Proteins from latex of Calotropis procera prevent septic shock due to lethal infection by Salmonella enterica serovar Typhimurium.
OBJECTIVE
The latex of Calotropis procera has been used in traditional medicine to treat different inflammatory diseases. The anti-inflammatory activity of latex proteins (LP) has been well documented using different inflammatory models. In this work the anti-inflammatory protein fraction was evaluated in a true inflammatory process by inducing a lethal experimental infection in the murine model caused by Salmonella enterica Subsp. enterica serovar Typhimurium.


METHODS
Experimental Swiss mice were given 0.2 ml of LP (30 or 60 mg/kg) by the intraperitoneal route 24 h before or after lethal challenge (0.2 ml) containing 10(6) CFU/ml of Salmonella Typhimurium using the same route of administration.


RESULTS
All the control animals succumbed to infection within 6 days. When given before bacterial inoculums LP prevented the death of mice, which remained in observation until day 28. Even, LP-treated animals exhibited only discrete signs of infection which disappeared latter. LP fraction was also protective when given orally or by subcutaneous route. Histopathological examination revealed that necrosis and inflammatory infiltrates were similar in both the experimental and control groups on days 1 and 5 after infection. LP activity did not clear Salmonella Typhimurium, which was still present in the spleen at approximately 10(4) cells/g of organ 28 days after challenge. However, no bacteria were detected in the liver at this stage. LP did not inhibit bacterial growth in culture medium at all. In the early stages of infection bacteria population was similar in organs and in the peritoneal fluid but drastically reduced in blood. Titration of TNF-alpha in serum revealed no differences between experimental and control groups on days 1 and 5 days after infection while IL-12 was only discretely diminished in serum of experimental animals on day 5. Moreover, cultured macrophages treated with LP and stimulated by LPS released significantly less IL-1beta.


CONCLUSIONS
LP-treated mice did not succumb to septic shock when submitted to a lethal infection. LP did not exhibit in vitro bactericidal activity. It is thought that protection of LP-treated mice against Salmonella Typhimurium possibly involves down-regulation of pro-inflammatory cytokines (other than TNF-alpha). LP inhibited IL-1beta release in cultured macrophages and discretely reduced IL-12 in serum of animals given LP. Results reported here support the folk use of latex to treat skin infections by topic application.
Lima-Filho
José V
JV
Departamento de Biologia, Universidade Federal Rural de Pernambuco, Campus Dois Irmãos, Recife, PE, Brazil. jvitor@db.ufrpe.br
Patriota
Joyce M
JM
Silva
Ayrles F B
AF
Filho
Nicodemos T
NT
Oliveira
Raquel S B
RS
Alencar
Nylane M N
NM
Ramos
Márcio V
MV
D000818:Animals; D000900:Anti-Bacterial Agents / Q000302:isolation & purification / Q000494:pharmacology / Q000627:therapeutic use*; D031107:Calotropis / Q000737:chemistry*; D002478:Cells, Cultured; D015169:Colony Count, Microbial; D016207:Cytokines / Q000276:immunology; D004195:Disease Models, Animal; D007840:Latex / Q000737:chemistry; D008099:Liver / Q000187:drug effects / Q000473:pathology; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000276:immunology; D008297:Male; D051379:Mice; D010940:Plant Proteins / Q000302:isolation & purification / Q000494:pharmacology / Q000627:therapeutic use*; D012480:Salmonella Infections / Q000276:immunology / Q000382:microbiology / Q000473:pathology / Q000517:prevention & control*; D012486:Salmonella typhimurium / Q000187:drug effects* / Q000472:pathogenicity; D012772:Shock, Septic / Q000276:immunology / Q000382:microbiology / Q000473:pathology / Q000517:prevention & control*; D013154:Spleen / Q000187:drug effects / Q000473:pathology
D000900:Anti-Bacterial Agents; D016207:Cytokines; D007840:Latex; D010940:Plant Proteins