Abstract | OBJECTIVE: DESIGN: Short-chain antisense oligonucleotide molecules were designed with the aid of computer software programs, and their in vitro efficacies were assessed in cell culture systems based on inhibition of target protein expression. The in vivo efficacies were determined in intact sepsis rats using 35-day survival rate as a primary efficacy end point. SETTING: Animal research laboratory at a university. SUBJECTS: Male Sprague-Dawley rats (180-200 g). INTERVENTIONS: MEASUREMENTS AND MAIN RESULTS: In cell culture systems, 21 of the 105 antisense constructs were found to be efficacious in inhibiting secretory phospholipase A(2) IIa and cytosolic phospholipase A(2) IVa protein expression. In sepsis rats, antisense oligonucleotides were capable of reducing their target protein expression by 18%-61% in major organs such as liver, heart, and kidney. In animal experiments, sepsis without any treatment (Group 1) had a median survival time of 2 days and a zero (0) percent survival rate at day 14. Sepsis with antibiotic treatment (Group 2) had a median survival time of 6 days and a 35-day survival rate of 28%. Sepsis with cotreatment of antibiotics and antisense oligonucleotides (one against secretory phospholipase A2 IIa and the other against cytosolic phospholipase A(2) IVa) (Group 4) increased the median survival time from 6 to 35 days and the 35-day survival rate from 28% to 58.8% as compared with antibiotics alone (Group 4 vs. Group 2; p <.05). Sepsis with cotreatment of antibiotics and mismatch oligonucleotides (Group 3) did not affect the median survival time and the 35-day survival rate as compared to antibiotics alone (Group 3 vs. Group 2; p >.05). CONCLUSIONS: The results demonstrate that antisense strategy against secretory phospholipase A(2) IIa and cytosolic phospholipase A(2) IVa can inhibit their target protein expression in major organs and greatly improve the clinical outcome, i.e., an absolute reduction in 35-day mortality of 30.8%, in rats with sepsis. Our studies, thus, provide an improved method for the treatment of sepsis by targeting multiple forms of phospholipase A(2) isoenzymes with DNA antisense oligomers.
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Authors | Maw-Shung Liu, Chia-Hsiung Liu, Guang Wu, Yuefang Zhou |
Journal | Critical care medicine
(Crit Care Med)
Vol. 40
Issue 7
Pg. 2132-40
(Jul 2012)
ISSN: 1530-0293 [Electronic] United States |
PMID | 22564957
(Publication Type: Journal Article)
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Chemical References |
- Anti-Bacterial Agents
- Oligonucleotides, Antisense
- Group IV Phospholipases A2
- Phospholipases A2, Secretory
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Topics |
- Animals
- Anti-Bacterial Agents
(pharmacology)
- Blotting, Western
- Cell Line
- Disease Models, Animal
- Down-Regulation
- Group IV Phospholipases A2
(antagonists & inhibitors, metabolism)
- Heart
(drug effects)
- Kidney
(drug effects, metabolism)
- Liver
(drug effects, metabolism)
- Male
- Myocardium
(metabolism)
- Oligonucleotides, Antisense
(pharmacology)
- Phospholipases A2, Secretory
(antagonists & inhibitors, metabolism)
- Rats
- Rats, Sprague-Dawley
- Sepsis
(drug therapy, mortality)
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