Citation: Stella, R.; Peggion, C.; Bergantin, C.; Biancotto, G.; Frosini, M.; Dreassi, E.; Marcolongo, P.; Aloisi, A.M.; Pessina, F. Serum Metabolomics and Proteomics to Study the Antihypertensive Effect of Protein Extracts from Tenebrio molitor. Nutrients 2022, 14, 3288. https:// doi.org/10.3390/nu14163288 Academic Editor: Kozo Nakamura Received: 8 July 2022 Accepted: 8 August 2022 Published: 11 August 2022 Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affil- iations. Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). nutrients Article Serum Metabolomics and Proteomics to Study the Antihypertensive Effect of Protein Extracts from Tenebrio molitor Roberto Stella 1, * ,† , Caterina Peggion 2,† , Caterina Bergantin 1 , Giancarlo Biancotto 1 , Maria Frosini 3 , Elena Dreassi 4 , Paola Marcolongo 5 , Anna Maria Aloisi 6 and Federica Pessina 5, * 1 Istituto Zooprofilattico Sperimentale delle Venezie, Department of Chemistry, viale dell’Università, 10-35020 Padova, Italy 2 Department of Biomedical Sciences, University of Padova, via Ugo Bassi 58/b, 35131 Padova, Italy 3 Department of Life Sciences, University of Siena, via A. Moro, 2-53100 Siena, Italy 4 Department of Biotechnology, Chemistry and Pharmacy, University of Siena, via A. Moro, 2-53100 Siena, Italy 5 Department of Molecular and Developmental Medicine, University of Siena, via A. Moro, 2-53100 Siena, Italy 6 Department of Medicine, Surgery and Neuroscience, University of Siena, via A. Moro, 2-53100 Siena, Italy * Correspondence: rstella@izsvenezie.it (R.S.); federica.pessina@unisi.it (F.P.) † These authors contributed equally to this work. Abstract: Hypertension is the leading risk factor for premature death worldwide and significantly contributes to the development of all major cardiovascular disease events. The management of high blood pressure includes lifestyle changes and treatment with antihypertensive drugs. Recently, it was demonstrated that a diet supplemented with Tenebrio molitor (TM) extracts is useful in the management of numerous pathologies, including hypertension. This study is aimed at unveiling the underlying mechanism and the molecular targets of intervention of TM dietary supplementation in hypertension treatment by means of proteomics and metabolomics techniques based on liquid chromatography coupled with high-resolution mass spectrometry. We demonstrate that serum proteome and metabolome of spontaneously hypertensive rats are severely altered with respect to their normotensive counterparts. Additionally, our results reveal that a diet enriched with TM extracts restores the expression of 15 metabolites and 17 proteins mainly involved in biological pathways associated with blood pressure maintenance, such as the renin–angiotensin and kallikrein–kinin systems, serin protease inhibitors, reactive oxygen scavenging, and lipid peroxidation. This study provides novel insights into the molecular pathways that may underlie the beneficial effects of TM, thus corroborating that TM could be proposed as a helpful functional food supplement in the treatment of hypertension. Keywords: functional food; multi-omics; blood pressure; SHR rats; insect protein supplementation 1. Introduction About a third of adults are affected by idiopathic hypertension, called essential hyper- tension, one of the most common contributors to mortality in the developed and developing world [1,2]. Hypertension represents an important issue in public health because of its increasing prevalence and its role in the development of several other diseases [3–5]. In fact, increased blood pressure is one of the most important comorbidities for the development of cardiovascular and cerebrovascular diseases, renal and heart failure, and stroke [6,7]. The mechanism of hypertension is complex as it arises from multiple genetic and environmental factors. Indeed, except for a few cases of patients with either renal or adrenal disease, there is no clear single identifiable cause in 95–97% of hypertension cases. Hypertension is probably caused by impairment of one of the numerous mechanisms involved in the main- tenance of normal blood pressure, which can differ among individuals. Such mechanisms may include, for example, (i) the renin–angiotensin–aldosterone system, consisting in a Nutrients 2022, 14, 3288. https://doi.org/10.3390/nu14163288 https://www.mdpi.com/journal/nutrients