# Study on the role of Shenfu injection in mediating ferroptosis through the Akt/GSK-3β/Nrf2 pathway in yang-deficient chronic heart failure

## TL;DR

This study explores how Shenfu injection improves heart function in rats with yang-deficient chronic heart failure by reducing ferroptosis through a specific signaling pathway.

## Contribution

The study reveals a novel mechanism by which Shenfu injection protects heart tissue via the Akt/GSK-3β/Nrf2 pathway in yang-deficient chronic heart failure.

## Key findings

- Shenfu injection improved cardiac function and reduced markers of heart damage in yang-deficient CHF rats.
- The treatment reduced ferroptosis markers and activated the Akt/GSK-3β/Nrf2 pathway in cardiac tissues.
- Blocking the Akt/GSK-3β/Nrf2 pathway reduced the protective effects of Shenfu injection.

## Abstract

The present study investigates the role of Shenfu injection in the treatment of yang-deficient chronic heart failure (CHF).

Sprague-Dawley (SD) rats were modeled for yang-deficient CHF by abdominal aortic coarctation. Echocardiography was performed to detect changes in cardiac function, and serum N-terminal B-type natriuretic peptide proteins (NT-proBNP), cardiac troponin I (cTnI), and ferroptosis-related factors were measured using ELISA kits. Pathological changes in cardiac tissues were observed through hematoxylin-eosin (HE) and Masson’ trichrome staining, cardiomyocyte apoptosis was measured by TUNEL staining, and reactive oxygen species (ROS) production was determined through dihydroethidium (DHE) staining. The expression of nuclear factor E2-related factor 2 (Nrf2), cyclooxygenase 2 (Ptgs2), glutathione peroxidase 4 (GPX4), solute carrier family 3 member 2 (SLC3A2), solute carrier family 7 member 11 (SLC7A11), and acyl-CoA synthetase long-chain family member 4 (ACSL4) in cardiac tissues were analyzed through RT-qPCR. Phosphorylated Akt (p-Akt), phosphorylated GSK-3β (p-GSK-3β), and Nrf2 expression in tissues were tested through immunohistochemistry. The protein expression of the Akt/GSK-3β/Nrf2 pathway was detected by Western blot. The Akt/GSK-3β/Nrf2 pathway inhibitor LY294002 was applied to the rats administrated with Shenfu injection.

Shenfu injection decreased the left ventricular end-diastolic diameter and left ventricular end-systole diameter and increased the left ventricular ejection fraction and left ventricular fractional shortening in rats with CHF. The treatment reduced NT-proBNP and cTnI levels, while improving pathological damage in the cardiac tissue. The treatment was also noted to decrease serum MDA, ACSL4, and Fe2+ and increase GSH, GPX4, SOD, and SLC3A2 in the sample; increase GPX4,SLC7A11 and SLC3A2 mRNA in cardiac tissues, and decrease Ptgs2 and ACSL4 mRNA. Shenfu injection was also noted to activate the Akt/GSK-3β/Nrf2 signaling pathway, while LY294002 weakened the therapeutic effect of the treatment on cardiac tissue damage.

Shenfu injection activates the Akt/GSK-3β/Nrf2 pathway to prevent myocardial injury and ferroptosis in yang-deficient CHF.