Shenxiang Suhe pill improves cardiac function through modulating gut microbiota and serum metabolites in rats after acute myocardial infarction

Figures & data

Table 1. Diagrammatic representation of groups.

Group	Intervention	Dose
Sham	Saline fed	–
Model	Saline fed	–
Ato	Atorvastatin fed	20 mg/kg
Low	SXSH fed	0.063 g/kg
Medium	SXSH fed	0.126 g/kg
High	SXSH fed	0.252 g/kg
Model-ABX	Antibiotics fed	62.5 µg/mL ampicillin, 62.5 µg/mL metronidazole, 62.5 µg/mL neomycin and 31.25 µg/mL vancomycin
SXSH-ABX	SXSH and Antibiotics fed	0.126 g/kg SXSH, 62.5 µg/mL ampicillin, 62.5 µg/mL metronidazole, 62.5 µg/mL neomycin and 31.25 µg/mL vancomycin

Figure 1. SXSH protects cardiac function after AMI. (A) The schematic diagram for SXSH administration in rats after AMI. (B) Representative parasternal long-axis views and M-mode images. (C) Representative color Doppler imaging of the cardiac outflow tract and blood flow velocity. (D-F) The quantization charts corresponding to EF, FS, and AV. (G) Representative heart pictures of H&E staining (scale bar equals 100 µm). serum activities of (H) LDH, and (I) CK-MB. ^#p < 0.05, ^##p < 0.01 vs. Sham and *p < 0.05, **p < 0.01 vs. Model.

Figure 2. SXSH modulates gut microbiota post AMI. (A) Alpha diversity of Shannon index in each group. (B) Beta diversity of Bray Curtis-based PCoA of medium dose SXSH groups for 3 days, 7 days, and 14 days administration. (C) Statistical analysis of representative OTUs among different groups after 14 days administration.

Table 2. Changes in gut microbiota on different days.

OTU	Bacteria	3 Day		7 Day		14 Day
		S-M	D-M	S-M	D-M	S-M	D-M
OTU_3	s__Escherichia_coli	+	+	+	+
OTU_7	s__Lactobacillus_intestinalis	+	+				+
OTU_9	g__Allobaculum	+		+			+
OTU_24	g__Romboutsia	+				−
OTU_6	f__Muribaculaceae	+	+	+	−	−
OTU_1	s__Lactobacillus_gasseri	−	−	+	+	+	+
OTU_26	f__Muribaculaceae	−
OTU_17	s__Enterococcus_durans	−	−	+
OTU_4	g__unidentified_Prevotellaceae	−	−
OTU_12	f__Muribaculaceae	−	−
OTU_2	g__Lactobacillus		+		+	−	−
OTU_10	f__Muribaculaceae		+	−
OTU_58	g__Bacteroides		−
OTU_11	f__Muribaculaceae			−	−
OTU_8	f__Muribaculaceae			−			+
OTU_43	f__Ruminococcaceae			−	−
OTU_22	s__Akkermansia_muciniphila			−
OTU_29	f__Lachnospiraceae				+	+
OTU_18	f__Prevotellaceae				+	+
OTU_25	g__unidentified_Ruminococcaceae				−
OTU_27	f__Muribaculaceae				−
OTU_20	f__Prevotellaceae					+
OTU_13	f__Muribaculaceae					+
OTU_5	s__Lactobacillus_reuteri					−	−
OTU_55	f__Ruminococcaceae					−
OTU_15	o__Bacteroidales						+
OTU_23	f__Prevotellaceae						−
OTU_39	f__Muribaculaceae						−
OTU_84	f__Muribaculaceae						−

“S-M” represents Sham vs. Model; “D-M” represents SXSH administration group vs. Model; “+” represents top 5 upregulated bacteria; “−” represents top 5  downregulated bacteria. 

Figure 3. Antibiotics induced dysbacteriosis interferes cardiac protective effect of SXSH. (A) The schematic diagram for SXSH and ABX administration in AMI rats. (B) Representative parasternal long-axis views and M-mode images, and color Doppler imaging of the cardiac outflow tract and blood flow velocity. (C-E) The quantization charts corresponding to EF, FS, and AV. (F) Representative heart pictures of H&E staining [scale bar equals 3000 µm (top), 650 µm and 75 µm (bottom)]. serum activities of (G) SOD, (H) MDA, (I) LDH, and (J) CK-MB. ^#p < 0.05, ^##p < 0.01, ^###p < 0.001 vs. Sham, **p < 0.01, ***p < 0.001 vs. Model, and ^{^^}p < 0.01, ^{^^^}p < 0.001 vs. SXSH.

Figure 4. Circulating metabolomics for the quantification of metabolites among different groups. (A) The PCA score plots between Sham and Model in positive ion mode [R²Xp1 = 0.336, R²Xp2 = 0.224] and negative ion mode [R²Xp1 = 0.327, R²Xp2 = 0.267]. (B) The OPLS-DA score plots between Sham and Model in positive ion mode [R²X = 0.422, R²Y = 0.973, Q² = 0.859] and negative ion mode [R²X = 0.346, R²Y = 0.912, Q² = 0.695]. (C) Volcano plot showing the differentially accumulated and significantly changed metabolites in Model group compared to Sham. (D) The PCA score plots between Sham, Model, and SXSH in positive ion mode [R²Xp1 = 0.21, R²Xp2 = 0.149, R²Xp3 = 0.116, R²Xp4 = 0.112] and negative ion mode [R²Xp1 = 0.229, R²Xp2 = 0.182, R²Xp3 = 0.119]. (E) Pathway enrichment based on altered metabolites.

Table 3. SXSH administration regulates differential metabolites

Number	Metabolite	M/Z	Formula	Retention time
1	2-Nonen-1-ol	302.3	C9H18O	6.02135	↓
2	d-Pipecolic acid	130.0	C6H11NO2	0.479767	↓
3	THTC	150.0	C5H8O2S	0.77105	↓
4	LysoPC(P-18:0)	508.3	C26H54NO6P	6.56685	↓
5	Enantio-PAF C-16	524.3	C26H54NO7P	6.589167	↓
6	PC(16:0/20:4(5Z, 8Z, 11Z, 14Z))	804.5	C44H80NO8P	7.334717	↑
7	ent-7-Oxo-8(14), 15-pimaradien-19-oic acid	361.1	C20H28O3	4.348117	↑
8	N, N-Dimethyl-Safingol	330.3	C20H43NO2	6.0664	↓
9	Choline	104.1	C5H13NO	6.626383	↓
10	LysoPC(20:2(11Z, 14Z))	530.3	C28H54NO7P	6.626383	↓
11	LysoPE(22:0/0:0)	560.3	C27H56NO7P	6.6935	↓
12	PC(19:0/0:0)	538.3	C27H56NO7P	6.700983	↓
13	PC(22:1(11Z)/0:0)	578.4	C30H60NO7P	6.783167	↓
14	PS(DiMe(11, 3)/DiMe(13, 5))	968.5	C50H86NO12P	7.665267	↑
15	LysoPC(20:0)	552.4	C28H58NO7P	6.805583	↓
16	Phosphocholine	184.0	C5H14NO4P	6.559367	↓
17	PC(16:1(9Z)/2:0)	536.3	C26H50NO8P	6.133833	↓
18	Tanacetol B	314.2	C17H28O4	5.885183	↓
19	Estrone	315.1	C18H22O2	5.7404	↓
20	Assamsaponin B	659.2	C61H92O28	3.487067	↑
21	N-Acetyltyramine	180.1	C10H13NO2	2.83315	↓
22	Mammeigin	427.1	C25H24O5	0.494817	↓
23	PE-NMe(18:0/22:6(4Z, 7Z, 10Z, 13Z, 16Z, 19Z))	850.5	C46H80NO8P	7.21915	↑
24	Dityrosine	359.1	C18H20N2O6	3.810883	↓
25	PE(18:1(9Z)/0:0)	478.2	C23H46NO7P	6.485583	↓
26	LysoPC(20:0/0:0)	596.3	C28H58NO7P	6.8057	↓
27	PE-NMe2(18:0/20:4(5Z, 8Z, 11Z, 14Z))	794.5	C45H82NO8P	7.656367	↑
28	PE-NMe2(16:0/18:1(9Z))	766.5	C41H80NO8P	7.3342	↑
29	PE-NMe2(16:0/22:6(4Z, 7Z, 10Z, 13Z, 16Z, 19Z))	790.5	C45H78NO8P	7.21915	↓
30	LysoPC(O-18:0)	554.4	C26H56NO6P	6.78305	↓
31	Daphniphylline	526.4	C32H49NO5	6.569517	↓
32	4-Hydroxydebrisoquine	236.1	C10H13N3O	6.17285	↑
33	Bimatoprost	460.3	C25H37NO4	6.011733	↑
34	(S)-5, 7-Dihydroxy-6, 8-dimethylflavanone	329.1	C17H16O4	5.40835	↓
35	l-Glutamate	146.0	C5H9NO4	0.588583	↓

“↑” upregulated; “↓” downregulated, vs. Model.

Figure 5. Biomarker analysis and heatmap of identified marker metabolites. (A) Spearman correlation analysis among key efficacy indicators and marker metabolites. (B) Heatmap of selected marker metabolites among SXSH and SXSH-ABX group based on VIP > 1.

Data availability statement

The data generated for 16S rRNA gene profiling of bacteria are available in the NCBI repository with project accession number PRJNA900688.