The Second Hospital of Shandong University, Jinan, Shandong, China.
Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, Shandong, China.
Biomed Pharmacother. 2024 May;174:116548. doi: 10.1016/j.biopha.2024.116548. Epub 2024 Apr 9.
Various heart diseases ultimately lead to chronic heart failure (CHF). In CHF, the inflammatory response is associated with pyroptosis, which is mediated by the NOD-like receptor protein 3 (NLRP3) inflammasome. Fu Xin decoction (FXD) is commonly used in clinical practice to treat CHF and improve inflammatory conditions. However, the specific pharmacological mechanisms of action for FXD in these processes have yet to be fully understood.
The objective of this study was to examine the protective mechanism of FXT against CHF, both in H9c2 cells and mice.
A CHF mouse model was established, and the effect of FXD was observed via gavage. Cardiac function was evaluated using echocardiography, while serum BNP and LDH levels were analyzed to assess the severity of CHF. Hematoxylin and eosin staining (H&E) and Masson staining were performed to evaluate myocardial pathological changes, and TdT-mediated dUTP Nick-End Labeling staining was used to detect DNA damage. Additionally, doxorubicin was utilized to induce myocardial cell injury in H9c2 cells, establishing a relevant model. CCK8 was used to observe cell viability and detect LDH levels in the cell supernatant. Subsequently, the expression of pyroptosis-related proteins was detected using immunohistochemistry, immunofluorescence, and western blotting. Finally, the pharmacological mechanism of FXD against CHF was further validated by treating H9c2 cells with an NLRP3 activator and inducing NLRP3 overexpression.
According to current research findings, echocardiography demonstrated a significant improvement of cardiac function by FXD, accompanied by reduced levels of BNP and LDH, indicating the amelioration of cardiac injury in CHF mice. FXD exhibited the ability to diminish serum CRP and MCP inflammatory markers in CHF mice. The results of HE and Masson staining analyses revealed a significant reduction in pathological damage of the heart tissue following FXD treatment. The CCK8 assay demonstrated the ability of FXD to enhance H9c2 cell viability, improve cell morphology, decrease LDH levels in the cell supernatant, and alleviate cell damage. Immunohistochemistry, Western blotting, and immunofluorescence staining substantiated the inhibitory effect of FXD on the NLRP3/caspase-1/GSDMD pyroptosis signaling pathway in both CHF and H9c2 cell injury models. Ultimately, the administration of the NLRP3 activator (Nigericin) and the overexpression of NLRP3 counteract the effects of FXD on cardiac protection and pyroptosis inhibition in vitro.
FXD exhibits a cardioprotective effect, improving CHF and alleviating pyroptosis by inhibiting the NLRP3/caspase-1/GSDMD pathway.
各种心脏疾病最终都会导致慢性心力衰竭（CHF）。在 CHF 中，炎症反应与细胞焦亡有关，细胞焦亡是由 NOD 样受体蛋白 3（NLRP3）炎性小体介导的。复心汤（FXD）常用于临床治疗 CHF 和改善炎症状态。然而，FXD 在这些过程中的具体药理作用机制尚未完全阐明。
本研究旨在探讨 FXD 对 CHF 大鼠心肌细胞和小鼠的保护作用及机制。
建立 CHF 小鼠模型，通过灌胃观察 FXD 的作用。采用超声心动图评估心功能，分析血清 BNP 和 LDH 水平评估 CHF 严重程度。采用苏木精-伊红（H&E）和 Masson 染色评估心肌病理变化，TdT 介导的 dUTP 缺口末端标记（TUNEL）染色检测 DNA 损伤。此外，采用阿霉素诱导 H9c2 心肌细胞损伤，建立相关模型。CCK8 法观察细胞活力，检测细胞上清液中 LDH 水平。随后，采用免疫组化、免疫荧光和 Western blot 检测焦亡相关蛋白的表达。最后，通过用 NLRP3 激活剂处理 H9c2 细胞并诱导 NLRP3 过表达，进一步验证 FXD 治疗 CHF 的药理学机制。
根据目前的研究结果，FXD 可显著改善 CHF 小鼠的心脏功能，降低 BNP 和 LDH 水平，表明 FXD 可改善 CHF 小鼠的心脏损伤。FXD 可降低 CHF 小鼠血清 CRP 和 MCP 炎症标志物水平。HE 和 Masson 染色分析结果显示，FXD 治疗后心脏组织病理损伤明显减轻。CCK8 检测结果表明，FXD 可增强 H9c2 细胞活力，改善细胞形态，降低细胞上清液中 LDH 水平，减轻细胞损伤。免疫组化、Western blot 和免疫荧光染色证实，FXD 可抑制 CHF 和 H9c2 细胞损伤模型中的 NLRP3/caspase-1/GSDMD 焦亡信号通路。最终，给予 NLRP3 激活剂（尼日尼亚菌素）和过表达 NLRP3 可抵消 FXD 在体外对心脏保护和抑制焦亡的作用。
FXD 通过抑制 NLRP3/caspase-1/GSDMD 通路发挥心脏保护作用，改善 CHF 并减轻细胞焦亡。
