Impact of Lycium barbarum polysaccharide on apoptosis in Mycoplasma-infected splenic lymphocytes
Purpose: To evaluate the effect of Lycium barbarum polysaccharide (LBP) on apoptosis in Mycoplasma-infected splenic lymphocytes (SLs), and the underlying mechanisms.
Methods: SLs isolated from C57BL/6J mice were infected with Mycoplasma. The infected SLs were administered at different concentrations of LBP for 4 h, and the proportions of apoptotic cells and levels of relative reactive oxygen species (ROS) were determined by flow cytometry. The expressions of proapoptotic genes and endogenous antioxidant enzymes were investigated by real-time polymerase chain reaction (RT-PCR) and Western blotting.
Results: LBP treatment produced dose-dependent reductions in apoptotic ratio and intracellular ROS levels of SLs (p < 0.05). In addition, the expressions of pro-apoptotic genes were decreased by LBP treatment with respect to mRNA and protein levels (p < 0.05). In contrast, mRNA and protein levels of anti-apoptotic factor Bcl-2 were significantly increased in a dose-dependent manner (p < 0.05). Furthermore, RT-PCR and Western blot results demonstrated that the expression levels of mRNA and proteins in Nrf2, HO-1 and NQO1 were up-regulated by Mycoplasma infection (p < 0.01), and further increased by LBP treatment (p < 0.05).
Conclusion: LBP exerts a hyperactive antioxidant response encoded by Nrf2 to protect SLs from apoptosis induced by ROS-related oxidative damage after Mycoplasma infection. These results suggest that LBP may serve as a beneficial and dietary anti-Mycoplasma and anti-apoptotic agent.
Keywords: Lycium barbarum polysaccharide, Splenic lymphocytes, ROS, Caspase-3, Bax, Nrf2