Abstract:AIM: To investigate the effect of connective tissue growth factor(CTGF)on the proliferation and migration and transformation in Tenon's capsule fibroblasts(Tfb)of primary open angle glaucoma(POAG)patients.
METHODS: Tfb were cultured from Tenon's tissues in POAG patients in vitro. The free-serum DMEM-F12 containing 1.0, 10.0, 100.0ng/mL of CTGF was added into medium for 24h and 48h in different experimental group respectively, and only equal volume of free-serum DMEM-F12 was added in the negative control group. After 24h, the cell proliferation was analyzed through MTT assay, and migration was evaluated by crutch method. After 48h, Semi-quantitative RT-PCR was used to observed the mRNA expression of α-smooth muscl actin(α-SMA), and expression of α-SMA protein was examined by immunochemistry.
RESULTS: The proliferation values A of the cells in 1.0, 10.0, 100.0ng/mL of CTGF group respectively were 0.436±0.009, 0.643±0.009, 0.679±0.006, and 0.423 ±0.156 in the negative control group. The migrated cell number was 34.600±3.507, 70.400±2.074, 80.000±2.915 in different concentrations of CTGF group respectively, and 31.000±3.536 in the negative control group. And also in different experimental groups respectively, the absorbance ratio of α-SMA/β-actin was 0.873±0.161, 1.213±0.312, 1.352±0.376, and 0.851±0.158 in the negative control group, the expressing levels A of α-SMA protein in Tfb were 0.110±0.026, 0.141±0.017, 0.175±0.027, and 0.108±0.020 in the negative control group. The statistics of the above experimental data showed that, comparing with the negative control group, the 10.0 and 100.0ng/mL CTGF groups was statistically significant different(P<0.05), but there was no statistical different between the 1.0ng/mL CTGF group and the negative control group(P>0.05).
CONCLUSION: The proliferation, migration, and phenotypic transformation of Tfb can be promoted in CTGF group in POAG patients. These findings suggest that CTGF may play a role in the development of filtering bleb scarring.
