Uncovering the molecular mechanisms of tonifying kidney and activating blood Decoction against myocardial fibrosis using network Pharmacology and experimental validation

Chronic heart failure(HF) has become a disease of global concern due to its high morbidity and mortality.This has highlighted the need for cardioprotective agents.The Tonifying Kidney and Activating Blood(KTBA) decoction has been approved for clinical treatment of chronic HF.Tanshinone IIA(Tan IIA), rooted from Salvia miltiorrhiza of KTBA, has been approved for treating cardiovascular conditions.However, the mechanism is still unclear.This study examined the impact of KTBA on cardiomyocyte fibrosis in a rat model of heart failure post-myocardial infarction, induced by ligation of the left anterior descending coronary artery, followed by exhaustive swimming and starvation. Additionally, the effects of Tan IIA on CCD-841CoN cells were assessed under ischemic conditions in a 37 °C incubator with hypoxic environment (1% O₂, 5% CO₂, and 94% N₂). The investigation employed an integrative approach combining network pharmacology with molecular mechanism analysis.The findings of network pharmacology indicate that KTBA may exert its influence by targeting key proteins such as TNF, AKT1, STAT3, RELA (NF-κB p65), NFκBIA (I-κBα), and MAPK14 (p38α).Results showed that KTBA increased SERCA2a level, lowered collagen I and III, α-SMA, and phospholamban levels, reduced collagen fiber deposition, and delayed mitochondria injury.This cardioprotection effect was perhaps due to suppressing the expressions of p38MAPK, I-κBα, NF-κB, AQP4,AKT, PI3K, TNF-α, and STAT3 and increasing the levels of ZO-1 and Occludin in hippocampus of chronic HF rats, which were partially diminished by SB203580 and PDTC.Additionally, Tan IIA reduced levels of p38MAPK, I-κBα, NF-κB, STAT3, and increased levels of AQP4, Claudin-1, ZO-1, and ZO-2,that were reduced by siRNAs targeting p38MAPK, NF-κB, and AQP4.In conclusion, by modulating the p38MAPK/NF-κB/AQP4 axis, KTBA decoction delays cardiomyocyte fibrosis through alleviating hippocampal blood-brain barrier and Tan IIA improves enterocyte barrier integrity.