Dias 2017 MiPschool Obergurgl
Event: MiPschool Obergurgl 2017
Sertoli cells (SCs) play a key role in spermatogenesis support. Their high metabolic rates and mitochondrial activity are associated with a high production of reactive oxygen intermediates (ROIs) . Any disruptions in SCs function may lead to an uncontrolled ROIs production, compromising male fertility. The tea components epigallocatechin-3-gallate (EGCG) and L-theanine (L-The) have shown a protective role against toxicity in certain cell types [2,3]. Thus, we evaluated the effects of EGCG and L-the on the metabolism, mitochondrial functionality and oxidative profile of cultured human Sertoli cells (hSCs).
Firstly, hSCs were cultured in the absence or presence of 50 μM EGCG or L-The. After 24h of exposure, hSCs proliferation was evaluated by the sulforhodamine B assay. Glucose consumption and lactate production were measured in hSCs extracellular media by 1H-NMR. Then, the protein levels of mitochondrial complexes I-V were evaluated by Western blot, while the mitochondrial membrane potential was evaluated using the JC-1 probe. We also performed an O2 consumption rate assay using the MitoXpress kit. Oxidative damage levels to proteins and lipids were evaluated by the immunoblot technique using specific antibodies.
Cell proliferation decreased in hSCs exposed to EGCG (Figure 1) but increased in those exposed to L-the (Figure 2). Both groups of hSCs cultured with EGCG or L-the demonstrated a higher glucose uptake, tough lactate production was maintained. Interestingly, while EGCG induced a decrease in mitochondrial membrane potential, L-the led to a mitochondrial hyperpolarization. However, O2 consumption rate was not affected by EGCG neither by L-the. EGCG could also decrease hSCs lipid peroxidation.
In conclusion, EGCG and L-the triggered opposite effects in hSCs function, but probably with similar outcomes. There seems to be a relation with hSCs proliferation and mitochondrial membrane potential since both decreased (EGCG) or increased (L-the). Further work is needed to understand if these scenarios can compromise ATP production rates, although lactate production was maintained and ROIs production controlled, which could be beneficial for the nutritional support of spermatogenesis, particularly under unfavorable conditions.
• Bioblast editor: Kandolf G
Labels: MiParea: Exercise physiology;nutrition;life style, Pharmacology;toxicology
Organism: Human Tissue;cell: Genital
Enzyme: Complex I, Complex II;succinate dehydrogenase, Complex III, Complex IV;cytochrome c oxidase, Complex V;ATP synthase
Event: D2, Oral
- Dias TR(1,2,3), Bernardino RL(2), Alves MG(2), Silva J(4), Barros A(4,5,6), Sousa M(2,4), Casal S(3), Silva BM(1), Oliveira PF(2,5,6)
- Centro Investigação Ciências Saúde (CICS-UBI), Univ Beira Interior, Covilhã
- Dept Microscopy, Lab Cell Biol, Inst Biomedical Sciences Abel Salazar & Unit Multidisciplinary Research Biomedicine, Univ Porto
- LAQV/REQUIMTE — Lab Bromatol Hydrol, Fac Pharmacy, Univ Porto
- Centre Reproductive Genetics Prof. Alberto Barros, Porto
- Dept Genetics, Fac Medicine, Univ Porto
- i3S- Inst Investigação Inovação Saúde, Univ do Porto
- Portugal. – [email protected]
Figure 1. Effect of epigallocatechin-3-gallate in human Sertoli cells proliferation (left) and mitochondrial membrane potential (right). Cell proliferation data are presented as percentage, where control value was set at 100%, while that of mitochondrial membrane potential are presented by the JC-1 ratio as fold variation to the control. All results are expressed as mean ± SEM (n=6 for each condition). Significantly different results (P<0.05) are indicated as: a – relative to control.
Figure 2. Effect of L-theanine in human Sertoli cells proliferation (left) and mitochondrial membrane potential (right). Cell proliferation data are presented as percentage, where control value was set at 100%, while that of mitochondrial membrane potential are presented by the JC-1 ratio as fold variation to the control. All results are expressed as mean ± SEM (n=6 for each condition). Significantly different results (P<0.05) are indicated as: a – relative to control.
References and support
- Dias TR, Martins AD, Reis VP, Socorro S, Silva BM, Alves MG, Oliveira PF (2013) Glucose transport and metabolism in sertoli cell: relevance for male fertility. Current Chem Biol 7:282-93.
- Rizvi SI, Zaid MA, Anis R, Mishra N (2005) Protective role of tea catechins against oxidation‐induced damage of type 2 diabetic erythrocytes. Clinic Experim Pharmacol Physiol 32:70-5.
- Li G, Ye Y, Kang J, Yao X, Zhang Y, Jiang W, Gao M, Dai Y, Xin Y, Wang Q (2012) l-Theanine prevents alcoholic liver injury through enhancing the antioxidant capability of hepatocytes. Food Chem Toxic 50:363-72.
- Selected mentor: Prof. Dr. Carlos Palmeira