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Inhibitive effect of TAK-242 on Tenon's capsule fibroblasts proliferation in rat eyes
Liang Liang1,2, Meng-Nan Zhu1,2,3, Bao-Ji Chen1,2
1Department of Ophthalmology, The First College of Clinical Medical Science, China Three Gorges University, Yichang 443003, Hubei Province, China.
Aim:
To study the inhibition effect of TAK-242 on the proliferation of rat eye Tenon's capsule fibroblasts via the toll-like receptor 4 (TLR4) signaling pathway.
Methods:
SD rat Tenon's capsule fibroblasts were extracted and cultured, then the cells were divided into normal control group, lipopolysaccharide (LPS) group (10 g/mL LPS) and TAK-242 group (1 µmol/L TAK-242, and 10 µg/mL LPS after 30min). The expressions of TLR4, transforming growth factor-β1 (TGF-β1) and interleukin-6 (IL-6) in each group were detected by Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR). Cell proliferation was detected by cell counting kit-8 (CCK-8).
Results:
Double immunofluorescent labeling in the extracted cells showed negative keratin staining and positive vimentin staining. Western blot showed that the LPS group had the highest expression of TLR4 and TGF-β1 (P<0.01). Enzyme linked immunosorbent assay (ELISA) also showed that the secretion of IL-6 was the highest in LPS group (P<0.01). But there was no significant difference in TLR4 and TGF-1, as well as IL-6 expressions between the TAK-242 group and the normal control group (P>0.05). RT-PCR showed that the IL-6 mRNA expression in LPS group was the highest in the three groups (P<0.01).
Conclusion:
TAK-242 inhibits the proliferation of LPS-induced Tenon's capsule fibroblasts and the release of inflammatory factors by regulating the TLR4 signaling pathway, providing a new idea for reducing the scarring of the filter passage after glaucoma filtration surgery.
Insights
TAK-242 effectively inhibits the proliferation of rat Tenon
Area of Science:
- Immunology and Cell Biology
- Ophthalmology Research
Background:
- Tenon's capsule fibroblasts play a role in ocular wound healing and scarring.
- The toll-like receptor 4 (TLR4) signaling pathway is implicated in inflammatory responses.
- Glaucoma filtration surgery can lead to scarring, impacting surgical outcomes.
Purpose of the Study:
- To investigate the inhibitory effect of TAK-242 on rat eye Tenon's capsule fibroblast proliferation.
- To elucidate the role of the toll-like receptor 4 (TLR4) signaling pathway in this process.
Main Methods:
- Rat Tenon's capsule fibroblasts were cultured and treated with lipopolysaccharide (LPS) and TAK-242.
- Expression of TLR4, TGF-β1, and IL-6 was analyzed using Western blot and RT-PCR.
- Cell proliferation was assessed using the CCK-8 assay.
Main Results:
- LPS treatment significantly increased the expression of TLR4, TGF-β1, and IL-6.
- TAK-242 treatment, in combination with LPS, normalized the expression of these molecules compared to controls.
- TAK-242 inhibited LPS-induced fibroblast proliferation.
Conclusions:
- TAK-242 inhibits lipopolysaccharide-induced Tenon's capsule fibroblast proliferation via the TLR4 pathway.
- TAK-242 reduces the release of inflammatory factors, suggesting potential in mitigating post-surgical scarring.
- This study offers a potential therapeutic strategy for reducing scarring after glaucoma filtration surgery.
