Red Clover (Trifolium pratense) as a tool for helping with Blood sugar / diabetes support

Plant-derived exosomes (PDEs) are promising nanotherapeutics for improving chronic diseases, such as diabetes mellitus. Trifolium pratense (TP) is a flowering herb with potent antioxidant and antidiabetic properties. The present study aimed to explore the diabetic-healing effects of TP-derived exosomes (TPDEs) in streptozotocin (STZ)-induced diabetic rats. TPDEs were isolated using polyethylene glycol precipitation and serial centrifugation and characterised. STZ-induced diabetic rats were treated with TPDE doses (0, 100, 200, and 400 μg/kg) for 28 days. Biochemical factors (fasting blood sugar (FBS), insulin, C-peptide, total antioxidant capacity (TAC), and nitric oxide (NO)) were evaluated in serum samples. Also, the expression of PDX1, insulin, NGN3, and SIRT1 genes in pancreas tissues was assessed using real-time PCR. TPDE treatment decreased the serum levels of FBS and NO while increasing c-peptide, insulin, and TAC levels. It also significantly enhanced the expression of insulin, PDX1, NGN3, and SIRT1 genes. TPDEs at doses of 100 to 200 μg/kg showed the most significant antidiabetic effects. TPDEs significantly improved diabetes-induced alterations in serum insulin levels, antioxidant status, and pancreas-related gene expression. It can be considered a novel complementary treatment for diabetes.

Hot water extraction was applied to extract red clover (Trifolium pratense L.) polysaccharides (RCP) and the extraction conditions were optimized using the response surface methodology (RSM). An RCP yield of 12.72 ± 0.14% was achieved under the optimum extraction conditions: extracting time of 95 min, extracting temperature of 93 °C, and solvent-material ratio of 21 mL/g. A component named RCP-1.1 with the molecular weight of 7528.81 kDa was purified from RCP. RCP-1.1 was composed of glucose, galacturonic acid, arabinose, and galactose, with molar percentages of 52.54, 1.04, 16.31, and 30.11%, respectively. At the determination concentration of 10 mg/mL, the α-glucosidase inhibition ability of RCP-1.1 reached 86.72% of that of acarbose. The scavenging rates of RCP-1.1 (3.0 mg/mL) for DPPH and ABTS radicals reached 91.82% and 98.95% of that of ascorbic acid (3.0 mg/mL), respectively. Based on these results, RCP-1.1 possesses the potential to be used as a natural hypoglycemic agent or an antioxidant.

Natural plants have traditionally been used throughout the world for their anti-diabetic effects. The aim of the present study was to investigate the protective effect of hydroalcoholic extract of Trifolium pratens L. (T. pratense) on streptozotocin (STZ) cytotoxicity and insulin concentration from RIN-5F pancreatic β cell line. In this study, possible cytoprotective action of T. pratense extract (using pre-treatment, simultaneous, and post-treatment schedules) against STZ (30 mM) was evaluated using MTT assay. Apoptosis was quantified by fluorescent dye staining. Also, the effect of extract on insulin secretion in low and high glucose media was examined. Data were analyzed by one-way ANOVA test and P < 0.05 was considered significant. The viability of RIN-5F cells in 10, 20, 30, 40, and 60 μg/mL doses of T. pratense extract showed significant increases compared to control group (P < 0.001). STZ significantly reduced cell viability in a dose-dependent manner (P < 0.05). T. pratense extract in 20, 30, and 40 μg/mL doses had significant cytoprotective effect (P < 0.05) on cytotoxic action of STZ and this effect is greater in simultaneous treatment. STZ-mediated apoptotic death is reduced by extract. T. pratense extract treatment also, increased insulin concentration in cell culture medium. T. pratense had potent cytoprotective action, prevented apoptosis and increased insulin concentration in cell culture medium via the increase in pancreatic β cell number and/or insulin secretion. In addition, T. pratense enhanced viability of RIN-5F. Thus, T. pratense not only has anti-diabetic actions on β cells but also enhances their viability.