regulated the expression levels of IL-1β, TNF-α, MMP-2, MMP-9, CCL2, and CCR2 receptor in the full-length aorta. In addition, ANP decreased Th17 cells and expression levels of transcription factor RORγt, increased Treg cells and expression levels of transcription factor Foxp3. ANP decreased content of collagen fibers and infiltration of inflammatory cells in the aortic root. In conclusion, we demonstrated that ANP has anti-atherosclerosis effects on a high-fat diet induced ApoE mice early and mid-term AS model regulating Th17/Treg balance, inhibiting chronic inflammation, reducing plaque collagen fibers, and reducing inflammatory cells infiltration, to exert its multi-channel multi-target anti-early and mid-term AS effects.
安宫牛黄丸（ANP）是一种著名的专利中药，数百年来一直用于治疗中枢神经系统疾病。动脉粥样硬化是一种多病因的慢性炎症性血管疾病。预防炎症是早期治疗动脉粥样硬化的根本。在本研究中，我们研究了安宫牛黄丸对高脂饮食诱导的早期和中期动脉粥样硬化ApoE小鼠的保护作用及可能机制。将安宫牛黄丸的作用与公认药物辛伐他汀进行比较。12只雄性C57BL/6J小鼠作为对照组，60只雄性ApoE小鼠随机分为五组：模型组、辛伐他汀组、低、中、高剂量安宫牛黄丸组，这些组分别接受生理盐水、辛伐他汀（3.0mg/kg）、低剂量安宫牛黄丸（0.25g/kg）、中剂量安宫牛黄丸（0.50g/kg）和高剂量安宫牛黄丸（1.0g/kg），每隔一天给药一次，共10周。给药后，用自动生化分析仪检测血清生化指标，用ELISA法检测血清中IL-6和IL-10的浓度，用Western blotting和RT-qPCR法检测全长主动脉中IL-1β、TNF-α、MMP-2、MMP-9、CCL2及其受体CCR2的表达水平，以及脾脏中转录因子Foxp3、RORγt的表达水平。用流式细胞术分析Th17细胞和Treg细胞。对主动脉根部进行病理和组织学分析。安宫牛黄丸降低了血清中LDL/HDL比值、IL-6浓度，同时升高了IL-10浓度。此外，安宫牛黄丸下调了全长主动脉中IL-1β、TNF-α、MMP-2、MMP-9、CCL2和CCR2受体的表达水平。此外，安宫牛黄丸降低了Th17细胞和转录因子RORγt的表达水平，增加了Treg细胞和转录因子Foxp3的表达水平。安宫牛黄丸降低了主动脉根部胶原纤维含量和炎性细胞浸润。总之，我们证明了安宫牛黄丸对高脂饮食诱导的ApoE小鼠早期和中期动脉粥样硬化模型具有抗动脉粥样硬化作用，通过调节Th17/Treg平衡、抑制慢性炎症、减少斑块胶原纤维和减少炎性细胞浸润，发挥其多途径多靶点抗早期和中期动脉粥样硬化的作用。
