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Roscovitine ameliorates endotoxin-induced uveitis through neutrophil apoptosis.

Jiang ZX, Qiu S, Lou BS, Yang Y, Wang WC, Lin XF - Mol Med Rep (2016)

Bottom Line: The number of infiltrating cells was similar in the LPS and LPS plus vehicle groups, while there were significantly less in the roscovitine group at 24 h.This trend in mRNA expression was similar in the LPS and LPS plus vehicle groups; however, the expression levels decreased more quickly in the roscovitine group at 24 and 48 h.In conclusion, roscovitine ameliorates EIU by effecting neutrophil apoptosis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat‑Sen University, Guangzhou, Guangdong 510060, P.R. China.

ABSTRACT
Neutrophils have been recognized as critical response cells during the pathogenesis of endotoxin‑induced uveitis (EIU). Apoptosis of neutrophils induced by roscovitine has previously been demonstrated to ameliorate inflammation in several in vivo models. The present study aimed to assess whether roscovitine ameliorates EIU. EIU was induced in female C57BL/6 mice by a single intravitreal injection of lipopolysaccharide (LPS; 250 ng). The mice were divided into three groups as follows: LPS alone, LPS plus vehicle, LPS plus roscovitine (50 mg/kg). The mice were euthanized 12, 24, 48 and 72 h after LPS‑induced uveitis. Accumulation of inflammatory cells in the vitreous body was confirmed by immunohistochemistry, and quantified following hematoxylin and eosin staining. Terminal deoxynucleotidyl transferase dUTP nick‑end labeling was performed to detect of apoptotic cells. The mRNA levels of inflammatory cytokines were analyzed by reverse transcription‑quantitative polymerase chain reaction and the changes in protein levels were analyzed by western blotting. Inflammatory cells accumulated in the vitreous near the optic nerve head and the quantity peaked at 24 h after LPS injection. Immunohistochemistry revealed that the majority of the inflammatory cells were neutrophils. The number of infiltrating cells was similar in the LPS and LPS plus vehicle groups, while there were significantly less in the roscovitine group at 24 h. Apoptosis of neutrophils was observed between 12 and 48 h after roscovitine injection, while no apoptosis was observed in the other groups. The mRNA expression levels of GMCSF, CINC‑1 and ICAM‑1 peaked at 12 h after LPS injection, and decreased to normal levels at 72 h. This trend in mRNA expression was similar in the LPS and LPS plus vehicle groups; however, the expression levels decreased more quickly in the roscovitine group at 24 and 48 h. Following roscovitine administration, upregulated cleaved caspase 3 expression levels and downregulated Mcl‑1 expression levels were observed. In conclusion, roscovitine ameliorates EIU by effecting neutrophil apoptosis. Timely apoptosis of neutrophils may be an effective process to promote the amelioration of EIU.

No MeSH data available.


Related in: MedlinePlus

Reverse transcription-quantitative polymerase chain reaction analysis of mRNA expression for inflammatory cytokines. The relative expression level was similar in the LPS and LPS plus vehicle groups at the identical time points, whereas the relative mRNA expression levels were lower in the roscovitine group at the respective time point. The results for IL-1 were significant at 24 and 48 h when compared with the LPS group (P= 0.042 and 0.033, respectively). The difference of GMCSF was significant at 24 h when compared with LPS groups (P=0.026). *P<0.05 vs. the LPS group. LPS, lipopolysaccharide; IL-1, interleukin-1; GMCSF, granulocyte-macrophage colony-stimulating factor; CINC-1, cytokine-induced neutrophil chemoattractant 1; ICAM-1, intercellular adhesion molecule 1.
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f4-mmr-14-02-1083: Reverse transcription-quantitative polymerase chain reaction analysis of mRNA expression for inflammatory cytokines. The relative expression level was similar in the LPS and LPS plus vehicle groups at the identical time points, whereas the relative mRNA expression levels were lower in the roscovitine group at the respective time point. The results for IL-1 were significant at 24 and 48 h when compared with the LPS group (P= 0.042 and 0.033, respectively). The difference of GMCSF was significant at 24 h when compared with LPS groups (P=0.026). *P<0.05 vs. the LPS group. LPS, lipopolysaccharide; IL-1, interleukin-1; GMCSF, granulocyte-macrophage colony-stimulating factor; CINC-1, cytokine-induced neutrophil chemoattractant 1; ICAM-1, intercellular adhesion molecule 1.

Mentions: RT-qPCR analysis demonstrated that the mRNA expression level of interleukin 1 increased, peaked at 24 h after LPS injection, and decreased gradually by 72 h. The mRNA expression levels of granulocyte-macrophage colony-stimulating factor (GMCSF), cytokine-induced neutrophil chemoattractant 1 (CINC-1) and intercellular adhesion molecule 1 (ICAM-1) peaked at 12 h, then decreased rapidly to the normal levels (Fig. 4). The relative mRNA levels were similar in the LPS and LPS plus vehicle groups at the identical time points, while the relative levels were lower in the roscovitine group at the respective time points. In particular, the mRNA levels of interleukin 1 in the roscovitine groups were significantly reduced compared with the LPS group at 24 (P=0.042) and 48 h (P=0.033).


Roscovitine ameliorates endotoxin-induced uveitis through neutrophil apoptosis.

Jiang ZX, Qiu S, Lou BS, Yang Y, Wang WC, Lin XF - Mol Med Rep (2016)

Reverse transcription-quantitative polymerase chain reaction analysis of mRNA expression for inflammatory cytokines. The relative expression level was similar in the LPS and LPS plus vehicle groups at the identical time points, whereas the relative mRNA expression levels were lower in the roscovitine group at the respective time point. The results for IL-1 were significant at 24 and 48 h when compared with the LPS group (P= 0.042 and 0.033, respectively). The difference of GMCSF was significant at 24 h when compared with LPS groups (P=0.026). *P<0.05 vs. the LPS group. LPS, lipopolysaccharide; IL-1, interleukin-1; GMCSF, granulocyte-macrophage colony-stimulating factor; CINC-1, cytokine-induced neutrophil chemoattractant 1; ICAM-1, intercellular adhesion molecule 1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940085&req=5

f4-mmr-14-02-1083: Reverse transcription-quantitative polymerase chain reaction analysis of mRNA expression for inflammatory cytokines. The relative expression level was similar in the LPS and LPS plus vehicle groups at the identical time points, whereas the relative mRNA expression levels were lower in the roscovitine group at the respective time point. The results for IL-1 were significant at 24 and 48 h when compared with the LPS group (P= 0.042 and 0.033, respectively). The difference of GMCSF was significant at 24 h when compared with LPS groups (P=0.026). *P<0.05 vs. the LPS group. LPS, lipopolysaccharide; IL-1, interleukin-1; GMCSF, granulocyte-macrophage colony-stimulating factor; CINC-1, cytokine-induced neutrophil chemoattractant 1; ICAM-1, intercellular adhesion molecule 1.
Mentions: RT-qPCR analysis demonstrated that the mRNA expression level of interleukin 1 increased, peaked at 24 h after LPS injection, and decreased gradually by 72 h. The mRNA expression levels of granulocyte-macrophage colony-stimulating factor (GMCSF), cytokine-induced neutrophil chemoattractant 1 (CINC-1) and intercellular adhesion molecule 1 (ICAM-1) peaked at 12 h, then decreased rapidly to the normal levels (Fig. 4). The relative mRNA levels were similar in the LPS and LPS plus vehicle groups at the identical time points, while the relative levels were lower in the roscovitine group at the respective time points. In particular, the mRNA levels of interleukin 1 in the roscovitine groups were significantly reduced compared with the LPS group at 24 (P=0.042) and 48 h (P=0.033).

Bottom Line: The number of infiltrating cells was similar in the LPS and LPS plus vehicle groups, while there were significantly less in the roscovitine group at 24 h.This trend in mRNA expression was similar in the LPS and LPS plus vehicle groups; however, the expression levels decreased more quickly in the roscovitine group at 24 and 48 h.In conclusion, roscovitine ameliorates EIU by effecting neutrophil apoptosis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat‑Sen University, Guangzhou, Guangdong 510060, P.R. China.

ABSTRACT
Neutrophils have been recognized as critical response cells during the pathogenesis of endotoxin‑induced uveitis (EIU). Apoptosis of neutrophils induced by roscovitine has previously been demonstrated to ameliorate inflammation in several in vivo models. The present study aimed to assess whether roscovitine ameliorates EIU. EIU was induced in female C57BL/6 mice by a single intravitreal injection of lipopolysaccharide (LPS; 250 ng). The mice were divided into three groups as follows: LPS alone, LPS plus vehicle, LPS plus roscovitine (50 mg/kg). The mice were euthanized 12, 24, 48 and 72 h after LPS‑induced uveitis. Accumulation of inflammatory cells in the vitreous body was confirmed by immunohistochemistry, and quantified following hematoxylin and eosin staining. Terminal deoxynucleotidyl transferase dUTP nick‑end labeling was performed to detect of apoptotic cells. The mRNA levels of inflammatory cytokines were analyzed by reverse transcription‑quantitative polymerase chain reaction and the changes in protein levels were analyzed by western blotting. Inflammatory cells accumulated in the vitreous near the optic nerve head and the quantity peaked at 24 h after LPS injection. Immunohistochemistry revealed that the majority of the inflammatory cells were neutrophils. The number of infiltrating cells was similar in the LPS and LPS plus vehicle groups, while there were significantly less in the roscovitine group at 24 h. Apoptosis of neutrophils was observed between 12 and 48 h after roscovitine injection, while no apoptosis was observed in the other groups. The mRNA expression levels of GMCSF, CINC‑1 and ICAM‑1 peaked at 12 h after LPS injection, and decreased to normal levels at 72 h. This trend in mRNA expression was similar in the LPS and LPS plus vehicle groups; however, the expression levels decreased more quickly in the roscovitine group at 24 and 48 h. Following roscovitine administration, upregulated cleaved caspase 3 expression levels and downregulated Mcl‑1 expression levels were observed. In conclusion, roscovitine ameliorates EIU by effecting neutrophil apoptosis. Timely apoptosis of neutrophils may be an effective process to promote the amelioration of EIU.

No MeSH data available.


Related in: MedlinePlus