Impact of Using Supplemented Thylakoids Derived from Spinach for 12 Weeks of High-Intensity Functional Training on adipo-myokines in obese males

Asieh Abbassi Dalloii¹Maha Hoteit²Zahra Sadek²Mahboubeh Khak Rand¹Akbar Ramezani¹Zhaleh Pashaei³Mahsa Afshar⁴Kurt Escobar⁵Rashmi Supriya⁶Ayoub Saeidi^7*Hassane ZOUHAL⁸Ahmad Alkhatib⁹

• ¹Islamic Azad university, Amol, Iran
• ²Lebanese University, Beirut, Lebanon
• ³University of Tabriz, Tabriz, East Azerbaijan, Iran
• ⁴Islamic Azad University System, Tehran, Tehran, Iran
• ⁵California State University, Long Beach, Long Beach, California, United States
• ⁶Hong Kong Baptist University, Kowloon, Hong Kong, SAR China
• ⁷University of Kurdistan, Sanandaj, Kurdistan, Iran
• ⁸University of Rennes M2S (Laboratoire Mouvement, Sport, Santé), Rennes, France
• ⁹University of Taipei, Taipei, Taiwan

Objective: This randomized controlled study investigated the independent and combined effects of High-Intensity Functional Training (HIFT) and spinach-derived thylakoid supplementation on adipo-myokines, glycemic control, and lipid profiles in obese males. To compare the effects of HIFT alone, thylakoid supplementation (Thyl) alone, and their combination (HIFT+Thyl) on circulating adipokines (CTRP-2, CTRP-9, GDF-8, GDF-15), insulin resistance, and lipid profiles in obese adult males.Methods: A total of 68 participants who were obese with BMI: 32.6 ± 2.6 kg/m2 were randomly assigned to four groups (n = 17 in each group): thylakoid supplementation (Thyl), HIFT + Placebo High-Intensity Functional Training (HIFT), HIFT + thylakoid supplementation (HIFT+Thyl), and control+Placebo group (C). The training groups (HIFT and HIFT+Thyl) completed a 12week program of three 60-minute sessions per week. Participants in the Thyl and HIFT+Thyl groups dissolved and consumed 5 g/day of spinach extract high in thylakoids (or placebo) for 12 weeks. Baseline and post-intervention measurements included circulating C1Q/TNF or TGFβ related proteins (CTRP-2, CTRP-9, GDF-8, GDF-15), insulin resistance (HOMA-IR, plasma glucose, and insulin), lipid profile (HDL-C, LDL-C, triglycerides [TG], total cholesterol [TC]), and body composition (BMI, fat mass [FM], and fat-free mass [FFM]). Randomization was performed using a block randomization method with allocation concealment.Results: There were significant group x time interactions for all variables (all p < 0.001): CTRP-9 (η² = 0.6), CTRP-2 (η² = 0.7), GDF-8 (η² = 0.8), GDF-15 (η² = 0.4), BMI (η² = 0.45), FM (η² = 0.42), HDL-C (η² = 0.37), LDL-C (η² = 0.34), TC (η² = 0.46), TG (η² = 0.66), insulin (η² = 0.78), glucose (η² = 0.5), and HOMA-IR (η² = 0.7). Compared with baseline, all interventions (HIFT, Thyl, and HIFT+Thyl) significantly decreased adipokine levels (CTRP-9, CTRP-2, GDF-8, GDF-15), BMI, fat mass, LDL-C, TC, TG, insulin, glucose, and HOMA-IR, while increasing HDL-C (all p < 0.05). Post-hoc between-group comparisons showed that HIFT+Thyl resulted in significantly greater improvements in all adipo-myokines, lipid profile, glycemic and insulin control, and body fat compared to Thyl alone (all p < 0.05). HIFT and HIFT+Thyl showed comparable reductions in BMI, fat mass, and improvements in lipid profile and insulin sensitivity.