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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

Roscvovitine promotes apoptosis through increased expression of cleaved caspase 3 and decreased expression of Mcl-1. The expression of COX-2, phosphorylated NF-κB and Mcl-1 were increased at 24 h, but decreased gradually over time. The expression of COX-2 was significantly different between the LPS and roscovitine groups at 24 and 72 h (P=0.021 and 0.035, respectively). The expression of phosphorylated NF-κB was significantly decreased at 48 h (P=0.047). The expression of cleaved caspase 3 was markedly increased in the roscovitine group at 24 h (P=0.029). No statistically difference was observed in Mcl-1 expression among the groups. *P<0.05 vs. the LPS group. LPS, lipopolysaccharide; COX-2, cyclooxygenase-2; NF-κB, nuclear factor-κB; Mcl-1, myeloid cell leukemia 1.
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f5-mmr-14-02-1083: Roscvovitine promotes apoptosis through increased expression of cleaved caspase 3 and decreased expression of Mcl-1. The expression of COX-2, phosphorylated NF-κB and Mcl-1 were increased at 24 h, but decreased gradually over time. The expression of COX-2 was significantly different between the LPS and roscovitine groups at 24 and 72 h (P=0.021 and 0.035, respectively). The expression of phosphorylated NF-κB was significantly decreased at 48 h (P=0.047). The expression of cleaved caspase 3 was markedly increased in the roscovitine group at 24 h (P=0.029). No statistically difference was observed in Mcl-1 expression among the groups. *P<0.05 vs. the LPS group. LPS, lipopolysaccharide; COX-2, cyclooxygenase-2; NF-κB, nuclear factor-κB; Mcl-1, myeloid cell leukemia 1.

Mentions: Western blot analysis revealed that the protein expression level of COX-2 was markedly increased at 12 and 24 h; however, the level decreased to almost normal levels at 48 and 72 h in the four groups (Fig. 5). The expression of COX-2 was significantly lower in the roscovitine group compared with the control groups at 24 (P=0.021) and 72 h (P=0.035). A similar trend was observed in the levels of phosphorylated NF-κB p65, and compared with the LPS group, significantly less expression was observed in the roscovitine group at 48 h (P= 0.047).


Roscovitine ameliorates endotoxin-induced uveitis through neutrophil apoptosis.

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

Roscvovitine promotes apoptosis through increased expression of cleaved caspase 3 and decreased expression of Mcl-1. The expression of COX-2, phosphorylated NF-κB and Mcl-1 were increased at 24 h, but decreased gradually over time. The expression of COX-2 was significantly different between the LPS and roscovitine groups at 24 and 72 h (P=0.021 and 0.035, respectively). The expression of phosphorylated NF-κB was significantly decreased at 48 h (P=0.047). The expression of cleaved caspase 3 was markedly increased in the roscovitine group at 24 h (P=0.029). No statistically difference was observed in Mcl-1 expression among the groups. *P<0.05 vs. the LPS group. LPS, lipopolysaccharide; COX-2, cyclooxygenase-2; NF-κB, nuclear factor-κB; Mcl-1, myeloid cell leukemia 1.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5-mmr-14-02-1083: Roscvovitine promotes apoptosis through increased expression of cleaved caspase 3 and decreased expression of Mcl-1. The expression of COX-2, phosphorylated NF-κB and Mcl-1 were increased at 24 h, but decreased gradually over time. The expression of COX-2 was significantly different between the LPS and roscovitine groups at 24 and 72 h (P=0.021 and 0.035, respectively). The expression of phosphorylated NF-κB was significantly decreased at 48 h (P=0.047). The expression of cleaved caspase 3 was markedly increased in the roscovitine group at 24 h (P=0.029). No statistically difference was observed in Mcl-1 expression among the groups. *P<0.05 vs. the LPS group. LPS, lipopolysaccharide; COX-2, cyclooxygenase-2; NF-κB, nuclear factor-κB; Mcl-1, myeloid cell leukemia 1.
Mentions: Western blot analysis revealed that the protein expression level of COX-2 was markedly increased at 12 and 24 h; however, the level decreased to almost normal levels at 48 and 72 h in the four groups (Fig. 5). The expression of COX-2 was significantly lower in the roscovitine group compared with the control groups at 24 (P=0.021) and 72 h (P=0.035). A similar trend was observed in the levels of phosphorylated NF-κB p65, and compared with the LPS group, significantly less expression was observed in the roscovitine group at 48 h (P= 0.047).

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