Chen Jinhong, Liu Zhichao, Deng Fangjun, Liang Jiayu, Fan Boya, Zhen Xin, Tao Rui, Sun Lili, Zhang Shaoqiang, Cong Zidong, Li Xiaofeng, Du Wuxun
Graduate School, Tianjin University of TCM, Tianjin 301617, China.
Department of TCM, Tianjin University of TCM, Tianjin, 301617, China.
Phytomedicine. 2022 May;99:153989. doi: 10.1016/j.phymed.2022.153989. Epub 2022 Feb 12.
Lian-Gui-Ning-Xin-Tang (LGNXT), a classical traditional Chinese medicine (TCM) formula, has been widely used in clinical practice and has shown satisfactory efficacy in the treatment of arrhythmias. However, its mechanism of action in the treatment of arrhythmias is still unknown. Moreover, the complex chemical composition and therapeutic targets of LGNXT pose a challenge in pharmacological research.
To analyze the active compounds and action mechanisms of LGNXT for the treatment of arrhythmias.
Here, we used an integrated pharmacology approach to identify the potential active compounds and mechanisms of action of LGNXT in treating arrhythmias. Potential active compounds in LGNXT were identified using ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS) and the potential related targets of these compounds were predicted using an integrated in silico approach. The obtained targets were mapped onto relevant databases to identify their corresponding pathways, following the experiments that were conducted to confirm whether the presumptive results of systemic pharmacology were correct.
Eighty-three components were identified in herbal materials and in animal plasma using UPLC-Q-TOF/MS and were considered the potential active components of LGNXT. Thirty key targets and 57 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were identified as possible targets and pathways involved in LGNXT-mediated treatment using network pharmacology, with the cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA)/Ca system pathway being the most significantly affected. This finding was validated using an adrenaline (Adr)-induced rat model of arrhythmias. Pretreatment with LGNXT delayed the occurrence, shortened the duration, and reduced the severity of arrhythmias. LGNXT exerted antiarrhythmic effects by inhibiting cAMP, PKA, CACNA1C, and RyR2.
The findings of this study revealed that preventing intracellular Ca overload and maintaining intracellular Ca homeostasis may be the primary mechanisms of LGNXT in alleviating arrhythmias. Thus, we suggest that the β-adrenergic receptor (AR)/cAMP/PKA/Ca system signaling hub may constitute a promising molecular target for the development of novel antiarrhythmic therapeutic interventions. Additionally, we believe that the approach of investigation of the biological effects of a multi-herbal formula by the combination of metabolomics and network pharmacology, as used in this study, could serve as a systematic model for TCM research.
连归宁心汤（LGNXT）是一种经典的中药方剂，已在临床实践中广泛应用，并且在治疗心律失常方面显示出令人满意的疗效。然而，其治疗心律失常的作用机制仍不清楚。此外，LGNXT复杂的化学成分和治疗靶点给药理学研究带来了挑战。
分析LGNXT治疗心律失常的活性成分及作用机制。
在此，我们采用综合药理学方法来确定LGNXT治疗心律失常的潜在活性成分及作用机制。使用超高效液相色谱-四极杆-飞行时间质谱（UPLC-Q-TOF/MS）鉴定LGNXT中的潜在活性成分，并使用综合的计算机方法预测这些化合物的潜在相关靶点。将获得的靶点映射到相关数据库以确定其相应途径，随后进行实验以确认系统药理学的推测结果是否正确。
使用UPLC-Q-TOF/MS在药材和动物血浆中鉴定出83种成分，并将其视为LGNXT的潜在活性成分。使用网络药理学鉴定出30个关键靶点和57条京都基因与基因组百科全书（KEGG）途径，作为LGNXT介导治疗中可能涉及的靶点和途径，其中环磷酸腺苷（cAMP）/蛋白激酶A（PKA）/钙系统途径受影响最为显著。这一发现通过肾上腺素（Adr）诱导的大鼠心律失常模型得到验证。LGNXT预处理可延迟心律失常的发生、缩短其持续时间并降低其严重程度。LGNXT通过抑制cAMP、PKA、CACNA1C和RyR2发挥抗心律失常作用。
本研究结果表明，防止细胞内钙超载和维持细胞内钙稳态可能是LGNXT缓解心律失常的主要机制。因此，我们认为β-肾上腺素能受体（AR）/cAMP/PKA/钙系统信号枢纽可能是开发新型抗心律失常治疗干预措施的一个有前景的分子靶点。此外，我们相信本研究中使用的代谢组学和网络药理学相结合来研究多草药配方生物学效应的方法，可作为中医药研究的一个系统模型。
