# Legumes as Functional Food for Cardiovascular Disease
## 5 Gene Network Analyses  
Based on this review, we performed a gene-to-gene network analysis using the Gene-MANIA plugin of the Cytoscape software platform [358; 359] and showed that 18 of the genes from all of these legume studies are functionally interrelated (Figure 4). Meanwhile, pathway and gene ontology (GO) terms enrichment using the Reactome functional interaction program [360] revealed that most of these genes are involved in lipid transport, fatty acid, and TAG metabolic processes and in the regulation of these processes (Table 3), suggesting that these pathways may be key and/or target processes to understand the  
Figure 3: Hydrophobicity analysis of the 22 peptides discussed in this manuscript. **(a)**. Boxplot of the Grand average of hydropathicity index (GRAVY) values of these peptides calculated through the Protein GRAVY website ([[https://www.bioinformatics.org/sms2/protein_gravity.html](https://www.bioinformatics.org/sms2/protein_gravity.html)]([https://www.bioinformatics.org/sms2/protein_gravity.html](https://www.bioinformatics.org/sms2/protein_gravity.html)) accessed on 11 February 2021) [355] and visualized using RStudio version 1.1.456 (Rstudio, Boston, MA, USA). GRAV scores greater than 0 are considered hydrophobic while scores less than 0 are considered hydrophilic [354; 356] as demarcated by the brown, dashed, horizontal line. **(b)** Proportion of the 22 peptides classified based on their hydrophobicity GRAVY scores. Figure 2: Histogram of the frequency of the lengths of various peptides (\(n=22\)) discussed in this manuscript. The mean number of amino acid residues is indicated as a brown-colored line on the \(x\)-axis. mechanism of action of these legume proteins. Notably, genes such as _CPTIA_, _SREBF2_, _PPARG_, and _LDLR_ are shown to be commonly involved in pathways related to lipid and fatty acid metabolism. These may serve as target genes and proteins for further studies to elucidate how legume and their proteins affect these pathways as well as how these genes regulate other genes and proteins such as those involved in cardiovascular health. The Mechanism of Action (MOA) of Legumes Proteins and Peptides Associated with Cardiovascular Health  
Based on the comprehensive literature search [361; 362; 363], the known genes, proteins, enzymes and corresponding cellular localization have been integrated to depict the highly possible mechanisms of action of isolated legume proteins and/or peptides in relation to their cardiovascular benefits (Figure 5). This figure illustrates how the induction of these protein and peptide isolates affects the cellular physiology and metabolism in the hepatic and blood serum systems that can lead to improved cardiovascular health. Two major pathways are emphasized in these mechanisms of action: (a) cholesterol and fatty acid degradation and (b) the inhibition of angiotensin converting enzyme (ACE). The first pathway showed a series of mechanisms with the ultimate goal of lowering lipid accumulation in the system. In order to decrease the lipid levels, legume protein isolates target cholesterol and fatty acid metabolism. LDL receptor genes are observed to be upregulated in order to produce more LDR receptor proteins that can receive and engulf LDL and VLDL for subsequent lysosomal degradation. Essential nuclear transcription regulators like SREBPP1c and SREBPP-2 play critical roles in lowering cholesterol and fatty acid synthesis. Aside from lowering lipid and cholesterol levels, such processes subsequently lower fat accumulation and an increased fat excretion. The second major effect of legume isolates is their ability to regulate hypertension. From the available literature, results suggest that Angiotensin I and II are regulated by blocking the functions of ACE through an unknown mechanism. This observation leads to more relaxed blood vessels, lesser vasoconstriction, lowered blood pressure, and ultimately contributing to improved cardiovascular health. Figure 4: Functional network of genes associated with cardiovascular disease and legumes proteins.