The anti-lipopolysaccharide effect of glycine via fibroblastic cell culture from the embryonic calipash tissue of Chinese soft-shelled turtle Pelodiscus sinensis

Abstract

As a representative kind of soft-shelled turtles, Pelodiscus sinensis has soft apron termed calipash around its pliable carapace as the most distinctive trait, which is functionally important in turtle defense. It is of great significance to explore the physiological function of stress resistance and the possible regulation of anti-stress nutrition for the survival of aquaculture. To address this issue, a fibroblast cell line from the embryonic calipash tissue of P. sinensis (PSCF) was established from the perspective of ethics, repeatability, and convenience. Effect of glycine, the main amino acid component in calipash extracellular matrix, on the proliferation of PSCF treated with 160 μg/mL lipopolysaccharide (LPS) and the underlying mechanisms were also studied. The results showed that adding 100 μg/mL glycine significantly improved the fibroblast survival, increased the mRNA levels of collagen biosynthesis genes, prolyl hydroxylase and collagen degradation genes, and inhibited reactive oxygen species (ROS) production. It suggested that glycine shows positive effects on the cell proliferation of PSCF, and could regulate the oxidative stress and collagen metabolism. To our knowledge, this report is the first to describe cell cultures established at specific calipash tissue-forming stages during the embryonic development in P. sinensis and to reveal the anti-stress effect of glycine in turtles. The results will contribute to the application of functional calipash tissue, or its manipulation by nutrition additives, such as glycine, to prevent or ameliorate stress in turtle industry.