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Intravitreal TSG-6 suppresses laser-induced choroidal neovascularization by inhibiting CCR2+ monocyte recruitment.

Kim SJ, Lee HJ, Yun JH, Ko JH, Choi da Y, Oh JY - Sci Rep (2015)

Bottom Line: An intravitreal injection of TSG-6 suppressed the expression of VEGF and pro-inflammatory cytokines including CCL2, and reduced the size of CNV.Also, the number of Iba(+) and CCR2(+) cells including infiltrating macrophages was markedly lower in the CNV lesion of TSG-6-treated eyes.Further analysis identified CCR2(+) CD11b(+) CD11c(+) cells and CCR2(+) CD11b(-)CD11c(+) cells as the cell populations that were increased by laser injury and reduced by TSG-6 treatment.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Ophthalmology, Samsung Medical Center, Sungkyunkwan University School of Medicine, 50 Irwon-dong, Gangnam-gu, Seoul, 135-710, Korea [2] Samsung Biomedical Research Institute, 50 Irwon-dong, Gangnam-gu, Seoul, 135-710, Korea.

ABSTRACT
Choroidal neovascularization (CNV) is the hallmark of wet age-related macular degeneration (AMD), one of the leading causes of blindness in the elderly. Although the pathogenesis of CNV is not clear, a number of studies show that ocular-infiltrating macrophages and inflammation play a critical role in the development of CNV. TNFα-stimulated gene/protein (TSG)-6 is a multifunctional endogenous protein that has anti-inflammatory activities partly by regulating macrophage activation. Therefore, we here investigated the therapeutic potential of TSG-6 in a rat model of CNV induced by laser photocoagulation. Time course analysis showed that the expression of VEGF and pro-inflammatory cytokines in the choroid was up-regulated early after laser injury, and gradually decreased to baseline over 14 days. An intravitreal injection of TSG-6 suppressed the expression of VEGF and pro-inflammatory cytokines including CCL2, and reduced the size of CNV. Also, the number of Iba(+) and CCR2(+) cells including infiltrating macrophages was markedly lower in the CNV lesion of TSG-6-treated eyes. Further analysis identified CCR2(+) CD11b(+) CD11c(+) cells and CCR2(+) CD11b(-)CD11c(+) cells as the cell populations that were increased by laser injury and reduced by TSG-6 treatment. Together, the results demonstrate that TSG-6 inhibits inflammation and CCR2(+) monocyte recruitment into the choroid, and suppresses the development of CNV.

No MeSH data available.


Related in: MedlinePlus

TSG-6 suppresses inflammatory responses in the choroid and retina after laser injury.(a) Real-time RT PCR of the RPE-choroidal tissue revealed that mRNA levels of TNF-α, IL-1β, and IL-6 were highly increased at day 1 after laser injury, and gradually decreased to baseline over 14 days. The levels of TNF-α, IL-1β, and IL-6 in the RPE-choroid were significantly lower in TSG-6-treated eyes at days 1, 3, and 7, compared to PBS-treated controls. (b) ELISA confirmed that the protein levels of IL-1β and IL-6 in the RPE-choroid were significantly reduced by TSG-6 treatment. (c) Real-time RT PCR of the retinal tissue showed that the mRNA levels of TNF-α, IL-1β, and IL-6 increased at day 1 after laser injury, and rapidly normalized until 7 days. The levels of TNF-α, IL-1β, and IL-6 transcripts in the retina were significantly lower in TSG-6-treated eyes at days 1 and 3. (d) Additional assay with ELISA confirmed that the levels of IL-1β and IL-6 proteins in the retina were significantly lower in TSG-6-treated group. Data are presented as mean±/+SD from three independent experiments. Each individual experiment included six rats per group at each time point. RQ means a ratio of mRNA levels relative to those in normal eyes. *p < 0.05, **p < 0.01.
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f2: TSG-6 suppresses inflammatory responses in the choroid and retina after laser injury.(a) Real-time RT PCR of the RPE-choroidal tissue revealed that mRNA levels of TNF-α, IL-1β, and IL-6 were highly increased at day 1 after laser injury, and gradually decreased to baseline over 14 days. The levels of TNF-α, IL-1β, and IL-6 in the RPE-choroid were significantly lower in TSG-6-treated eyes at days 1, 3, and 7, compared to PBS-treated controls. (b) ELISA confirmed that the protein levels of IL-1β and IL-6 in the RPE-choroid were significantly reduced by TSG-6 treatment. (c) Real-time RT PCR of the retinal tissue showed that the mRNA levels of TNF-α, IL-1β, and IL-6 increased at day 1 after laser injury, and rapidly normalized until 7 days. The levels of TNF-α, IL-1β, and IL-6 transcripts in the retina were significantly lower in TSG-6-treated eyes at days 1 and 3. (d) Additional assay with ELISA confirmed that the levels of IL-1β and IL-6 proteins in the retina were significantly lower in TSG-6-treated group. Data are presented as mean±/+SD from three independent experiments. Each individual experiment included six rats per group at each time point. RQ means a ratio of mRNA levels relative to those in normal eyes. *p < 0.05, **p < 0.01.

Mentions: We next evaluated the effects of TSG-6 on inflammatory responses in the RPE-choroid and retina after laser injury. Time course analysis revealed that the transcript levels of TNF-α, IL-1β, and IL-6 were highly increased in both RPE-choroid and retina at day 1 after injury, indicating that laser photocoagulation markedly induced inflammatory responses in the eye (Fig. 2a,c). The levels of pro-inflammatory cytokines were decreased to baseline over 14 days. An intravitreal injection of TSG-6 significantly reduced the transcript levels of TNF-α, IL-1β, and IL-6 in the RPE-choroid at days 1, 3, and 7, and in the retina at days 1 and 3 (Fig. 2a,c). Similar results were obtained with ELISA. The protein levels of IL-1β and IL-6 at day 3 were significantly lower in the RPE-choroid and retina of TSG-6-treated eyes compared to PBS-treated controls (Fig. 2b,d).


Intravitreal TSG-6 suppresses laser-induced choroidal neovascularization by inhibiting CCR2+ monocyte recruitment.

Kim SJ, Lee HJ, Yun JH, Ko JH, Choi da Y, Oh JY - Sci Rep (2015)

TSG-6 suppresses inflammatory responses in the choroid and retina after laser injury.(a) Real-time RT PCR of the RPE-choroidal tissue revealed that mRNA levels of TNF-α, IL-1β, and IL-6 were highly increased at day 1 after laser injury, and gradually decreased to baseline over 14 days. The levels of TNF-α, IL-1β, and IL-6 in the RPE-choroid were significantly lower in TSG-6-treated eyes at days 1, 3, and 7, compared to PBS-treated controls. (b) ELISA confirmed that the protein levels of IL-1β and IL-6 in the RPE-choroid were significantly reduced by TSG-6 treatment. (c) Real-time RT PCR of the retinal tissue showed that the mRNA levels of TNF-α, IL-1β, and IL-6 increased at day 1 after laser injury, and rapidly normalized until 7 days. The levels of TNF-α, IL-1β, and IL-6 transcripts in the retina were significantly lower in TSG-6-treated eyes at days 1 and 3. (d) Additional assay with ELISA confirmed that the levels of IL-1β and IL-6 proteins in the retina were significantly lower in TSG-6-treated group. Data are presented as mean±/+SD from three independent experiments. Each individual experiment included six rats per group at each time point. RQ means a ratio of mRNA levels relative to those in normal eyes. *p < 0.05, **p < 0.01.
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f2: TSG-6 suppresses inflammatory responses in the choroid and retina after laser injury.(a) Real-time RT PCR of the RPE-choroidal tissue revealed that mRNA levels of TNF-α, IL-1β, and IL-6 were highly increased at day 1 after laser injury, and gradually decreased to baseline over 14 days. The levels of TNF-α, IL-1β, and IL-6 in the RPE-choroid were significantly lower in TSG-6-treated eyes at days 1, 3, and 7, compared to PBS-treated controls. (b) ELISA confirmed that the protein levels of IL-1β and IL-6 in the RPE-choroid were significantly reduced by TSG-6 treatment. (c) Real-time RT PCR of the retinal tissue showed that the mRNA levels of TNF-α, IL-1β, and IL-6 increased at day 1 after laser injury, and rapidly normalized until 7 days. The levels of TNF-α, IL-1β, and IL-6 transcripts in the retina were significantly lower in TSG-6-treated eyes at days 1 and 3. (d) Additional assay with ELISA confirmed that the levels of IL-1β and IL-6 proteins in the retina were significantly lower in TSG-6-treated group. Data are presented as mean±/+SD from three independent experiments. Each individual experiment included six rats per group at each time point. RQ means a ratio of mRNA levels relative to those in normal eyes. *p < 0.05, **p < 0.01.
Mentions: We next evaluated the effects of TSG-6 on inflammatory responses in the RPE-choroid and retina after laser injury. Time course analysis revealed that the transcript levels of TNF-α, IL-1β, and IL-6 were highly increased in both RPE-choroid and retina at day 1 after injury, indicating that laser photocoagulation markedly induced inflammatory responses in the eye (Fig. 2a,c). The levels of pro-inflammatory cytokines were decreased to baseline over 14 days. An intravitreal injection of TSG-6 significantly reduced the transcript levels of TNF-α, IL-1β, and IL-6 in the RPE-choroid at days 1, 3, and 7, and in the retina at days 1 and 3 (Fig. 2a,c). Similar results were obtained with ELISA. The protein levels of IL-1β and IL-6 at day 3 were significantly lower in the RPE-choroid and retina of TSG-6-treated eyes compared to PBS-treated controls (Fig. 2b,d).

Bottom Line: An intravitreal injection of TSG-6 suppressed the expression of VEGF and pro-inflammatory cytokines including CCL2, and reduced the size of CNV.Also, the number of Iba(+) and CCR2(+) cells including infiltrating macrophages was markedly lower in the CNV lesion of TSG-6-treated eyes.Further analysis identified CCR2(+) CD11b(+) CD11c(+) cells and CCR2(+) CD11b(-)CD11c(+) cells as the cell populations that were increased by laser injury and reduced by TSG-6 treatment.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Ophthalmology, Samsung Medical Center, Sungkyunkwan University School of Medicine, 50 Irwon-dong, Gangnam-gu, Seoul, 135-710, Korea [2] Samsung Biomedical Research Institute, 50 Irwon-dong, Gangnam-gu, Seoul, 135-710, Korea.

ABSTRACT
Choroidal neovascularization (CNV) is the hallmark of wet age-related macular degeneration (AMD), one of the leading causes of blindness in the elderly. Although the pathogenesis of CNV is not clear, a number of studies show that ocular-infiltrating macrophages and inflammation play a critical role in the development of CNV. TNFα-stimulated gene/protein (TSG)-6 is a multifunctional endogenous protein that has anti-inflammatory activities partly by regulating macrophage activation. Therefore, we here investigated the therapeutic potential of TSG-6 in a rat model of CNV induced by laser photocoagulation. Time course analysis showed that the expression of VEGF and pro-inflammatory cytokines in the choroid was up-regulated early after laser injury, and gradually decreased to baseline over 14 days. An intravitreal injection of TSG-6 suppressed the expression of VEGF and pro-inflammatory cytokines including CCL2, and reduced the size of CNV. Also, the number of Iba(+) and CCR2(+) cells including infiltrating macrophages was markedly lower in the CNV lesion of TSG-6-treated eyes. Further analysis identified CCR2(+) CD11b(+) CD11c(+) cells and CCR2(+) CD11b(-)CD11c(+) cells as the cell populations that were increased by laser injury and reduced by TSG-6 treatment. Together, the results demonstrate that TSG-6 inhibits inflammation and CCR2(+) monocyte recruitment into the choroid, and suppresses the development of CNV.

No MeSH data available.


Related in: MedlinePlus