Abstract: BACKGROUND: Inflammaging, characterized by a chronic low-grade pro-inflammatory state during natural aging, is closely associated with the development of age-related diseases such as cardiovascular disorders. However, a systematic theoretical framework centered on cardiac inflammaging as a core pathological mechanism is still lacking. 
OBJECTIVE: To analyze the prescription patterns and medication principles for treating cardiac aging in traditional Chinese medicine (TCM) classics using data mining and network pharmacology methods, and to explore the potential mechanisms of core drugs. 
METHODS: (1) Formulations for treating cardiac aging were collected and compiled from the 2020 edition of the Pharmacopoeia of the People's Republic of China, the 10th edition of Chinese Materia Medica, and the Comprehensive Dictionary of TCM Prescriptions. The frequency of medicinal herbs, their properties, and meridian affinity were statistically analyzed, along with association rule analysis and clustering, to identify core herb pairs for treating cardiac aging. A core herb pair-active ingredient-disease-common target network model was established, followed by gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway analysis. In addition, molecular docking and dynamic simulation were employed to assess the binding ability and stability between potential key targets and active ingredients. (2) Validation experiment: Sixty C57BL/6 mice were randomly divided into three intervention groups: the blank group (n=20) was administered distilled water via gavage, the low-dose musk-aconite group (n=20) was administered 2.64 g/kg musk-aconite via gavage, and the high-dose musk-aconite group (n=20) was administered 5.28 g/kg musk-aconite group via gavage, once daily for 7 consecutive days. After the final administration, blood was collected from the orbital sinus, and serum was separated to obtain drug-free serum and low- and high-dose drug-containing serum of musk-aconite. Passage 3 human ventricular cardiomyocyte line AC16 was divided into three groups and treated with drug-free serum and low- and high-dose drug-containing serum of musk and aconite, respectively. Cell viability was detected using the cell counting kit-8 method. Passage 3 human ventricular cardiomyocyte line AC16 was divided into four groups and treated with PBS, drug-free serum, and low- and high-dose drug-containing serum of musk-aconite, respectively, for 48 hours. Then, tumor necrosis factor α was added to the latter three groups for 4 hours to induce an inflammatory model. Western blot was used to detect the expression of nuclear factor κB inhibitor protein, phosphorylated nuclear factor κB inhibitor protein, nuclear factor κB p65, and nuclear factor κB p50 protein.
RESULTS AND CONCLUSION: (1) A total of 268 prescriptions were identified from the 2020 edition of the Pharmacopoeia of the People's Republic of China. After removing duplicates and irrelevant prescriptions, 88 prescriptions containing 36 Chinese medicinal herbs were selected. The most frequently used herbs were cinnamon, dried ginger, aconite, shallot, and musk. These herbs were predominantly pungent and warm in nature, targeting the heart, kidney, and liver meridians, with a treatment principle of Fangxiang Wentong method (aromatic warm and unblocking method). Association rule analysis identified four strong herb pairings, with high support for "musk-aconite," "cinnamon-dried ginger," "dried ginger-aconite," and "shallot-trichosanthes" (confidence ≥ 90%). Factor analysis extracted four common factors, reflecting the historical TCM treatment philosophy of Fangxiang Wentong method. The four core herb pairs might alleviate cardiac aging by modulating inflammatory signaling pathways, which were correlated with the relatively high proportion of inflammatory markers in cardiac aging. Molecular docking of the "musk-aconite" pair revealed good binding ability and stability between key targets and their corresponding active ingredients. (2) Cell counting kit-8 assay results indicated that the "musk-aconite" pair exhibited high cellular compatibility with drug-containing serum. Western blot analysis revealed that compared with the control group, intervention with tumor necrosis factor α significantly induced phosphorylation of the nuclear factor κB inhibitor protein, leading to the release of the nuclear factor κB dimer (p65/p50) into the cell nucleus, thereby confirming the successful establishment of the inflammatory injury model. However, low- and high-dose musk-aconite drug pair-containing serum intervention significantly inhibited the upregulation of nuclear factor κB inhibitor protein and nuclear factor κB (p65/p50) in the cell nucleus induced by tumor necrosis factor α. To conclude, in traditional Chinese medicine, the treatment of cardiac aging primarily follows the principle of Fangxiang Wentong method, with the musk-aconite pair as the core. This herbal combination may slow the progression of cardiac aging by inhibiting inflammatory signals.

Key words: Fangxiang Wentong method, cardiac inflammaging, drug pairs, medication patterns, musk-aconite, inflammatory signal