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[The effect of nerve growth factor on proliferation and expression of mucin gene in human conjunctival goblet cells]
1Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Capital University of Medical Sciences, Beijing, China.
Objective:
To study the effect of nerve growth factor (NGF) on the proliferation and the expression of MUC5AC gene in cultured human conjunctival goblet cells.
Methods:
The presence of high affinity receptors of nerve growth factor (TrkA) in cultured human conjunctival goblet cells was identified with immunocytochemical analysis. Proliferation of goblet cells was determined by MTT proliferation assay after exposure to different concentrations of NGF, (20, 40, 60 and 80 ng/ml) in serum-free RPMI-1640 culture medium (RPMI) for 1, 3 and 5 days. The effect of NGF on gene expression of MUC5AC of goblet cells was determined by RT-PCR after exposing cells to NGF (80 ng/ml in serum-free RPMI) for 5 days.
Results:
Human conjunctival goblet cells in subculture exhibited positive staining for TrkA. After exposure to NGF, the differences in absorption value of MTT test of goblet cells cultured with different levels of NGF (F = 12.766, P = 0.000) and at different time points (F = 892.406, P = 0.000) were statistically significant, but the difference in interaction effects was not significant. MUC5AC gene expression of NGF group was higher than that of the control group (t = 4.963, P = 0.000).
Conclusions:
NGF receptors (TrkA) are present in human conjunctival goblet cells. NGF causes a dose-dependent increase in the proliferation of goblet cells. The expression of MUC5AC gene in goblet cells is upgraded by NGF.
Insights
Nerve growth factor (NGF) promotes the proliferation of human conjunctival goblet cells and increases MUC5AC gene expression, indicating its potential role in ocular surface health.
Area of Science:
- Ophthalmology
- Cell Biology
- Molecular Biology
Context:
- Human conjunctival goblet cells are crucial for ocular surface lubrication and protection.
- Nerve growth factor (NGF) is known to play roles in neuronal development and survival.
- The specific effects of NGF on conjunctival goblet cells, particularly MUC5AC gene expression, were not well understood.
Purpose:
- To investigate the presence of NGF receptors (TrkA) in human conjunctival goblet cells.
- To determine the effect of NGF on the proliferation of these cells.
- To analyze the impact of NGF on MUC5AC gene expression in conjunctival goblet cells.
Summary:
- Immunocytochemical analysis confirmed the presence of TrkA receptors on human conjunctival goblet cells.
- MTT assays revealed that NGF significantly increases goblet cell proliferation in a dose-dependent manner.
- RT-PCR demonstrated that NGF upregulates the expression of the MUC5AC gene in these cells.
Impact:
- This study identifies NGF as a potential therapeutic agent for conditions involving goblet cell dysfunction.
- The findings suggest NGF's role in maintaining ocular surface homeostasis through enhanced mucus production and cell proliferation.
- Further research into NGF-based therapies could offer new strategies for treating dry eye disease and other ocular surface disorders.
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