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This study aimed to investigate the mechanism of the traditional Chinese medicine compound Shengmai Yin (SMY) in exerting anti-myocardial ischemia (MI) effects through the synergistic regulation of the gut microbiota and the MAPK signaling pathway. First, network pharmacology combined with GO and KEGG enrichment analysis was used to predict the potential targets and signaling pathways of SMY against MI. Secondly, an acute myocardial ischemia model was established in SD rats by intraperitoneal injection of pituitrin (25 U·kg-1), while SMY was administered by intragastric gavage for intervention for 7 days, with daily body weight recording. Subsequently, electrocardiogram and cardiac histopathological changes were detected. The levels of myocardial injury markers [aspartate aminotransferase (AST), creatine kinase (CK), lactate dehydrogenase (LDH), lactic acid (LD), and cardiac troponin-Ⅰ (cTn-Ⅰ)], oxidative stress indicators [superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA)] and inflammatory factors [tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β)] were measured. Immunohistochemistry was used to detect the expression of key apoptosis-related proteins in myocardial tissue, including Bcl-2 associated X protein (Bax), cysteinyl aspartate specific proteinase-3 (Caspase-3), and B-cell lymphoma-2 (Bcl-2). TUNEL staining was used to detect cardiomyocyte apoptosis. Western blot was used to detect the expression levels of key proteins in the mitogen-activated protein kinase (MAPK) signaling pathway. 16S rRNA sequencing was used to analyze the structural changes of the gut microbiota. The experimental animals were approved by the Experimental Animal Management Committee Ethics Committee of Chengdu University of Traditional Chinese Medicine (Approval No.: 2014 DL-023). Network pharmacology analysis identified pivotal targets including TP53, TNF, MAPK1, MAPK3, and CTNNB1, which are predominantly involved in the MAPK and PI3K-AKT signaling pathways. In vivo studies demonstrated that SMY ameliorated electrocardiographic abnormalities and histopathological lesions, reduced serum levels of myocardial injury biomarkers and pro-inflammatory cytokines, suppressed the phosphorylation of the key MAPK protein p38, downregulated the expression of Bax and Caspase-3, and upregulated Bcl-2. Furthermore, 16S rRNA sequencing revealed that SMY modulated gut microbiota diversity and compositional structure. These findings suggest that SMY exerts cardioprotective effects against myocardial ischemia, potentially by suppressing inflammatory responses and cardiomyocyte apoptosis. This therapeutic effect is associated with the modulation of the MAPK signaling pathway and alterations in gut microbiota composition; however, the precise mechanisms and causal relationships warrant further investigation.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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