Influencia de bloqueadores de canais de calcio no processo de degeneração/regeneração muscular em camundongos ditroficos MDX / The influence iof calcium channel blockers in the process of muscular degeneration/regeneration in mdx mice

AUTOR(ES)
DATA DE PUBLICAÇÃO

2008

RESUMO

The lack of dystrophin in dystrophin-deficient fibers of mdx mice and in Duchenne muscular dystrophy leads to sarcolemmal breakdown and enhanced calcium influx followed by muscle degeneration.In this work, we examined whether the calcium channel blockers diltiazem and verapamil could protect dystrophic muscles from degeneration/regeneration. Mdx mice (n=32; 18 days old) received daily intraperitoneal injections of diltiazem (n=16; 72 mg/kg body weight) or verapamil (n=16; 25 mg/kg body weight) for 18 days, after which the sternomastoid, diaphragm, tibialis anterior and cardiac muscles were removed. Control mdx mice (n=16) were injected with saline. Both drugs significantly decreased the blood levels of creatine kinase (saline-treated mdx mice: 573±245 U/l, diltiazem-treated mice: 161±53* U/l and verapamil-treated mice: 217±57* U/l; mean+S.E.M., *p<0.05 vs. saline controls, Student´s t-test). The total calcium content, measured by plasma emission spectrometry, was 173-475% greater in mdx muscles compared to control C57Bl/10 muscles. Verapamil and diltiazem reduced the total calcium content only in diaphragm (diltiazem-treated mice: 229 mg calcium/kg vs. saline-treated mice: 295 mg calcium/kg; p=0.06, Student’s t-test) and cardiac muscle (diltiazem/verapamil-treated mice: 10 mg calcium/kg vs. saline-treated mice: 16 mg calcium/kg; p<0.05, Student’s t-test). Histological analysis showed that diltiazem significantly attenuated muscle degeneration only in diaphragm muscle (28% central nucleated fibers and 7% Evans blue-positive fibers in saline-treated mice vs. 12% central nucleated fibers and 1% Evans blue-positive fibers in diltiazem-treated mice; p<0.05, Student’s t-test). Immunoblots showed a significant increase in the levels of calsequestrin and β-dystroglycan in some diltiazem- and verapamil-treated muscles. Diltiazem was more effective than verapamil in protecting against muscle degeneration in mdx mice, especially in the more affected muscles. These results indicate that calcium channel blockers protect against muscle degeneration in the absence of dystrophin. They also suggest that these drugs could be useful therapeutic alternatives in the treatment of Duchenne muscular dystrophy. ABSTRACT The lack of dystrophin in dystrophin-deficient fibers of mdx mice and in Duchenne muscular dystrophy leads to sarcolemmal breakdown and enhanced calcium influx followed by muscle degeneration.In this work, we examined whether the calcium channel blockers diltiazem and verapamil could protect dystrophic muscles from degeneration/regeneration. Mdx mice (n=32; 18 days old) received daily intraperitoneal injections of diltiazem (n=16; 72 mg/kg body weight) or verapamil (n=16; 25 mg/kg body weight) for 18 days, after which the sternomastoid, diaphragm, tibialis anterior and cardiac muscles were removed. Control mdx mice (n=16) were injected with saline. Both drugs significantly decreased the blood levels of creatine kinase (saline-treated mdx mice: 573±245 U/l, diltiazem-treated mice: 161±53* U/l and verapamil-treated mice: 217±57* U/l; mean+S.E.M., *p<0.05 vs. saline controls, Student´s t-test). The total calcium content, measured by plasma emission spectrometry, was 173-475% greater in mdx muscles compared to control C57Bl/10 muscles. Verapamil and diltiazem reduced the total calcium content only in diaphragm (diltiazem-treated mice: 229 mg calcium/kg vs. saline-treated mice: 295 mg calcium/kg; p=0.06, Student’s t-test) and cardiac muscle (diltiazem/verapamil-treated mice: 10 mg calcium/kg vs. saline-treated mice: 16 mg calcium/kg; p<0.05, Student’s t-test). Histological analysis showed that diltiazem significantly attenuated muscle degeneration only in diaphragm muscle (28% central nucleated fibers and 7% Evans blue-positive fibers in saline-treated mice vs. 12% central nucleated fibers and 1% Evans blue-positive fibers in diltiazem-treated mice; p<0.05, Student’s t-test). Immunoblots showed a significant increase in the levels of calsequestrin and β-dystroglycan in some diltiazem- and verapamil-treated muscles. Diltiazem was more effective than verapamil in protecting against muscle degeneration in mdx mice, especially in the more affected muscles. These results indicate that calcium channel blockers protect against muscle degeneration in the absence of dystrophin. They also suggest that these drugs could be useful therapeutic alternatives in the treatment of Duchenne muscular dystrophy.

ASSUNTO(S)

mdx mice musculos verapamil muscle camundongo mdx degeneration calcium calcio diltiazem degeneração

ACESSO AO ARTIGO

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