Therapeutic effect of Fuzheng Yixin decoction (扶正益心方) on cardiac insufficiency via the soluble guanylyl cyclase-cyclic guanosine monophosphate-protein kinase G pathway in a doxorubicin-induced dilated cardiomyopathy rat model

doi:10.19852/j.cnki.jtcm.20260514.002

Abstract

Abstract:

OBJECTIVE: To investigate the effect of Fuzheng Yixin decoction (扶正益心方, FZYX) and explore its underlying mechanisms in doxorubicin-induced dilated cardiomyopathy (DCM) rat model.

METHODS: Sprague-Dawley rats were intraperitoneally injected with doxorubicin for 6 consecutive weeks. Ultra-high-perform ance liquid chromatography Q-exactive mass spectrometry was utilized to obtain mass spectrum data for FZYX. The rats were then intragastrically injected with low-dose FZYX, high-dose FZYX, or captopril for 4 weeks. Serum N-terminal pro-brain natriuretic peptide, transthoracic echocardiography, haematoxylin and eosin and Masson staining, electron microscopy, and immunohistochemistry were used to evaluate the efficacy of FZYX. Differentially expressed genes and metabolites in the myocardial tissue of DCM rats following FZYX intervention were subsequently identified through transcriptomics and non-targeted metabolomics. Finally, the influence of FZYX on the expression of natriuretic peptide precursor A (Nppa), natriuretic peptide precursor B (Nppb), soluble guanylyl cyclase (sGC), protein kinase G (PKG), and PKG-1α in DCM rats was verified through quantitative polymerase chain reaction (qPCR), Western blotting (WB) based on the results of transcriptomics and non-targeted metabolomics analyses.

RESULTS: FZYX alleviated body weight loss; inhibited ventricular dilatation; restored impaired heart function, ruptured sarcomeres and mitochondrial damage, and pathological changes; and reduced collagen deposition. Transcriptomic and non-targeted metabolomic results revealed that FZYX regulated the expression of sGC-cyclic guanosine monophosphate (cGMP)-PKG pathway-related genes (Nppa, Nppb) and metabolites such as cGMP, 5'-guanylic acid, adenosine. qPCR and WB demonstrated that FZYX suppressed the expression of Nppa and Nppb and increased the expression of sGC, which led to a further increase in the level of PKG and PKG-1α.

CONCLUSION: These findings suggest that the cardioprotective effect of FZYX may be mediated through the restoration of the sGC-cGMP-PKG signaling pathway. FZYX may provide a new strategy for clinical application against DCM.

Key words: dilated cardiomyopathy, transcriptomics, non-targeted metabolomics, soluble guanylyl cyclase-cyclic guanosine monophosphate-protein kinase G pathway

SANG Xiuxiu, Lü Shuquan, WANG Feng, YANG Jing, WANG Qingguo, WANG Lixin. Therapeutic effect of Fuzheng Yixin decoction (扶正益心方) on cardiac insufficiency via the soluble guanylyl cyclase-cyclic guanosine monophosphate-protein kinase G pathway in a doxorubicin-induced dilated cardiomyopathy rat model[J]. Journal of Traditional Chinese Medicine, 2026, 46(4): 854-863.

Figures/Tables 5

Table 1 Serum NT-proBNP, and the results of transthoracic echocardiography ($\bar{x}±s$)

Group	n	NT-proBNP (pg/mL)	LVEF (%)	LVEDD (mm)	LVESD (mm)	LVFS (%)	IVST (mm)	LVPWT (mm)
Con	8	616.68±136.63	72.30±3.87	7.33±0.43	4.18±0.35	42.83±3.40	3.37±0.22	2.24±0.43
Dox	7	903.36±142.19^a	62.10±5.90^a	9.01±0.61^a	5.82±0.16^a	35.20±4.52^a	2.57±0.47^a	1.71±0.18^a
FZYX-L	6	663.17±105.73^b	64.00±3.88	8.28±0.38	5.09±0.33^b	36.25±2.97	2.91±0.23	2.14±0.34
FZYX-H	7	633.57±135.64^b	71.99±3.90^b	7.93±0.50^b	4.56±0.45^b	42.79±3.40^b	3.13±0.31	2.32±0.44^b
Cap	7	636.29±128.73^b	70.84±7.06^b	8.15±0.25^b	4.59±0.69^b	42.05±6.43^b	3.13±0.62	1.80±0.24

Figure 1 FZYX restricts myocardial pathological injury and mitochondrial damage A: cardiac chambers of rats stained by HE, n = 1 for each group; B: left ventricle myocardial pathological staining by HE, magnification × 200, n = 3 for each group; C: left ventricle myocardial transmission electron microscope, magnification × 25 000, n = 3 for each group. A1, B1, C1: Control group; A2, B2, C2: Dox group; A3, B3, C3: FZYX-L group; A4, B4, C4: FZYX-H group; A5, B5, C5: Cap group. Con group: intragastric injected with saline; Dox group: intragastric injected with saline; FZYX-L: intragastric injected with 15 g/kg FZYX; FZYX-H: intragastric injected with 30 g/kg FZYX; Cap: intragastric injected with 5.0 mg/kg captopril. FZYX: Fuzheng Yixin decoction; Con: Control group; HE: haematoxylin and eosin; Dox: doxorubicin.

Figure 2 FZYX attenuates myocardial fibrosis in DCM rats A: Masson staining (× 200); B: immunohistochemistry for α-SMA (× 200); C: immunohistochemistry for Col I (× 200); D: immunohistochemistry for Col Ⅲ (× 200); A1, B1, C1, D1: Control group; A2, B2, C2, D2: Dox group; A3, B3, C3, D3: FZYX-L group; A4, B4, C4, D4: FZYX-H group, A5, B5, C5, D5: Cap group. Con group: intragastric injected with saline; Dox group: intragastric injected with saline; FZYX-L: intragastric injected with 15 g/kg FZYX; FZYX-H: intragastric injected with 30 g/kg FZYX; Cap: intragastric injected with 5.0 mg/kg captopril. n = 3 for each group. FZYX: Fuzheng Yixin decoction; Con: Control group; HE: haematoxylin and eosin; Dox: doxorubicin; CVF: collagen volume fraction; α-SMA: alpha-smooth muscle actin; Col Ⅰ: collagen Ⅰ; Col Ⅲ: collagen Ⅲ.

Table 2 FZYX attenuates myocardial fibrosis in DCM rats (%, $\bar{x}±s$)

Group	n	CVF	α-SMA positive	Col I positiv	Col III positive
Con	3	4.74±2.08	0.99±0.53	3.18±1.45	7.67±2.07
Dox	3	25.87±6.09^a	2.99±0.47^a	11.157±1.75^a	14.93±1.49^a
FZYX-L	3	11.76±1.76^b	2.01±0.48	7.05±1.57^b	9.39±2.46^b
FZYX-H	3	6.01±2.67^b	0.64±0.22^b	5.48±1.49^b	7.24±1.79^b
Cap	3	13.28±4.10^b	1.55±0.09^b	3.04±0.71^b	5.49±1.99^b

Figure 3 FZYX activates sGC-cGMP-PKG signaling pathway in DCM rats A: the Nppa mRNA expression validated by qPCR; B: the Nppb mRNA expression validated by qPCR; C: the mRNA expression of sGCβ was measured by qPCR; D: the level of serum NO; E: protein isolated from myocardial tissues were analyzed by Western blot; F: the statistical analysis of WB; G: the statistical analysis of WB. Con group: intragastric injected with saline; Dox group: intragastric injected with saline; FZYX-H: intragastric injected with 30 g/kg FZYX Nppa: natriuretic peptide precursor A; Nppb: natriuretic peptide precursor B; sGC: soluble guanylyl cyclase; PKG: protein kinase G; PKG-1α: protein kinase G-1α; qPCR: quantitative polymerase chain reaction. Data is expressed as mean ± standard deviation (n = 3). aP < 0.05, compared with the control group; bP < 0.05, compared with the Dox group.

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