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Diethylcarbamazine activity against Brugia malayi microfilariae is dependent on inducible nitric-oxide synthase and the cyclooxygenase pathway.

McGarry HF, Plant LD, Taylor MJ - Filaria J (2005)

Bottom Line: Pre-treatment of animals with dexamethasone or indomethacin reduced DEC's efficacy by almost 90% or 56%, respectively, supporting a role for the arachidonic acid and cyclooxygenase pathways in vivo.Furthermore, experiments showed that treatment with DEC results in a reduction in the amount of COX-1 protein in peritoneal exudate cells.These results confirm the important role of the arachidonic acid metabolic pathway in DEC's mechanism of action in vivo and show that in addition to its effects on the 5-lipoxygenase pathway, it targets the cyclooxygenase pathway and COX-1.

View Article: PubMed Central - HTML - PubMed

Affiliation: Filariasis Research Laboratory, Molecular and Biochemical Parasitology, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UK. hfcross@liverpool.ac.uk

ABSTRACT

Background: Diethylcarbamazine (DEC) has been used for many years in the treatment of human lymphatic filariasis. Its mode of action is not well understood, but it is known to interact with the arachidonic acid pathway. Here we have investigated the contribution of the nitric oxide and cyclooxygenase (COX) pathways to the activity of DEC against B. malayi microfilariae in mice.

Methods: B. malayi microfilariae were injected intravenously into mice and parasitaemia was measured 24 hours later. DEC was then administered to BALB/c mice with and without pre-treatment with indomethacin or dexamethasone and the parasitaemia monitored. To investigate a role for inducible nitric oxide in DEC's activity, DEC and ivermectin were administered to microfilaraemic iNOS-/- mice and their background strain (129/SV). Western blot analysis was used to determine any effect of DEC on the production of COX and inducible nitric-oxide synthase (iNOS) proteins.

Results: DEC administered alone to BALB/c mice resulted in a rapid and profound reduction in circulating microfilariae within five minutes of treatment. Microfilarial levels began to recover after 24 hours and returned to near pre-treatment levels two weeks later, suggesting that the sequestration of microfilariae occurs independently of parasite killing. Pre-treatment of animals with dexamethasone or indomethacin reduced DEC's efficacy by almost 90% or 56%, respectively, supporting a role for the arachidonic acid and cyclooxygenase pathways in vivo. Furthermore, experiments showed that treatment with DEC results in a reduction in the amount of COX-1 protein in peritoneal exudate cells. Additionally, in iNOS-/- mice infected with B. malayi microfilariae, DEC showed no activity, whereas the efficacy of another antifilarial drug, ivermectin, was unaffected.

Conclusion: These results confirm the important role of the arachidonic acid metabolic pathway in DEC's mechanism of action in vivo and show that in addition to its effects on the 5-lipoxygenase pathway, it targets the cyclooxygenase pathway and COX-1. Moreover, we show for the first time that inducible nitric oxide is essential for the rapid sequestration of microfilariae by DEC.

No MeSH data available.


Related in: MedlinePlus

DEC is ineffective against B. malayi microfilariae in the absence of iNOS. Efficacy of (a) a single, oral dose of DEC (100 mg/kg) or (b) a single, i.p. dose of ivermectin phosphate (1 mg/kg) in 129/SV and iNOS-/- mice infected with B. malayi microfilariae. Black symbols represent 129/SV mice, white symbols iNOS-/-. Squares indicate DEC administration, triangles ivermectin administration and circles untreated controls. Symbols represent mean results from at least three or four mice, except in the case of those treated with ivermectin (two animals) from two combined experiments which were representative of a further repeat. Significantly different microfilaraemias between 129/SV and iNOS-/- mice after DEC administration are denoted by * (P = 0.001) or ** (P = 0.000).
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Figure 3: DEC is ineffective against B. malayi microfilariae in the absence of iNOS. Efficacy of (a) a single, oral dose of DEC (100 mg/kg) or (b) a single, i.p. dose of ivermectin phosphate (1 mg/kg) in 129/SV and iNOS-/- mice infected with B. malayi microfilariae. Black symbols represent 129/SV mice, white symbols iNOS-/-. Squares indicate DEC administration, triangles ivermectin administration and circles untreated controls. Symbols represent mean results from at least three or four mice, except in the case of those treated with ivermectin (two animals) from two combined experiments which were representative of a further repeat. Significantly different microfilaraemias between 129/SV and iNOS-/- mice after DEC administration are denoted by * (P = 0.001) or ** (P = 0.000).

Mentions: DEC administration also rapidly reduced microfilaraemias in 129/SV mice but, in contrast, had no effect on microfilariae levels in iNOS-/- mice, in which microfilaraemia was maintained at pre-treatment levels for at least 2 hours (Fig. 3a), with no significant differences from untreated iNOS-/- controls (P > 0.887 for all time points). In contrast, ivermectin was effective in both 129/SV and iNOS-/- mice (Fig. 3b), although it had a slower onset of action than DEC. However, by 24 hours no microfilariae were detected in either strain of mouse given ivermectin.


Diethylcarbamazine activity against Brugia malayi microfilariae is dependent on inducible nitric-oxide synthase and the cyclooxygenase pathway.

McGarry HF, Plant LD, Taylor MJ - Filaria J (2005)

DEC is ineffective against B. malayi microfilariae in the absence of iNOS. Efficacy of (a) a single, oral dose of DEC (100 mg/kg) or (b) a single, i.p. dose of ivermectin phosphate (1 mg/kg) in 129/SV and iNOS-/- mice infected with B. malayi microfilariae. Black symbols represent 129/SV mice, white symbols iNOS-/-. Squares indicate DEC administration, triangles ivermectin administration and circles untreated controls. Symbols represent mean results from at least three or four mice, except in the case of those treated with ivermectin (two animals) from two combined experiments which were representative of a further repeat. Significantly different microfilaraemias between 129/SV and iNOS-/- mice after DEC administration are denoted by * (P = 0.001) or ** (P = 0.000).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC1173132&req=5

Figure 3: DEC is ineffective against B. malayi microfilariae in the absence of iNOS. Efficacy of (a) a single, oral dose of DEC (100 mg/kg) or (b) a single, i.p. dose of ivermectin phosphate (1 mg/kg) in 129/SV and iNOS-/- mice infected with B. malayi microfilariae. Black symbols represent 129/SV mice, white symbols iNOS-/-. Squares indicate DEC administration, triangles ivermectin administration and circles untreated controls. Symbols represent mean results from at least three or four mice, except in the case of those treated with ivermectin (two animals) from two combined experiments which were representative of a further repeat. Significantly different microfilaraemias between 129/SV and iNOS-/- mice after DEC administration are denoted by * (P = 0.001) or ** (P = 0.000).
Mentions: DEC administration also rapidly reduced microfilaraemias in 129/SV mice but, in contrast, had no effect on microfilariae levels in iNOS-/- mice, in which microfilaraemia was maintained at pre-treatment levels for at least 2 hours (Fig. 3a), with no significant differences from untreated iNOS-/- controls (P > 0.887 for all time points). In contrast, ivermectin was effective in both 129/SV and iNOS-/- mice (Fig. 3b), although it had a slower onset of action than DEC. However, by 24 hours no microfilariae were detected in either strain of mouse given ivermectin.

Bottom Line: Pre-treatment of animals with dexamethasone or indomethacin reduced DEC's efficacy by almost 90% or 56%, respectively, supporting a role for the arachidonic acid and cyclooxygenase pathways in vivo.Furthermore, experiments showed that treatment with DEC results in a reduction in the amount of COX-1 protein in peritoneal exudate cells.These results confirm the important role of the arachidonic acid metabolic pathway in DEC's mechanism of action in vivo and show that in addition to its effects on the 5-lipoxygenase pathway, it targets the cyclooxygenase pathway and COX-1.

View Article: PubMed Central - HTML - PubMed

Affiliation: Filariasis Research Laboratory, Molecular and Biochemical Parasitology, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UK. hfcross@liverpool.ac.uk

ABSTRACT

Background: Diethylcarbamazine (DEC) has been used for many years in the treatment of human lymphatic filariasis. Its mode of action is not well understood, but it is known to interact with the arachidonic acid pathway. Here we have investigated the contribution of the nitric oxide and cyclooxygenase (COX) pathways to the activity of DEC against B. malayi microfilariae in mice.

Methods: B. malayi microfilariae were injected intravenously into mice and parasitaemia was measured 24 hours later. DEC was then administered to BALB/c mice with and without pre-treatment with indomethacin or dexamethasone and the parasitaemia monitored. To investigate a role for inducible nitric oxide in DEC's activity, DEC and ivermectin were administered to microfilaraemic iNOS-/- mice and their background strain (129/SV). Western blot analysis was used to determine any effect of DEC on the production of COX and inducible nitric-oxide synthase (iNOS) proteins.

Results: DEC administered alone to BALB/c mice resulted in a rapid and profound reduction in circulating microfilariae within five minutes of treatment. Microfilarial levels began to recover after 24 hours and returned to near pre-treatment levels two weeks later, suggesting that the sequestration of microfilariae occurs independently of parasite killing. Pre-treatment of animals with dexamethasone or indomethacin reduced DEC's efficacy by almost 90% or 56%, respectively, supporting a role for the arachidonic acid and cyclooxygenase pathways in vivo. Furthermore, experiments showed that treatment with DEC results in a reduction in the amount of COX-1 protein in peritoneal exudate cells. Additionally, in iNOS-/- mice infected with B. malayi microfilariae, DEC showed no activity, whereas the efficacy of another antifilarial drug, ivermectin, was unaffected.

Conclusion: These results confirm the important role of the arachidonic acid metabolic pathway in DEC's mechanism of action in vivo and show that in addition to its effects on the 5-lipoxygenase pathway, it targets the cyclooxygenase pathway and COX-1. Moreover, we show for the first time that inducible nitric oxide is essential for the rapid sequestration of microfilariae by DEC.

No MeSH data available.


Related in: MedlinePlus