lithium-chloride and Lupus-Erythematosus--Systemic

The abnormal activation of the AKT/GSK3β signal pathway in lymphocytes from systemic lupus erythematosus (SLE) patients plays an important role in the pathogenesis of the disease. Recently Hydrogen sulfide (H2S) has been recognized as a crucial gaseous signaling molecule, involved in regulation of cell proliferation. However, the role of H2S in regulating the abnormal activation of lymphocytes from SLE patients has not been established. This study was conducted to investigate the effect of H2S on lymphocytes and to explore the mechanisms involved.. The lymphocytes were isolated from SLE patients with or without renal disease and healthy controls. The cells were treated as indicated in each experiment. Cell viability was analyzed by CCK-8. Cell cycle distribution was determined by flow cytometry. Western blot was used to detect the expression of phosphorylated AKT (ser473), GSK3β (ser9) and CDK2, p27(Kip1) and p21(WAF1/CIP1).. Our findings showed that proliferation of lymphocytes was stimulated following treatment with NaHS (a H2S donor) at low NaHS concentrations (<1mM) but inhibited at high NaHS concentrations (>2mM). Similar results were observed using GYY4137, which is a slow-releasing H2S donor. Pretreatment of lymphocytes from SLE patients with NaHS at high concentrations prior to exposure to phytohemagglutinin (PHA) significantly attenuated proliferation, evidenced by decrease in cell viability and S phase distribution of cell cycle. Pretreatment with NaHS decreased PHA-induced expression of CDK2, phosphorylation levels of AKT (ser473) and GSK3β (ser9) and increased the expression of p27(Kip1) and p21(WAF1/CIP1). Moreover, pretreatment with NaHS blunted the stimulation of SLE lymphocyte proliferation by GSK3β inhibitor lithium chloride.. These results demonstrate that H2S inhibits the abnormal activation of lymphocytes from SLE patients throuqh the AKT/GSK3β signal pathway.

Topics: Cell Proliferation; Cell Survival; Cells, Cultured; Cyclin-Dependent Kinase 2; Cyclin-Dependent Kinase Inhibitor p21; Cyclin-Dependent Kinase Inhibitor p27; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Humans; Hydrogen Sulfide; Lithium Chloride; Lupus Erythematosus, Systemic; Lymphocytes; Morpholines; Organothiophosphorus Compounds; Phosphorylation; Phytohemagglutinins; Proto-Oncogene Proteins c-akt; S Phase Cell Cycle Checkpoints; Signal Transduction

Daily administration of 4 mg 6LiCl to groups (15 mice/group) of female NZB/W mice starting at 8 weeks of age led to long-term survival (44 weeks of age) of 73% of the mice when injections were performed between 08:00 and 10:00 h and 67% of mice when injections were performed between 17:00 and 19:00 h. Untreated controls were dead by 34 weeks of age and the differences between untreated and treated groups was significant (P < or = 10(-4)). In contrast daily administration of melatonin (100 micrograms/mouse) did not significantly enhance survival when injections were performed between 17:00 and 19:00 h but did enhance survival when given between 08:00 and 10:00 h (P < or = 10(-3)). Differences between the two melatonin groups was also significant (P < or = 0.05). Mice treated with Li plus melatonin exhibited survival curves identical to mice treated with Li alone. Therefore, the Li effect was dominant and survival was not altered by melatonin. Cessation of treatment in long-term survivors at 44 weeks of age led to the rapid death of 80% of the mice previously treated between 17:00 and 19:00 h (Li, Li + melatonin). In contrast, only 40% of the long-term survivors in the other groups had died by 66 weeks of age (22 weeks post-treatment). Thus the p.m. groups were less protected from disease reactivation than were the a.m. groups. Neither Li, melatonin, nor Li+melatonin influenced anti-gp70 or anti-ssDNA levels in serum, but Li treatment maintained renal function as determined by proteinuria scores. These findings indicate that the effectiveness of Li is probably not related to melatonin metabolism and immunomodulating influences, but the influence of other neuroendocrine variables cannot be eliminated.

Topics: Adjuvants, Immunologic; Aging; Animals; Antibodies, Antinuclear; Disease Progression; DNA, Single-Stranded; Enzyme-Linked Immunosorbent Assay; Female; Indicators and Reagents; Kidney; Lithium Chloride; Lupus Erythematosus, Systemic; Melatonin; Mice; Mice, Inbred Strains; Proteinuria; Time Factors

Previous investigations have indicated that initiation of LiCl treatment (4 mg/day) of female NZB/W mice at 10 weeks of age led to enhanced survival of 50% of the mice to > 11 months of age (Lithium, 3, 61-67, 1992). The present results indicate that this enhancement of survival is dose dependent in that 2 mg 7LiCl/day was less effective than 4 mg 7LiCl/day. Daily treatment of groups of mice with 2 or 4 mg LiCl per day starting at 8 weeks of age led to the survival of 40% and 70%, respectively, of the mice at 40 weeks of age, a time when only 10% of the untreated mice remained alive. Initiation of treatment with 2 mg 7LiCl/day after the disease process was evident (24 weeks of age) led to diminished effectiveness. Parallel experiments with mice treated with 4 mg 7LiCl/day revealed 60% long-term survivors in the early treatment groups and 33% in the delayed treatment groups. Cessation of treatment in both groups led to some additional deaths, but 20-27% of the mice remained alive at 60 weeks of age, even though animals had detectable levels of anti-ssDNA antibodies in their serum. Additional experiments with 2 and 4 mg 7LiCl/day in NZB/W mice pretreated with C. parvum-PER, a treatment previously shown to enhance survival (Int. J. Immunopharmac., 14, 35-41, 1992), indicated that the effect of the two modalities was not additive. The results presented indicate that treatment of NZB/W mice with LiCl leads to very effective prolongation of survival by unique mechanisms, possibly involving alteration of the effector phase of autoimmune damage to the kidney or the susceptibility of kidney elements to immune-mediated damage leading to renal failure.

Topics: Animals; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Kidney; Lithium Chloride; Lupus Erythematosus, Systemic; Mice; Mice, Inbred NZB; Propionibacterium acnes; Survival Rate; Time Factors