Elevated intraocular pressure increases melatonin levels in the aqueous humour.

PURPOSE: To study the levels of melatonin in the aqueous humour of normotensive and hypertensive intraocular pressure (IOP) patients and to compare them to an animal model of glaucoma. METHODS: A total of 37 eyes of 37 patients who underwent cataract surgery were included in the study and were divided into normotensive patients, with IOP below 21 mmHg (n = 23), and hypertensive patients, with IOP > 21 mmHg (n = 14). Glaucomatous DBA/2J (n = 6) and control C57BL/6J (n = 6) mice presenting 3 and 12 months of age for each strain were also used. Human and mice aqueous humours were aspirated using a 30-gauge Rycroft cannula on a tuberculin syringe and further processed to quantify melatonin by high-performance liquid chromatography analysis. RESULTS: Melatonin levels in normotensive patients (IOP below 21 mmHg) presented values as medians (first quartile; third quartile) of 14.62 (5.38;37.99) ng/ml (n = 23), while hypertensive patients (IOP above 21 mmHg) showed melatonin concentrations of 46.63 (10.28; 167.28) ng/ml (n = 14; p < 0.039). Glaucoma mice presented melatonin values of 0.37 (0.34; 0.59) ng/ml (at 3 months of age, before the pathology starts), which increased to 1.55 (0.94; 1.88) ng/ml (at 12 months of age, when the pathology is fully developed and IOP is maximum; n = 6, p < 0.001). Control mice did not significantly modified melatonin concentrations between 3 and 12 months of age. CONCLUSION: Patients with high IOP present increased concentrations of melatonin in their aqueous humour compared to normotensive patients. This has been confirmed in a glaucomatous animal model in which it has been possible to see a correlation between the development of the pathology, with an increase in IOP, and a concomitant elevation of melatonin in the aqueous humour.

Silencing of P2Y(2) receptors reduces intraocular pressure in New Zealand rabbits.

BACKGROUND AND PURPOSE: P2 receptors are involved in the regulation of ocular physiological processes like intraocular pressure (IOP). In the present study, the involvement of P2Y(2) receptors in the hypertensive effect of nucleotides was investigated by use of antagonists and of a siRNA designed for the P2Y(2) receptor. EXPERIMENTAL APPROACH: Agonists of the P2Y(2) receptor a as well as P2 antagonists were applied to eyes of New Zealand rabbits, and the changes in IOP were followed for up to 6 h. Cloning of the P2Y(2) receptor cDNA was done using a combination of degenerate reverse transcription PCR (RT-PCR) and rapid amplification of cDNA ends (RACE). siRNA was synthesized and tested by immunohistochemistry. KEY RESULTS: Single doses of 2-thioUTP, UTP-γ-S and UTP increased IOP. This behaviour was concentration-dependent and partially antagonized by reactive blue 2. Silencing the P2Y(2) receptor was observed in the ciliary body by immunohistochemistry labelling, where a reduction in the immunofluorescence was observed. This reduction in the expression of the P2Y(2) receptor was concomitant with a reduction in IOP, which was measurable 24 h after treatment with the siRNA, maximal after 2 days, followed by a slow increase towards control values for the following 5 days. Application of the P2Y(2) agonists after pretreatment of the animals with this siRNA did not produce any change in IOP. CONCLUSIONS AND IMPLICATIONS: P2Y(2) receptors increase IOP in New Zealand rabbits. The application of a siRNA for this receptor significantly reduced IOP, suggesting that this technology might be used for the treatment of glaucoma.