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JNK inhibitor SP600125 protects against lipopolysaccharide-induced acute lung injury via upregulation of claudin-4.

Zheng Y, Zhang M, Zhao Y, Chen J, Li B, Cai W - Exp Ther Med (2014)

Bottom Line: Pulmonary edema, the expression of inflammatory cytokines and pathological alterations were found to be significantly attenuated in LPS-induced ALI following treatment with SP600125 in vivo.Furthermore, flow cytometric analysis demonstrated that the apoptotic rate was significantly reduced in a concentration-dependent manner following SP600125 injection.In combination, the results from the present study indicated that the JNK inhibitor SP600125 protected against LPS-induced ALI in vivo and in vitro, possibly by upregulating the expression of claudin-4.

View Article: PubMed Central - PubMed

Affiliation: Department of Emergency, Zhejiang Provincial People's Hospital, Hangzhou, Zhejiang 310014, P.R. China.

ABSTRACT
Although in vitro studies have previously demonstrated that mitogen-activated protein kinases are important for the activation of transcription factors and the regulation of proinflammatory mediators, the function of c-Jun NH2-terminal kinase (JNK) in acute lung injury (ALI) remains to be fully elucidated. The present study aimed to investigate the effect of the JNK selective inhibitor SP600125 on lipopolysaccharide (LPS)-induced ALI. Pulmonary edema, the expression of inflammatory cytokines and pathological alterations were found to be significantly attenuated in LPS-induced ALI following treatment with SP600125 in vivo. In vitro, it was demonstrated that SP600125 administration significantly improved A549 cell viability in a dose-dependent manner using the Cell Counting kit-8 and the 5-ethynyl-2'-deoxyuridine incorporation assay. Furthermore, flow cytometric analysis demonstrated that the apoptotic rate was significantly reduced in a concentration-dependent manner following SP600125 injection. At the molecular level, SP600125 treatment dose-dependently inhibited JNK activation and upregulated claudin-4 expression in vivo and in vitro. In combination, the results from the present study indicated that the JNK inhibitor SP600125 protected against LPS-induced ALI in vivo and in vitro, possibly by upregulating the expression of claudin-4.

No MeSH data available.


Related in: MedlinePlus

SP600125 attenuates LPS-induced ALI in rats in vivo. (A) Following LPS injection with or without SP600125 treatment, the rats were sacrificed and their left lower lungs were obtained in order to determine the wet/dry weight ratio. *P<0.05 vs. control; #P<0.05 vs. ALI. (B and C) An enzyme-linked immunosorbent assay was performed to determine the expression of TNF-α and IL-6 in the bronchoalveolar lavage fluid in the rats from each group. *P<0.05 vs. control; #P<0.05 vs. ALI. (D) Lung tissue sections were stained using hematoxylin and eosin to determine the pathological alterations with or without SP600125 treatment. Representative images are shown from each group (magnification, ×200). Column 1 and 2 show different views in the same group. ALI, acute lung injury; NS, normal saline; JNK, c-Jun NH2-terminal kinase; IL-6, interleukin-6; TNF-α, tumor necrosis factor-α; LPS, lipopolysaccharide.
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f1-etm-08-01-0153: SP600125 attenuates LPS-induced ALI in rats in vivo. (A) Following LPS injection with or without SP600125 treatment, the rats were sacrificed and their left lower lungs were obtained in order to determine the wet/dry weight ratio. *P<0.05 vs. control; #P<0.05 vs. ALI. (B and C) An enzyme-linked immunosorbent assay was performed to determine the expression of TNF-α and IL-6 in the bronchoalveolar lavage fluid in the rats from each group. *P<0.05 vs. control; #P<0.05 vs. ALI. (D) Lung tissue sections were stained using hematoxylin and eosin to determine the pathological alterations with or without SP600125 treatment. Representative images are shown from each group (magnification, ×200). Column 1 and 2 show different views in the same group. ALI, acute lung injury; NS, normal saline; JNK, c-Jun NH2-terminal kinase; IL-6, interleukin-6; TNF-α, tumor necrosis factor-α; LPS, lipopolysaccharide.

Mentions: The lung W/D ratio was analyzed to evaluate pulmonary edema. The LPS-treated rats had higher W/D ratios compared with the control rats. However, the W/D ratio was significantly decreased following administration of SP600125 (Fig. 1A). Furthermore, the results from the ELISA demonstrated that the expression of TNF-α and IL-6 in the bronchoalveolar lavage fluid (BALF) in the LPS-treated rats was markedly increased compared with the rats in the control group. However, the expression levels of TNF-α and IL-6 in the BALF in rats in the SP600125 group were significantly decreased (Fig. 1B and C). To assess the pathological alterations, H&E staining was performed and the results revealed evidence of infiltration of inflammatory cells, interstitial edema and interalveolar septal thickening, as well as intra-alveolar and interstitial hemorrhage. However, following treatment with SP600125, the pathological changes in the lung tissues of the rats markedly decreased (Fig. 1D). These results demonstrated that SP600125 treatment alleviated LPS-induced ALI in vivo.


JNK inhibitor SP600125 protects against lipopolysaccharide-induced acute lung injury via upregulation of claudin-4.

Zheng Y, Zhang M, Zhao Y, Chen J, Li B, Cai W - Exp Ther Med (2014)

SP600125 attenuates LPS-induced ALI in rats in vivo. (A) Following LPS injection with or without SP600125 treatment, the rats were sacrificed and their left lower lungs were obtained in order to determine the wet/dry weight ratio. *P<0.05 vs. control; #P<0.05 vs. ALI. (B and C) An enzyme-linked immunosorbent assay was performed to determine the expression of TNF-α and IL-6 in the bronchoalveolar lavage fluid in the rats from each group. *P<0.05 vs. control; #P<0.05 vs. ALI. (D) Lung tissue sections were stained using hematoxylin and eosin to determine the pathological alterations with or without SP600125 treatment. Representative images are shown from each group (magnification, ×200). Column 1 and 2 show different views in the same group. ALI, acute lung injury; NS, normal saline; JNK, c-Jun NH2-terminal kinase; IL-6, interleukin-6; TNF-α, tumor necrosis factor-α; LPS, lipopolysaccharide.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4061205&req=5

f1-etm-08-01-0153: SP600125 attenuates LPS-induced ALI in rats in vivo. (A) Following LPS injection with or without SP600125 treatment, the rats were sacrificed and their left lower lungs were obtained in order to determine the wet/dry weight ratio. *P<0.05 vs. control; #P<0.05 vs. ALI. (B and C) An enzyme-linked immunosorbent assay was performed to determine the expression of TNF-α and IL-6 in the bronchoalveolar lavage fluid in the rats from each group. *P<0.05 vs. control; #P<0.05 vs. ALI. (D) Lung tissue sections were stained using hematoxylin and eosin to determine the pathological alterations with or without SP600125 treatment. Representative images are shown from each group (magnification, ×200). Column 1 and 2 show different views in the same group. ALI, acute lung injury; NS, normal saline; JNK, c-Jun NH2-terminal kinase; IL-6, interleukin-6; TNF-α, tumor necrosis factor-α; LPS, lipopolysaccharide.
Mentions: The lung W/D ratio was analyzed to evaluate pulmonary edema. The LPS-treated rats had higher W/D ratios compared with the control rats. However, the W/D ratio was significantly decreased following administration of SP600125 (Fig. 1A). Furthermore, the results from the ELISA demonstrated that the expression of TNF-α and IL-6 in the bronchoalveolar lavage fluid (BALF) in the LPS-treated rats was markedly increased compared with the rats in the control group. However, the expression levels of TNF-α and IL-6 in the BALF in rats in the SP600125 group were significantly decreased (Fig. 1B and C). To assess the pathological alterations, H&E staining was performed and the results revealed evidence of infiltration of inflammatory cells, interstitial edema and interalveolar septal thickening, as well as intra-alveolar and interstitial hemorrhage. However, following treatment with SP600125, the pathological changes in the lung tissues of the rats markedly decreased (Fig. 1D). These results demonstrated that SP600125 treatment alleviated LPS-induced ALI in vivo.

Bottom Line: Pulmonary edema, the expression of inflammatory cytokines and pathological alterations were found to be significantly attenuated in LPS-induced ALI following treatment with SP600125 in vivo.Furthermore, flow cytometric analysis demonstrated that the apoptotic rate was significantly reduced in a concentration-dependent manner following SP600125 injection.In combination, the results from the present study indicated that the JNK inhibitor SP600125 protected against LPS-induced ALI in vivo and in vitro, possibly by upregulating the expression of claudin-4.

View Article: PubMed Central - PubMed

Affiliation: Department of Emergency, Zhejiang Provincial People's Hospital, Hangzhou, Zhejiang 310014, P.R. China.

ABSTRACT
Although in vitro studies have previously demonstrated that mitogen-activated protein kinases are important for the activation of transcription factors and the regulation of proinflammatory mediators, the function of c-Jun NH2-terminal kinase (JNK) in acute lung injury (ALI) remains to be fully elucidated. The present study aimed to investigate the effect of the JNK selective inhibitor SP600125 on lipopolysaccharide (LPS)-induced ALI. Pulmonary edema, the expression of inflammatory cytokines and pathological alterations were found to be significantly attenuated in LPS-induced ALI following treatment with SP600125 in vivo. In vitro, it was demonstrated that SP600125 administration significantly improved A549 cell viability in a dose-dependent manner using the Cell Counting kit-8 and the 5-ethynyl-2'-deoxyuridine incorporation assay. Furthermore, flow cytometric analysis demonstrated that the apoptotic rate was significantly reduced in a concentration-dependent manner following SP600125 injection. At the molecular level, SP600125 treatment dose-dependently inhibited JNK activation and upregulated claudin-4 expression in vivo and in vitro. In combination, the results from the present study indicated that the JNK inhibitor SP600125 protected against LPS-induced ALI in vivo and in vitro, possibly by upregulating the expression of claudin-4.

No MeSH data available.


Related in: MedlinePlus