Enhanced lipid metabolism reprogramming in CHF rats through IL-6-mediated cardiac glial cell modulation by digilanid C and electroacupuncture stimulation combination

Abstract

Background: Cardiac lipid metabolism reprogramming is recognized as a critical pathological factor in the progression of chronic heart failure (CHF). The therapeutic potential of digilanid C and electroacupuncture stimulation (ES) in enhancing lipid metabolism and cardiac function has been established. However, the optimal synergistic regulatory strategies of these interventions on cardiac lipid metabolism have yet to be elucidated.

Methods: This study aimed to comprehensively evaluate the impact of a digilanid C-ES combination on cardiac steatosis remodeling in CHF. Assessments were conducted across various dimensions, including myocardial oxygen consumption, mitochondrial function, and lipid metabolism. Additionally, we sought to uncover the underlying neuromolecular mechanisms.

Results: Our findings, at both molecular and morphological levels, indicated that the synergistic application of digilanid C and ES significantly inhibited myocardial fibrosis and steatosis. This combination therapy facilitated the repair of cardiac neuro-vascular uncoupling and induced a reprogramming of lipid metabolism. Notably, the digilanid C-ES combination ameliorated cardiomyocyte apoptosis and enhanced mitochondrial biogenesis in CHF, leading to a restructured energy supply pattern. Cardiac immunofluorescence analyses revealed the aggregation of cardiac glial cells (CGCs) at sites of abnormal neurovascular coupling, a response to cardiac lipid degeneration. This was accompanied by a marked reduction in the abnormally elevated expression of interleukin 6 (IL-6) and glutamatergic signaling, which correlated with the severity of cardiac steatosis and the aberrant activation of CGCs. The combined therapy was found to activate the Janus kinase 1 (JAK1)/signal transducer and activator of transcription 3 (STAT3) pathway, effectively attenuated lipid accumulation and over-recruitment of CGCs and deprivation of glutamatergic nerves.

Conclusion: These findings underscore the potential of digilanid C and ES combination therapy as a novel approach to modulate the complex interplay between neurovascular dynamics and metabolic dysregulation in CHF.

Keywords: cardiac glial cells; chronic heart failure; digilanid C-electroacupuncture stimulation; glutamatergic; lipid.

FIGURE 1

Digilanid C-ES combination effectively improves cardiac function and cardiac energy supply in rats with CHF. (A) Representative images of HE and (B) Masson staining of rat hearts in each group (×200 magnification). (C) The modulation advantage of digilanid C-ES combination on serum BNP in CHF rats. (D) Differences in LVEF and (E) representative images of cardiac ultrasound in LVEF in each group of rats. (F) Effect of sildenafil injection on myocardial oxygen consumption in rats after electroacupuncture intervention. (H) Adverse event statistics for each intervention group. ^# p < 0.05 vs. the normal control, ^* p < 0.05 vs. the model or another treatment group. ES, electracupuncture stimulation; LVEF, left ventricular injection fraction; BNP, brain natriuretic peptide; RPP, rate pressure product; HR, heart rate; SBP, systolic pressure.

FIGURE 2

Digilanid C-ES combination to regulate cardiac fat overload. Typical representative plots of ORO staining of the liver in the (A1) normal, (A2) model, (A3) digilanid C, (A4) ES, and (A5) digilanid C-ES combination groups. Differences in the degree of fatty degeneration in kidney and (C) heart tissues in different groups. (D) Modulation of pericardial fat wet weight by different intervention groups. (E) Correlation analysis between liver wet weight and pericardial fat wet weight. (F, G) Effect on peroxisome proliferators activated recepotor-γ (or PPAR-γ), (F, H) fatty acid binding protein 4 (FABP4), (F, I) hormone-sensitive triglyceride lipase (HSL) expression in heart. Vinculin was used as an internal reference protein. (J) Correlation between FABP4 and TRPV1 as indicated by quantitative analysis of co-localization immunofluorescence. ^# p < 0.05 vs. the normal control, ^* p < 0.05 vs. the model or another treatment group. (K) Representative images of FABP4 and CD31 immunofluorescence co-expression in heart (×200 magnification). DAPI stained the nuclei (blue), the red immunofluorescence represents the FABP4 and the green immunofluorescence represents the CD31. ES, electracupuncture stimulation; DAPI, 4′,6-diamidino-2-phenylindole.

FIGURE 3

Digilanid C-ES combination to remodeling cardiac energy metabolism. (A, B) Effect on chemokine 2 (CCL2), (A, C) peroxisome proliferator-activated-recptor-γ coactivator-1α (PCG-1α), (A, D) mitochondrial transcription factor A (TFAM) expression in heart. Vinculin was used as an internal reference protein. (E) Quantitative fluorescence analysis of NKA-positive cells. (F) Representative images of NKA immunofluorescence in pancreas of model group (×200 magnification). DAPI stained the nuclei (blue) and the red immunofluorescence represents the NKA. (G) Representative images of FABP4 and NKA immunofluorescence co-expression in heart of model group (×200 magnification). DAPI stained the nuclei (blue), the red immunofluorescence represents the NKA and the green immunofluorescence represents the FABP4. (H) Correlation between FABP4 and NKA as indicated by quantitative analysis of co-localization immunofluorescence. ^# p < 0.05 vs. the normal control, ^* p < 0.05 vs. the model or another treatment group. ES, electracupuncture stimulation; NKA, sodium-potassium pump; DAPI, 4′,6-diamidino-2-phenylindole; FABP4, fatty acid binding protein 4 3.

FIGURE 4

Digilanid C-ES combination drived cardiac glial cells to remodel cardiac neuro-vascular coupling. (A, D) Differences in immunofluorescence co-localization of PGP9.5 and CD31 between groups. (B, E) Differences in the response of CD31 and TH under different intervention conditions. (C, F) Effect on GFAP and TH co-expression after needle-medicine combination intervention. (G, H) Effect on bcl-2, (G, I) beclin one expression in heart. Vinculin was used as an internal reference protein. ^# p < 0.05 vs. the normal control, ^* p < 0.05 vs. the model or another treatment group. DAPI, 4′,6-diamidino-2-phenylindole; ES, electracupuncture stimulation; PGP9.5, proteingene product 9.5; CD31, or platelet endothelial cell adhesion molecule-1; TH, tyrosine hydroxylase; GFAP, glial fibrillary acidic protein.

FIGURE 5

Digilanid C-ES combination harmonizes cardiac glial cell-glutamatergic fibers interactions to ameliorate cardiac lipotoxicity. (A, B) Effect on Janus kinase 1 (JAK1), (A, C) signal transducer and activator of transcription 3 (STAT3), (A, D) cJun and (A, E) interleukin 6 (IL-6) expression in heart. Vinculin was used as an internal reference protein. (F, H) Differences in immunofluorescence co-localization of IL-6 and GFAP between groups. (G, I) Differences in the response of GFAP and VGLUT1 under different intervention conditions. (J, K) Effect on GFAP and IL-6 co-expression after needle-medicine combination intervention. ^# p < 0.05 vs. the normal control, ^* p < 0.05 vs. the model or another treatment group. DAPI, 4′,6-diamidino-2-phenylindole; ES, electroacupuncture stimulation; PGP9.5, proteingene product 9.5; CD31, or platelet endothelial cell adhesion molecule-1; TH, tyrosine hydroxylase; GFAP, glial fibrillary acidic protein; VGLUT1, vesicular glutamate transporter one.