Tinglu Yixin granule inhibited fibroblast-myofibroblast transdifferentiation to ameliorate myocardial fibrosis in diabetic mice

• Published in: Journal of ethnopharmacology 2025-01, Vol.337 (Pt 3), p.118980, Article 118980
• Main Authors: Zhang, Meng, Sun, Xuemei, Zhao, Fusen, Chen, Zhaoyang, Liu, Min, Wang, Pengqun, Lu, Pengyu, Wang, Xindong
• Format: Article
• Language: English
• Subjects: actin; Animals; cardiac output; cardiomyopathy; Cell Transdifferentiation - drug effects; collagen; Diabetes Mellitus, Experimental - complications; Diabetes Mellitus, Experimental - drug therapy; Diabetes Mellitus, Type 2 - complications; Diabetes Mellitus, Type 2 - drug therapy; Diabetic Cardiomyopathies - drug therapy; Diabetic myocardial fibrosis; Diet, High-Fat; Drugs, Chinese Herbal - pharmacology; echocardiography; family; fibroblasts; Fibroblasts - drug effects; Fibroblasts - metabolism; fibrosis; Fibrosis - drug therapy; fluorescent antibody technique; high fat diet; hypertrophy; immunohistochemistry; inflammation; intraperitoneal injection; islets of Langerhans; Male; mass spectrometry; Mice; Mice, Inbred C57BL; mitogen-activated protein kinase; Molecular Docking Simulation; muscles; Myocardium - metabolism; Myocardium - pathology; Myofibroblasts - drug effects; Myofibroblasts - metabolism; noninsulin-dependent diabetes mellitus; oxidative stress; pharmacology; phosphorylation; prognosis; protein synthesis; Streptozocin; streptozotocin; TGF-β1; Tinglu Yixin granule; traditional medicine; Transdifferentiation; transforming growth factor beta; transient receptor potential channels; TRPC6; ultra-performance liquid chromatography; Western blotting
• ISSN: 0378-8741; 1872-7573; 1872-7573
• DOI: 10.1016/j.jep.2024.118980

Myocardial fibrosis is one of the pathological characteristics of advanced diabetic cardiomyopathy (DCM) and serves as the strong evidence of poor prognosis. Among them, the transdifferentiation of cardiac fibroblasts (CFs) may play a crucial role in the development of myocardial fibrosis in DCM. Tinglu Yixin granule (TLYXG) has been clinically used for many years and can significantly improve cardiac function of patients with DCM. However, the effect of TLYXG on myocardial fibrosis in DCM remains unknown, and the underlying mechanisms of its efficacy have yet to be fully understood. This study aimed to investigate the impact and underlying mechanism of TLYXG on myocardial fibrosis in diabetes mice. The bioactive compounds in TLYXG were identified using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). The potential mechanism of TLYXG in treating DCM was predicted using network pharmacology combined with molecular docking and protein-protein docking. The mice model of type 2 diabetes were established by intraperitoneal injection of streptozotocin (STZ) and the high-fat diet (HFD). Indicators of pancreatic islet function, lipids, oxidative stress, and inflammatory factors were tested using kits. Cardiac function was assessed in diabetic mice using echocardiography. Histologic staining was performed to evaluate myocardial hypertrophy and fibrosis. Mechanistically, the hypothesis was tested through rescue experiments. The expression levels of transient receptor potential channel 6 (TRPC6), transforming growth factor-β1 (TGF-β1), collagen I (COL-I) and alpha-smooth muscle actin (α-SMA), along with the mRNA and phosphorylation levels of SMAD family member 3 (Smad3) and protein 38 mitogen-activated protein kinase (p38 MAPK), were assessed using quantitative RT-qPCR, Western blot, immunohistochemistry, and immunofluorescence. Neonatal lactating mice were used to extract primary CFs for vitro experiments. Scratch and transwell assays were conducted to assess CFs migration and invasion abilities. Western blot and immunofluorescence were used to evaluate the expression levels of CFs transdifferentiation markers COL-I and α-SMA. A total of 168 active ingredients were detected in TLYXG based on UPLC-MS and databases. Network pharmacology indicated that TLYXG could improve DCM through inflammatory mediator regulation of TRP channels, TGF-beta signaling pathway, and MAPK signaling pathway. ELISA results showed that TLYXG could ameliorate metabolic lev