Diquafosol Tetrasodium is a P2Y2 receptor agonist that stimulates the secretion of mucins from ocular tissues Diquafosol tetrasodium (INS365) improves tear film stability and visual function, as a topical treatment of dry eye disease.

曝露时间和diquafosol浓度显著降低了细胞存活率并增加了细胞裂解。相反，HA组中无论曝露时间长短，观察到的差异均不显著。用30%稀释的diquafosol处理1小时后以及用所有浓度稀释的diquafosol处理6小时后，细胞活力显著下降。经过24小时所有浓度稀释的diquafosol处理后，细胞存活率降低，裂解增加，观察到显著的细胞损伤[1]。

在大鼠干眼模型中，P2Y(2) 激动剂 INS365 能够改善眼表健康，这一发现基于几方面的增加：泪液分泌、角膜上皮抵抗力以及杯状细胞释放含有糖蛋白的组分。因此，INS365 是治疗干眼综合征的潜在治疗剂。

The viabilities of human corneal epithelial cells (HCECs) were determined using a MTT assay.?Cells (100 μl;?5×104 cell/mL) were plated in 96-well tissue-culture plates and incubated at 37℃ in 5% CO2 for 24 to 48 h until cultures were subconfluent.?Diquafosol (100 μl diluted 10%, 20%, or 30%) or 0.3% or 0.18% HA were added and incubated for 1, 6, or 24 h. DMEM (100 μl) was added to controls.?After 1, 6, and 24 h, plates were washed three times with PBS to remove the drugs.?Cell viabilities were evaluated after incubating for 24 h. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide;?thiazoyl blue (5 mg/mL) was then added to each well.?Samples were incubated in the dark for 4 h at 37℃, and media were then removed.?Precipitates were resuspended indimethyl sulfoxide (100 μl;?DMSO, Sigma-Aldrich).?Absorbances were measured on a plate reader at 570 nm.?The experiment was performed in triplicate[1].

An SD rat dry eye model was used in which exorbital lacrimal gland extirpation decreased the Schirmer test score by at least 50%. After 8 weeks, when significant increases occurred in corneal epithelial permeability, Diquafosol tetrasodium(INS365)-containing eye drops were applied six times daily for the next 4 weeks at concentrations from 0.03% to 3.0%. Corneal barrier function was evaluated based on measurements with a modified anterior fluorometer of fluorescein penetrance at 1, 2, and 4 weeks after initial application. After INS365 application, the periodic acid-Schiff reagent (PAS)-stained area was evaluated in histologic sections of the tarsal and bulbar conjunctiva[2].