Knockdown of TNF-alpha alleviates acute lung injury in rats with intestinal ischemia and reperfusion injury by upregulating IL-10 expression

摘要

Intestinal ischemia and reperfusion (II/R) injury often triggers severe injury in remote organs, with the lungs being considered the main target. Excessive elevation of proinflammatory cytokines is a major contributor in the occurrence and development of II/R-induced acute lung injury (ALI). Therefore, the present study aimed to investigate whether blocking tumor necrosis factor-alpha (TNF-alpha) expression could protect the lungs from injury following II/R, and to explore the possible underlying mechanism involving interleukin-10 (IL-10). Briefly, II/R was induced in rats by 40 min occlusion of the superior mesenteric artery and celiac artery, followed by 8, 16 or 24 h of reperfusion. Subsequently, lentiviral vectors containing TNF-alpha short hairpin (sh)RNA were injected into the right lung tissues, in order to induce TNF-alpha knockdown. The severity of ALI was determined according to lung injury scores and lung edema (lung wet/dry weight ratio). The expression levels of TNF-alpha were analyzed by quantitative polymerase chain reaction (qPCR), western blotting and immunofluorescence (IF) staining. IL-10 expression, in response to TNF-alpha knockdown, was detected in lung tissues by qPCR and IF. The results detected marked inflammatory responses, and increased levels of lung wet/dry weight ratio and TNF-alpha expression, in the lungs of II/R rats. Conversely, treatment with TNF-alpha shRNA significantly alleviated the severity of ALI and upregulated the expression levels of IL-10 in lung tissues. These findings suggested that TNF-alpha RNA interference may exert a protective effect on II/R-induced ALI via the upregulation of IL-10. Therefore, TNF-alpha knockdown may be considered a potential strategy for the prevention or treatment of ALI induced by II/R in future clinical trials.