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基于巨噬细胞极化探究清热化痰活血方抗动脉粥样硬化的作用机制
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| 篇名： | 基于巨噬细胞极化探究清热化痰活血方抗动脉粥样硬化的作用机制 |
| TITLE： | Mechanism of action of Qingre huatan huoxue decoction against atherosclerosis based on macrophage polarization |
| 摘要： | 目的 基于巨噬细胞极化探究清热化痰活血方抗动脉粥样硬化（AS）的作用机制。方法制备清热化痰活血方含药血清，以阿托伐他汀为阳性对照，用以处理RAW264.7巨噬细胞；检测巨噬细胞活性、凋亡率和CD86、CD206的荧光强度以及肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）水平。将喂食高脂饲料的载脂蛋白e基因缺乏（ApoE－/－）小鼠（AS模型小鼠）随机分为模型组、阿托伐他汀组（2.6mg/kg）及清热化痰活血方低、中、高剂量组（90、180、360mg/kg），另将喂食普通饲料的C57BL/6J小鼠作为正常对照组，每组10只；给药组小鼠灌胃相应药物，每日1次，连续8周。检测各组小鼠血清中TNF-α、IL-1β水平，观察主动脉脂质沉积情况（以主动脉整体及根部斑块占比计）和主动脉根部的形态学变化，检测主动脉组织中CD86、CD206表达水平及诱导型一氧化氮合酶（iNOS）、精氨酸酶1（Arg-1）、AMP激活蛋白激酶（AMPK）、磷酸化AMPK（p-AMPK）、过氧化物酶体增殖物激活受体（PPAR-γ）蛋白表达水平。结果细胞实验结果显示，5～100μg/mL清热化痰活血方含药血清能显著提高RAW264.7细胞活性（P＜0.05），10、50、100μg/mL清热化痰活血方含药血清和阿托伐他汀能显著降低RAW264.7细胞凋亡率、CD86荧光强度和TNF-α、IL-1β水平，并显著升高CD206荧光强度（P＜0.05）。动物实验结果显示，与模型组比较，清热化痰活血方各剂量组和阿托伐他汀组小鼠血清中TNF-α、IL-1β水平，主动脉整体及根部斑块占比，CD86表达和iNOS蛋白表达均显著降低/下调，而CD206表达和Arg-1、PPAR-γ蛋白表达以及AMPK磷酸化表达均显著上调（P＜0.05）；主动脉病理形态均有不同程度改善。结论清热化痰活血方可通过调控巨噬细胞极化，增加M2型巨噬细胞比例，进而有效抑制AS斑块形成、减轻炎症反应，发挥抗AS的作用。 |
| ABSTRACT： | OBJECTIVE To explore the mechanism of action of Qingre huatan huoxue decoction against atherosclerosis （AS）based on macrophage polarization. METHODS Using atorvastatin served as the positive control， the drug-containing serum of the Qingre huatan huoxue decoction was prepared to treat RAW264.7 macrophages. Macrophage viability， apoptosis rate， and the fluorescence intensities of CD86 and CD206 were measured， along with the levels of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β）. Apolipoprotei n E-deficient （ApoE －/－ ） mice （AS model mice） fed with a high-fat diet were randomly assigned to model group， atorvastatin group （2.6 mg/kg）， and low-， medium- and high-dose groups of Qingre huatan huoxue decoction （90， 180， 360 mg/kg）， respectively. C57BL/6J mice fed with a standard diet served as the normal control group， with 10 mice per group. The treatment group mice were administered the corresponding drugs intragastrically， once daily， for 8 consecutive weeks. Serum levels of TNF-α and IL-1β were measured in all groups. Lipid deposition in the aorta （assessed by the percentage of plaque in the entire aorta and aortic root） and morphological changes in the aortic root were observed. Expression levels of CD86 and CD206 in aortic tissue， as well as the protein expression levels of inducible nitric oxide synthase （iNOS）， arginase-1 （Arg-1）， AMP-activated protein kinase （AMPK）， phosphorylated AMPK （p-AMPK）， and peroxisome proliferator-activated receptor γ （PPAR-γ） in aortic tissues were all detected. RESULTS Cell experiment results showed that， at concentrations of 5-100 μg/mL， the drug-containing serum of the Qingre huatan huoxue decoction significantly increased RAW264.7 cell viability （ P ＜0.05）. The drug-containing serum of the Qingre huatan huoxue decoction at concentrations of 10， 50， and 100 μg/mL， along with atorvastatin， significantly reduced apoptosis rates， CD86 fluorescence intensity， and TNF-α and IL-1β levels in RAW264.7 cells， while markedly enhancing CD206 fluorescence intensity （ P ＜0.05）. Animal experiment results showed that， compared with the model group， all dosage groups of Qingre huatan huoxue decoction and the atorvastatin group showed significantly reduced/down-regulated levels of TNF-α and IL-1β in serum， along with decreased aortic total and root plaque percentages， CD86 expression， and iNOS protein expression. CD206 expression and Arg-1， p-AMPK/AMPK， PPAR-γ protein expression were significantly up-regulated （ P ＜0.05）. Pathological morphology of the aorta showed varying degrees of improvement. CONCLUSIONS The formula of Qingre huatan huoxue decoction exerts its anti-AS effects by regulating macrophage polarization， increasing the proportion of M2 macrophages， thereby effectively inhibiting AS plaque formation and reducing inflammatory responses. |
| 期刊： | 2026年第37卷第04期 |
| 作者： | 钟华平；朱启城；邹征伟；何正义；谢和平；陈旭；段志胜；肖天 |
| AUTHORS： | ZHONG Huaping， ZHU Qicheng，ZOU Zhengwei，HE Zhengyi，XIE Heping，CHEN Xu，DUAN Zhisheng，XIAO Tian |
| 关键字： | 清热化痰活血方；巨噬细胞极化；动脉粥样硬化；AMPK/PPAR-γ信号通路 |
| KEYWORDS： | Qingre huatan huoxue decoction； macrophage polarization； atherosclerosis； AMPK/PPAR-γ signaling pathway |
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