School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China.
Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hong Kong, China.
Phytomedicine. 2019 Feb 15;54:182-194. doi: 10.1016/j.phymed.2018.09.021. Epub 2018 Sep 5.
Clinical evidence gathered in Chinese communities suggested that acupoint sticking therapy could be an alternative treatment for asthma-related diseases. However, its underlying mechanism is still poorly understood.
AIM/HYPOTHESIS: In this study, we aimed to investigate the mechanism of the anti-inflammatory effect of acupoint sticking application with 'Treatment of Winter Disease in Summer' (TWDS) prescription by using metabolomics.
Allergic asthma in guinea pig was sensitized and challenged by ovalbumin (OVA). Histopathological evaluation of the lung tissue was performed by hematoxylin and eosin (H&E) staining and Masson's trichrome staining. The levels of Th2 cytokine and IgE level in serum were measured using enzyme-linked immunoassay (ELISA). The mRNA expression levels of IL-4, IL-5, IL-13 and orosomucoid-like 3 (ORMDL3) were measured using quantitative reverse transcription polymerase chain reaction (RT-qPCR). Proteins of NF-κB signaling pathway were measured using western blot. The serum metabolomics profiles were obtained by using ultra-performance liquid chromatography combined with electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF-MS).
The overall results confirmed that AST with TWDS prescription had a significant protective effect against OVA-induced allergic asthma in guinea pig. This treatment not only attenuated airway inflammation and collagen deposition in the airway, but also decreased the levels of IL-4, IL-5, IL-13 and IgE in serum. In addition, metabolomics results indicated that metabolisms of phospholipid, sphingolipid, purine, amino acid and level of epinephrine were restored back to the normal control level. Moreover, results of the gene expression of ORMDL3 in lung tissues indicated that AST using TWDS could alter the sphingolipid metabolism. Further western blotting analysis also showed that its anti-inflammatory mechanism was by decreasing the phosphorylation of p65 and IκB.
The study demonstrated that metabolomics provides a better understanding of the actions of TWDS acupoint sticking therapy on OVA-induced allergic asthma.
在华人群体中收集的临床证据表明，穴位贴敷疗法可能是治疗哮喘相关疾病的一种替代疗法。然而，其潜在机制仍知之甚少。
目的/假设：本研究旨在通过代谢组学研究探讨“冬病夏治”穴位贴敷治疗的抗炎作用机制。
卵清蛋白（OVA）致敏和激发变应性哮喘豚鼠，苏木精-伊红（H&E）和 Masson 三色染色进行肺组织病理评估。酶联免疫吸附试验（ELISA）检测血清中 Th2 细胞因子和 IgE 水平。实时定量逆转录聚合酶链反应（RT-qPCR）检测 IL-4、IL-5、IL-13 和粘蛋白样 3（ORMDL3）的 mRNA 表达水平。Western blot 检测 NF-κB 信号通路蛋白。采用超高效液相色谱-电喷雾电离四极杆飞行时间质谱联用（UPLC-ESI-QTOF-MS）获得血清代谢组学图谱。
总体结果证实，AST 联合 TWDS 处方对 OVA 诱导的豚鼠变应性哮喘具有显著的保护作用。该治疗不仅减轻气道炎症和气道胶原沉积，还降低血清中 IL-4、IL-5、IL-13 和 IgE 水平。此外，代谢组学结果表明，磷脂、鞘脂、嘌呤、氨基酸代谢和肾上腺素水平得到恢复。此外，肺组织中 ORMDL3 的基因表达结果表明，AST 联合 TWDS 可改变鞘脂代谢。进一步的 Western blot 分析还表明，其抗炎机制是通过降低 p65 和 IκB 的磷酸化。
本研究表明，代谢组学为“冬病夏治”穴位贴敷疗法对 OVA 诱导的变应性哮喘的作用机制提供了更好的理解。
