Unveiling the Therapeutic Potential of Sarcopoterium spinosum Fruits: A Comprehensive Exploration from Chemical Characterization to Cellular Models of Health Challenges

The search for new bioactive compounds is of increasing interest, and it seems that despite new technical and scientific progress, most of the active ingredients have been known for centuries. Indeed, plants are an inexhaustible source of bioactive compounds that have been used since ancient times in both folk medicines and as preservatives of food. Medicinal plants have always been of interest for many kinds of industries because of their multiple applications for their antioxidant, antibacterial and cytoprotective properties. Plants, and plant-derived secondary metabolites may be applied in the management of a broad spectrum of metabolic dysfunction including obesity, fatty liver, cardiovascular disease. Metabolic diseases encompass a broad category of disorders characterized by disruptions of the body's metabolic homeostasis. These conditions impact on many metabolic pathways, such as glucose and lipid metabolism, with severe consequences for human health. Many phytochemicals being found in plants may exhibit diverse properties that can modulate metabolic pathways, enhance insulin sensitivity, and regulate lipid metabolism. Among them, Sarcopoterium spinosum has emerged as a noteworthy medicinal plant with the potential to address metabolic diseases. Indigenous to the Mediterranean region and the Middle East, this botanical species boasts a rich history in traditional medicine. Recent research has spotlighted its anti-diabetic properties, particularly in the context of its root extracts exhibiting the capacity to regulate blood glucose levels and enhance insulin sensitivity. A flourishing interest has emerged regarding its role as a natural remedy for metabolic disorders, contributing to the expanding array of medicinal plants with the potential to promote metabolic equilibrium and overall well-being. Notably, ongoing investigations have predominantly focused on the roots rather than the fruits of S. spinosum. In the context of this exploration, my PhD thesis aims to study different extracts derived from S. spinosum fruits, examining their bioactive compounds and their promising potential in ameliorating health and metabolic diseases. Three extracts were prepared from S. spinosum fruits, employing water, boiling water, and ethanol as extraction solvents. The ethanol extract, being distinguished by its superior radical scavenging potential, was selected for a detailed examination of the polyphenolic profile and the investigation of the potential beneficial effects. Two relevant cellular models were employed in this study: the rat hepatoma cell line (FaO) to mimic a cell model of hepatic steatosis, and the human endothelial cell line (HECV) to mimic a dysfunctional endothelium. The findings unveiled a rich profile of bioactive compounds for the ethanol extract, highlighting its abundance in ellagitannins, flavonoids, and terpenoids. Notably, this comprehensive chemical composition corresponded with the extract's demonstrated lipid-lowering, antioxidant, anti-inflammatory, and cytoprotective properties. In the final step of our investigation, our focus shifted to the design and develop a cellular model of insulin resistance utilizing the human hepatoma cell line HepG2. Employing hyperinsulinemia and fat accumulation as inducers, we successfully elicited the insulin resistance in vitro and this model will be tested using the S. spinosum extracts. In conclusion, the outcome of this PhD thesis shows that S. spinosum fruits could be a source of many bioactive compounds and the extracts represent a promising candidate to develop nutraceuticals or dietary supplements to treat/prevent obesity-related metabolic diseases

The Anti-Inflammatory Potential of an Ethanolic Extract from Sarcopoterium spinosum Fruits for Protection and/or Counteraction against Oxidative Stress in Dysfunctional Endothelial Cells

Plants and plant extracts are a relevant source of bioactive compounds widely employed as functional foods. In the Mediterranean area, the shrub Sarcopoterium spinosum is traditionally used as an herbal medicine for weight loss and a diabetes treatment. Inflammation is a protective mechanism involved in the development of many pathological conditions, including cardiovascular diseases. The present study aimed to investigate in vitro the antioxidant and cytoprotective properties of an ethanolic extract from S. spinosum fruits (SEE) in a cellular model of endothelium dysfunction. Corilagin and quercetin are two polyphenols abundant in SEE and were tested for comparison. The exposure of HECV cells for 24 h to 30 mu M hydrogen peroxide (H2O2) lead to an oxidative stress condition. When HECV cells were treated with 10 mu g/mL of SEE or single compounds after or before the oxidative insult, the results showed their ability to (i) decrease the reactive oxygen species (ROS) production quantified using fluorometric analysis and the lipid peroxidation measured with a spectrophotometric assay; (ii) rescue both the glutathione reduced to oxidized (GSH/GSSG) ratio and nitric oxide impair and the protein denaturation; and (iii) accelerate the wound repair measured using a T-scratch assay. Taken together, our findings indicate that the ethanolic extract from S. spinosum fruits could be a potential candidate for nutraceutical application

The Potential of Lamiaceae Herbs for Mitigation of Overweight, Obesity, and Fatty Liver: Studies and Perspectives

Numerous plants, plant extracts, and plant-derived compounds are being explored for their beneficial effects against overweight and liver diseases. Obesity is associated with the increased prevalence of non-alcoholic fatty liver disease (NAFLD), becoming the most common liver disease in Western countries. Obesity and NAFLD are closely associated with many other metabolic alternations such as insulin resistance, diabetes mellitus, and cardiovascular diseases. Many herbs of the Lamiaceae family are widely employed as food and spices in the Mediterranean area, but also in folk medicine, and their use for the management of metabolic disorders is well documented. Hereby, we summarized the scientific results of the medicinal and nutraceutical potential of plants from the Lamiaceae family for prevention and mitigation of overweight and fatty liver. The evidence indicates that Lamiaceae plants may be a cost-effective source of nutraceuticals and/or phytochemicals to be used in the management of metabolic-related conditions such as obesity and NAFLD. PubMed, Google Scholar, Scopus, and SciFinder were accessed to collect data on traditional medicinal plants, compounds derived from plants, their reported anti-obesity mechanisms, and therapeutic targets

Nuclear and chromatin rearrangement associate to epigenome and gene expression changes in a model of in vitro adipogenesis and hypertrophy

Hypertrophy of adipocytes represents the main cause of obesity. We investigated in vitro the changes associated with adipocyte differentiation and hypertrophy focusing on the nuclear morphometry and chromatin epigenetic remodelling. The 3 T3-L1 pre-adipocytes were firstly differentiated into mature adipocytes, then cultured with long-chain fatty acids to induce hypertrophy. Confocal and super-resolution stimulation emission depletion (STED) microscopy combined with ELISA assays allowed us to explore nuclear architecture, chromatin distribution and epigenetic modifications. In each condition, we quantified the triglyceride accumulation, the mRNA expression of adipogenesis and dysfunction markers, the release of five pro-inflammatory cytokines. Confocal microscopy revealed larger volume and less elongated shape of the nuclei in both mature and hypertrophic cells respect to pre-adipocytes, and a trend toward reduced chromatin compaction. Compared to mature adipocytes, the hypertrophic phenotype showed larger triglyceride content, increased PPAR & gamma; expression reduced IL-1a release, and up-regulation of a pool of genes markers for adipose tissue dysfunction. Moreover, a remodelling of both epigenome and chromatin organization was observed in hypertrophic adipocytes, with an increase in the average fluorescence of H3K9 acetylated domains in parallel with the increase in KAT2A expression, and a global hypomethylation of DNA. These findings making light on the nuclear changes during adipocyte differentiation and hypertrophy might help the strategies for treating obesity and metabolic complications

Polyphenol-enriched extracts of Sarcopoterium spinosum fruits for counteracting lipid accumulation and oxidative stress in an in vitro model of hepatic steatosis

Sarcopoterium spinosum (L.) Spach is a Rosaceae shrub employed in the folk medicine in the Eastern Mediterranean basin. The previous few studies have focused on the S. spinosum roots, while the fruits have been poorly investigated. The present study aims to assess the biological properties of S. spinosum fruits collected in Lebanon and subjected to ethanolic, water or boiling water extraction. The extracts were compared for the phenol and flavonoid contents, and for the in vitro radical scavenging ability. The ethanolic extract (SEE) was selected and characterized by high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS/MS) showing a phenolome rich in tannins (ellagitannins), flavonoids (quercetin derivatives), and triterpenes. The biological activity of SEE was tested on a cellular model of moderate steatosis consisting of lipid-loaded hepatic cells treated with increasing concentrations of SEE (1-25 mu g/mL), or with corilagin or quercetin as comparison. In steatotic hepatocytes the SEE was able (i) to ameliorate the hepatosteatosis; (ii) to counteract the excess ROS and lipid peroxidation; (iii) to restore the impaired catalase activity. The results indicate that the ethanolic extract from S. spinosum fruits is endowed with relevant antisteatotic and antioxidant activities and might find application as nutraceutical product

Influence of Simulated In Vitro Gastrointestinal Digestion on the Phenolic Profile, Antioxidant, and Biological Activity of Thymbra spicata L. Extracts

Plants or plant extracts are widely investigated for preventing/counteracting several chronic disorders. The oral route is the most common route for nutraceutical and drug administration. Currently, it is still unclear as to whether and how the pattern of phenolic compounds (PCs) found in the plants as well as their bioactivity could be modified during the gastrointestinal transit. Recent studies have revealed antioxidant and anti-steatotic properties of Thymbra spicata. Here, we investigated the possible loss of phytochemicals that occurs throughout the sequential steps of a simulated in vitro gastrointestinal (GI) digestion of aqueous and ethanolic extracts of aerial parts of T. spicata. Crude, digested, and dialyzed extracts were characterized in terms of their phenolic profile and biological activities. Total contents of carbohydrates, proteins, PCs, flavonoids, and hydroxycinnamic acids were quantified. The changes in the PC profile and in bioactive compounds upon the simulated GI digestion were monitored by HPLC–MS/MS analysis. The antioxidant activity was measured by different spectrophotometric assays, and the antiproliferative potential was assessed by using three representative human cancer cell lines. We observed that the simulated GI digestion reduced the phytochemical contents in both aqueous and ethanolic T. spicata extracts and modified the PC profile. However, T. spicata extracts improved their antioxidant potential after digestion, while a partial reduction in the antiproliferative activity was observed for the ethanolic extract. Therefore, our results could provide a scientific basis for the employment of T. spicata extract as valuable nutraceutical