Gasparrini 2017 MiPschool Obergurgl
Event: MiPschool Obergurgl 2017
A common denominator in the pathogenesis of most chronic inflammatory diseases is the involvement of oxidative stress, related to ROS production by all aerobic organisms . ROS in particular represent the major source of cellular damage and the main site of their production is mitochondria: in normal conditions these reactive species are detoxified by specified enzymes; when these enzymes cannot transform ROS fast enough, oxidative damage occurs and accumulates in mitochondria, altering their metabolism and functionality . In recent years, different studies demonstrated that dietary antioxidants from plant foods could represent an efficient strategy to counteract this oxidative condition [2,3]. For this reason, we decided to evaluate the protective effects of Alba strawberry cultivar (100 µg/mL) and Manuka honey (3 mg/mL) extracts on inflammatory status induced by E. Coli LPS (1 µg/mL) on RAW 264.7 macrophages. In order to assess the effect of strawberry and Manuka treatment in presence or absence of LPS we measured (i) ROS intracellular production with the Tali® Image-Based cytometer and (ii) the oxygen consumption rate (OCR) related to mitochondria functionality, trough XF-24 Extracellular Flux Analyzer. After an OCR baseline measurement a profiling of mitochondrial function was performed by sequential injection of four compounds that affect bioenergetics, as follows: oligomycin, 2,4-Dinitrophenol (2,4 DNP) and antimycin A/rotenone. Moreover the maximal respiratory capacity value of each treatment was calculated with the following equation: OCR value(2, 4 DNP) – OCR value(antimycin A/rotenone). The protective effect of Alba and Manuka extract, at the different doses applied according to preliminary viability assays, on LPS-induced ROS production was demonstrated. Taking into account the OCR values, strawberry and Manuka honey treatments, significantly improved mitochondria functionality, also enhancing the increase in oxygen consumption and maximal respiratory capacity observed with LPS treatments (Fig. 1 and 2). Actually, there are no published studies that underline the effect of LPS-treatment on mitochondrial functionality of RAW macrophages, with particular attention to the OCR results. One possible explanation of our data could be related to the stimulatory effect exerted by the endotoxin LPS on RAW macrophages , which could produce an increase of oxygen consumption, despite the ROS results. However, further studies will be necessary to clarify this apparently contradictory aspect, also investigating the molecular pathway involved in mitochondria response after LPS and strawberry/Manuka honey treatments.
• Bioblast editor: Kandolf G
Labels: MiParea: Respiration, Comparative MiP;environmental MiP, Pharmacology;toxicology
Stress:Oxidative stress;RONS Organism: Mouse Tissue;cell: Macrophage-derived
Coupling state: LEAK Pathway: ROX
Event: B1, Poster
- Gasparrini M(1), Giampieri F(1), Forbes-Hernandez TY(2), Afrin S(1), Cianciosi D(1), Reboredo-Rodriguez P(3), Varela-Lopez A(4), Battino M(1,5)
- Dept Scienze Cliniche Specialistiche ed Odontostomatologiche, Univ Politecnica delle Marche
- Area Nutrición Salud, Univ Intern Iberoamericana
- Dept Quimica Analıtica Alimentaria, Univ Vigo
- Dept Physiol, Inst Nutrition Food Technol ‘‘José Mataix”, Univ Granada
- Centre Nutrition Health, Univ Europea Atlantico
- Spain.- firstname.lastname@example.org
Figure 1. OCR values of RAW macrophages subjected to the different treatments (ctrl, Alba, Manuka, LPS, Alba + LPS and Manuka + LPS). Mitochondria oxygen consumption was monitored with sequential injection of oligomycin, 2,4-DNP and antimycin A/rotenone at the indicated time points into each well, after baseline rate measurement. Data are expressed as mean values ± SD. Columns with different superscript letters are significantly different (P < 0.05).
Figure 2. Maximal respiratory capacity values of RAW macrophages subjected to the different treatments (ctrl, Alba, Manuka, LPS, Alba + LPS and Manuka + LPS). Data are expressed as mean values ± SD. Columns with different superscript letters are significantly different (P < 0.05).
- Giampieri F, Alvarez-Suarez JM, Battino M (2014) Strawberry and human health: effects beyond antioxidant activity. J Agric Food Chem 62:3867-76.
- Forbes-Hernandez TY, Gasparrini M, Afrin S, Mazzoni L, Reboredo P, Giampieri F (2016) A comparative study on cytotoxic effects of strawberry extract on different cellular models. J Berry Res 1-13.
- Alvarez-Suarez JM, Giampieri F, Cordero M, Gasparrini M, Forbes-Hernández TY, Mazzoni L, Afrin S, Beltrán-Ayala P, González-Paramás AM, Santos-Buelga C, Varela-Lopez A, Quiles JL, Battino M (2016) Activation of AMPK/Nrf2 signalling by Manuka honey protects human dermal fibroblasts against oxidative damage by improving antioxidant response and mitochondrial function promoting wound healing. J Funct Food 25:38–49.
- Zhang X, Xiong H, Liu L (2012) Effects of taraxasterol on inflammatory responses in lipopolysaccharide-induced RAW 264.7 macrophages. J Ethnopharmacol 141:206-11.