| Sertoli cells(SC)are epithelial cells that initiate sexual function development in male animals and maintain spermatogenesis after sexual maturation.It can directly contact with spermatogenic cells and provide nutrition for them.It can also secrete a variety of cytokines to maintain the process of spermatogenesis,and play an important role in maintaining the spermatogenic environment of testis,promoting the proliferation and differentiation of spermatogenic cells and ensuring the spermatogenic ability of males.Therefore,SC can determine the quality of sperm,and its proliferation and biological function have a significant impact on porcine fertility.Fluoride is a halogen element widely existing in underground water.Studies have shown that high dose of fluoride can not only cause damage to various types of cells,but also penetrate the blood-testosterone barrier of male animals,thus causing the decline of their reproductive ability.It mainly leads to impaired cell function by stimulating oxidative stress and eventually leads to apoptosis.Glycine is a recognized free radical scavenger and belongs to the antioxidant system in cells.Studies have shown that glycine has a variety of physiological functions,including anti-oxidation,anti-apoptosis,improve immunity and so that plays an important role in maintaining the life activities of the body.In order to explore the protective mechanism of glycine against excessive fluoride exposure induced SC injury in porcine testis,we treated porcine testis SC separately or simultaneously with glycine and sodium fluoride to detect the changes in its physiological state and function,and tried to elucidate some of the molecular mechanisms in the process of cellular changes.The specific content is as follows:In this study,the effects of different concentrations of sodium fluoride on SC activity of porcine testis and the protective ability of different concentrations of glycine on SC activity were firstly tested.Then the effects of glycine on SC cell cycle,proliferation and migration induced by sodium fluoride were investigated.It was found that 800 μm sodium fluoride could significantly inhibit the activity of SC(P <0.001)and the proliferation ability of cells in the testis of porcine(P < 0.05).The migration ability gradually decreased with the increase of time.When the time reached 120 h,the migration ability was significantly decreased(P < 0.001).The 800 μm sodium fluoride also resulted in a very significant block of SC in the G1 phase(P < 0.01),which resulted in a very significant decrease in the number of cells in the S phase(P < 0.001).Meanwhile,the addition of 2 m M glycine could effectively alleviate the adverse effect of sodium fluoride on SC proliferation and restore the proliferation ability of cells to the normal level.Subsequently,the levels of intracellular oxidative stress,mitochondrial function,DNA damage and apoptosis during the treatment were measured in order to understand the physiological processes and molecular mechanisms involved in glycine protection against excess sodium fluoride exposure induced SC damage in porcine testis.The results showed that glycine could significantly inhibit the production of ROS in SC induced by sodium fluoride(P < 0.0001)and play a role in supplementing GSH to a certain extent.In addition,glycine can significantly improve mitochondrial dysfunction and restore mitochondrial membrane potential(P < 0.0001)and promote the production of ATP(P < 0.05),thereby inhibiting the accumulation of DNA damage(P < 0.01)and decreased the incidence of apoptosis(P < 0.001).Glycine supplementation also significantly down-regulated the expression of aging markers such as P53,P21,HMGA2 and p16INK4 A genes in high fluorine-induced testis SC(P < 0.05)and alleviate the aging process of SC in porcine testis induced by sodium fluoride.In this study,it was found that glycine had a significant protective effect on the proliferation and damage of porcine testicular SC induced by sodium fluoride.This mechanism mainly played a role by inhibiting mitochondrial apoptosis pathway,and prevented cell DNA damage and even apoptosis on the premise of protecting mitochondrial function by inhibiting oxidative stress.These results provide a theoretical basis for promoting the proliferation of pig testicular SC,alleviating the damage and apoptosis of SC under oxidative stress stimulation,and provide a new insight for understanding the role of glycine in SC protection. |
