advertisement
Abstract #19761 Published in IGR 9-4
Effects of NF-κB on proliferation and DNA synthesis in human trabecular meshwork cells
Yuan H; He XChinese Ophthalmic Research 2007; 25: 596-598
OBJECTIVE: Nuclear factor-κB (NF-κB) regulates the transcription of a wide array of gene products that are involved in many pathological and physiological pathways. Activation of NF-κB may be a pivotal event in proinflammatory signal transduction. Inflammatory responses commonly occur during the glaucoma onset, development and treatment, especially in that caused by trauma and uveitis. Our study was to explore the effects of NF-κB on proliferation and DNA synthesis in human trabecular meshwork cells. METHODS: Human trabecular meshwork cells were obtained from the donor eyes and cultivated in DMEM medium containing 15% FBS. The cells were divided into three groups: the normal control group, the control plasmid group (P65-C), and the pRSV-RelA/P65 plasmid group (P65). The lipofectin method was used to transfer P65 plasmid into the primary culture human trabecular meshwork cells in the P65 group. The proliferation and DNA synthesis of human trabecular meshwork cells were detected by MTT and 3H-TdR 24 hours after transfer. RESULTS: Human trabecular meshwork cells were successfully cultured in DMEM medium. Immunochemical analysis revealed human trabecular meshwork cells reacted positive to LN, FN, and collagen IV, but negative to VIII factor relevant antigen. At 24 hours after P65 gene transfer, the expression of P65 was dramatically elevated by Western blot (0.66 ± 0.12, 0.70 ± 0.08, 1.03 ± 0.02 in the control group, the P65-C group and the P65 group, respectively). MTT and 3H-TdR showed that the proliferation and DNA synthesis in human trabecular meshwork cells were suppressed after P65 plasmid transfer, displaying a significant difference compared with the control groups (P < 0. 05). CONCLUSION: Overexpression of P65 suppresses proliferation and DNA synthesis in human trabecular meshwork cells, suggesting the excess activation of NF-κB may be the important event to result in secondary glaucoma. LA: Chinese
Dr. H. Yuan, Department of Ophthalmology, Daping Hospital, Third Military Medical University, Chongqing 630000, China. yhf871@yahoo.com.cn
Classification:
3.5 Molecular biology incl. SiRNA (Part of: 3 Laboratory methods)
3.6 Cellular biology (Part of: 3 Laboratory methods)
2.5.1 Trabecular meshwork (Part of: 2 Anatomical structures in glaucoma > 2.5 Meshwork)
