Lymphocyte apoptosis induced by sulfur mustard in rat spleen / 解放军医学杂志

ABSTRACT

Objective To investigate the apoptotic effect induced by sulfur mustard on the lymphocytes of the rat spleen,and define the role of caspase-3 during the process.Methods Sulfur mustard was given by intraperitoneal injection in a dose of 3.5mg/kg.The animals were anesthetized and the spleens were harvested at different timepoints after intoxication.The histopathogy of spleen was studied with hematoxylin-eosin staining.Caspase-3 mRNA was detected with RT-PCR.Protein expression of caspase-3 was assayed with Western blotting,lymphocytes of rat spleen were isolated and cultured in vitro and they were challenged with sulfur mustard(100?mol/L).The effect of Ac-DEVD-CHO(a specific inhibitor of caspase-3)on sulfur mustard-induced apoptosis of the lymphocytes cultured in vitro was evaluated.Fluorescent probe labeled with Rhodamine 123 was used to study mitochondrial potential.Results The histology of rat spleen was affected after sulfur mustard intoxication,as evidenced by apoptosis of a part of lymphocytes.Protein and mRNA expressions of caspase-3 were increased significantly in the spleens of intoxicated rats as compared with that in control group.DNA ladder and 'sub-G1' peak of lymphocytes which were treated with sulfur mustard in vitro were partially improved by Ac-DEVD-CHO(the specific inhibitor of caspase-3).In addition,mitochondrial potential decreased in a time-dependent manner in the lymphocytes intoxicated by sulfur mustard in vitro compared with that in the control group.Conclusions The spleen is injured in the rat which is intoxicated by sulfur mustard.Lymphocyte apoptosis is one of the mechanisms splenic injury.Caspase-3 may be involved in the process of lymphocyte apoptosis induced by sulfur mustard.