Clinical evidence-guided network pharmacology analysis reveals a critical contribution of β1-adrenoreceptor upregulation to bradycardia alleviation by Shenxian-Shengmai

Figures

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  Fig. 1. Flow chart of article selection process. By the criteria shown on the right, finally 14 articles were screened out

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  Fig. 2. Assessment of the therapeutic effect of SXSM on patients with bradycardia. Analysis of heart rate shows that therapeutic effect percentage is 88.14% in SXSM group and 67.34% in the control group. Total 95%CI = 3.76. Heterogeneity I2 = 0%, Z = 7.47 (P < 0.00001) in total 957 cases. Risk of bias were evaluated for each research

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  Fig. 3. Risk of bias graph and funnel plot in meta-analysis. a Risk of bias graph. There were no high-risk literatures included in our study based on the seven assessment aspects of the Cochrane Collaboration Handbook standards. b Funnel plot. There was no small effect in the literature, indicating that the data obtained from different hospital subjects were basically the same, and the results were parallel comparable

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  Fig. 4. Effects of SXSM on acetylcholine-induced bradyarrhythmia in isolated rat heart. (A) representative ECG recordings of isolated and perfused rat hearts. Sample speed at 1k/s. Display of recorded results within 30 seconds. (a) untreated blank. (b)After acetylcholine perfusion. (c)After 1mg/ml SXSM perfusion. (B) Quantitation and statistical analysis of heart rate in each treatment group. There was no significant difference between the SXSM and blank groups. (C) Quantitation and statistical analysis of heart rate variability in each group. There was no significant difference between the SXSM and blank groups. Data are presented as mean ± SD (n =6). *P < 0.05

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  Fig. 5. Classification and prediction of component-related targets. Function and pathway prediction were carried out based on the target library established with chemical components in SXSM. (a) KEGG analysis of compounds targets in SXSM. Small red dots represent targets, colored nodes represent enriched pathways and functions. All prediction are based on gene annotation of KEGG online databases. (b) Top10 predicted signaling pathway. The order of top pathways are ranked by −log(p-value) and number of targets included

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  Fig. 6. Target network of bradycardia. (A) Disease related genes are displayed by a classification of location in body. Differenttypes are represented by different shapes. Most of the targets exist in the cell membrane, mainly ion channels and G protein-coupled receptors. (B) In the overview of adrenergic signaling pathway in cardiomyocytes, the ADR family is at a relatively upstream position (colored in red). Other identified targets by SXSM components are colored in pink or yellow

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  Fig. 7. Effects of SXSM on ADRB1 expression in cultured cardiomyocytes. (A) representative images of H9c2 cells with different treatments. (a) untreated control, (b) positive control (isoproterenol, 10 uM), (c) low dose of SXSM (0.5 mg/ml), (d) high dose of SXSM (1 mg/ml). Blue fluorescence represents nucleus stained by Hoechst, and yellow fluorescence represents ADRB1 protein which exists both on cell membrane and in cytoplasm. The expression level of ADRB1 was up-regulated, indicating that SXSM regulated β1-adrenergic signaling pathway activation. (B) Bar graph quantitation of ADRB1 levels in H9c2 cells with different treatments. n≥5, **means p<0.001, ***means p<0.0001 compared with control