Myotoxicity induced by an acidic Asp-49 phospholipase A(2) isolated from Lachesis muta snake venom. Comparison with lysophosphatidylcholine.
In a previous report we showed that Lachesis muta crude venom displays potent indirect hemolytic activity and myotoxicity when injected into mice. Then, a phospholipase A(2) (PLA(2)) (LM-PLA(2)-I) responsible for these activities was isolated. More recently, a catalytically active isoenzyme (LM-PLA(2)-II) with molecular mass of 18 kDa and isoeletric point at pH 5.4 was isolated from the same snake venom. LM-PLA(2)-II inhibited ADP- and collagen-induced platelet aggregation as well as induced a potent paw edema reaction in rats. Here we show that LM-PLA(2)-II induced myotoxic effects both in vitro characterized by an increase on the rate of creatine kinase (CK) release from isolated mice extensor digitorum longus (EDL) muscles and in vivo by increasing plasma CK activity of injected mice. Histological analysis showed an intense damage in muscle cells injected with LM-PLA(2)-II. It was also shown that exogenous lysophosphatidylcholine (lyso-pc) behaved as a typical myotoxin damaging muscle cells, producing myonecrosis characterized by local infiltration of inflammatory cells similarly to that observed for LM-PLA(2)-II. Hemorrhage and lethal effects were not observed neither with LM-PLA(2)-II nor lyso-pc. As previously observed for other biological activities, pretreatment of LM-PLA(2)-II with p-bromophenacyl bromide (p-BPB) or acetic anhydride abolished all the enzyme's actions. The data confirms that biological activities displayed by LM-PLA(2)-II, including the myotoxic effects reported here, are all dependent on its enzymatic activity where the product formed (lyso-pc) may play an important function on such myotoxicity.
Fuly
André L
AL
Departamento de Bioquímica Médica, Universidade Federal do Rio de Janeiro, RJ, Rio de Janeiro, Brazil.
Calil-Elias
Sabrina
S
Martinez
Ana Maria B
AM
Melo
Paulo A
PA
Guimarães
Jorge A
JA
D000089:Acetic Anhydrides / Q000737:chemistry; D000098:Acetophenones / Q000737:chemistry; D000818:Animals; D003402:Creatine Kinase / Q000032:analysis / Q000097:blood / Q000378:metabolism; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology*; D008244:Lysophosphatidylcholines / Q000494:pharmacology / Q000633:toxicity*; D051379:Mice; D008854:Microscopy, Electron; D032342:Muscle Cells / Q000187:drug effects / Q000473:pathology / Q000648:ultrastructure; D009132:Muscles / Q000187:drug effects* / Q000473:pathology / Q000648:ultrastructure; D009336:Necrosis; D010741:Phospholipases A / Q000737:chemistry / Q000494:pharmacology / Q000633:toxicity*
D000089:Acetic Anhydrides; D000098:Acetophenones; D003435:Crotalid Venoms; C072644:Lachesis venom; D008244:Lysophosphatidylcholines; C031800:acetic anhydride; D003402:Creatine Kinase; D010741:Phospholipases A; C006821:4-bromophenacyl bromide