Analysis of immunomodulating nitric oxide, iNOS and cytokines mRNA in mouse macrophages induced by microcystin-LR.
Microcystins (MCs) are the toxic molecules produced by common cyanobacterium in freshwater blooms. Their toxicities have brought severe health issues to livestock and human being. Microcystin-LR (MC-LR) is one of the most toxic MCs. This paper presents the profile of the immunomodulation of MC-LR to BALB/c mice peritoneal macrophages. Macrophages were stimulated with 100microg/l lipopolysaccharide (LPS) and MC-LR at dose of 1, 10, 100, 1000nmol/l, respectively, for 24h. Nitric oxide (NO) production in cell culture supernatants was quantified by using Griess reagent method. Total RNA was extracted from incubated macrophages then the mRNA abundance of induced nitric oxide synthase (iNOS), IL-1beta, TNF-alpha, GM-CSF, IFN-gamma was monitored by using reverse-transcriptional polymerase chain reaction (RT-PCR). The results demonstrated that NO production, mRNA levels of iNOS, IL-1beta, TNF-alpha were down regulated by MC-LR dose-dependently and mRNA levels of GM-CSF and IFN-gamma were also decreased but in dose-independent manner. Our results illustrated the involvement of NO production, iNOS and some cytokines in mice immune system in microcystin shock.
Department of Biochemistry, State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, PR China.
Ting
T
Chen
Xiangyi
X
Zhao
Yang
Y
Liu
Qian
Q
Shi
Zichun
Z
Hua
Pingping
P
Shen
D000276:Adjuvants, Immunologic / Q000633:toxicity*; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000737:chemistry; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects; D008070:Lipopolysaccharides / Q000494:pharmacology; D017737:Macrophages, Peritoneal / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D008387:Marine Toxins / Q000633:toxicity*; D051379:Mice; D008807:Mice, Inbred BALB C; D052998:Microcystins; D008934:Mitogens / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000235:genetics / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II; D010456:Peptides, Cyclic / Q000633:toxicity*; D012333:RNA, Messenger / Q000378:metabolism
D000276:Adjuvants, Immunologic; D017077:Culture Media, Conditioned; D016207:Cytokines; D008070:Lipopolysaccharides; D008387:Marine Toxins; D052998:Microcystins; D008934:Mitogens; D010456:Peptides, Cyclic; D012333:RNA, Messenger; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; C057862:cyanoginosin LR