MicroRNA-141-3p inhibits retinal neovascularization and retinal ganglion cell apoptosis in glaucoma mice through the inactivation of Docking protein 5-dependent mitogen-activated protein kinase signaling pathway.

Retinal neovascularization occurs in various ocular disorders including proliferative diabetic retinopathy and secondary neovascular glaucoma, resulting in blindness. This paper aims to investigate the effect of microRNA-141-3p (miR-141-3p) on retinal neovascularization and retinal ganglion cells (RGCs) in glaucoma mice through the Docking protein 5 (DOK5)-mediated mitogen-activated protein kinase (MAPK) signaling pathway. Chip retrieval and difference analysis were used for the potential mechanism of miR-141-3p on glaucoma. All modeled mice were transfected with different expression of mimic or inhibitor. The expressions of miR-141-3p, DOK5, and related genes and proteins of the MAPK signaling pathway were detected by Reverse transcription quantitative polymerase chain reaction and western blot analysis. Cell proliferation, lumen formation, and apoptosis in the retinal vascular epithelial cells and RGCs were detected using Matrigel angiogenesis and terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling assays. Moreover, a total of 63 and 294 differentially expressed genes were obtained in GSE2378 and GSE9944 chips, and 4 genes were within the intersection of the chips. In addition, the results showed that miR-141-3p was found to inhibit the DOK5 gene and activate the MAPK pathway. The number of RGCs, the expression of p38, extracellular-signal-regulated kinases (ERK), Jun N-terminal kinase (JNK), IGF-1, VEGF, HIF1-α, Bax, caspase-3, and the extent of p38, ERK, and JNK phosphorylated were decreased with miR-141-3p upregulation. Lastly, the results obtained showed that miR-141-3p inhibited the proliferation of retinal vascular epithelial cells and inhibited angiogenesis, as well as promoted apoptosis of RGCs. The study suggests that miR-141-3p inhibits retinal neovascularization in glaucoma mice by impeding the activation of the DOK5-mediated MAPK signaling pathway.

[Drug dynamics research of salvia miltiorrhiza in rabbit plasma and aqueous humor after intravenous injection].

OBJECTIVES: To assess the permeability of Danshensu at blood-ocular barrier and its characteristics of pharmacokinetics by respectively measuring the concentrations of Danshensu in blood plasma and aqueous humor of the rabbit with the high performance liquid chromatography(HPLC). METHODS: It was an experimental study. Seventy-two white rabbits were split into three groups: control group (6 rabbits), plasma group (6 rabbits) and aqueous humor group (60 rabbits). After 0.85% salt water (control group)or salvia miltiorrhiza (1.0 g/kg) (plasma and aqueous group) was injected into the vein of auris-edge. Samples of blood and aqueous humor were obtained for analysis . The analytical column was a BDS C18 stainless steed column(5 µm, 4.6 mm× 250 mm); Precolumn:YWGC18; the mobile phase consists of acetonitrile and 0.01 mol/L KH2PO4 (8:92, adjusted to pH = 2.8 with phosphoric acid). The UV detector was set at 279 nm; the flow rate was 1.0 ml/min;and the column temperature was ordinary temperature. RESULTS: The plasma concentration-time curves of Salvia miltiorrhiza fitted three-compartment model. t1/2 ß (elimination half time): 5.661 min; Cmax (peak concentration): 727.29 mg/L; Tmax(peak time ): 0 min. The aqueous humor concentration-time curves fitted two-compartment model. t1/2 ß: 147.663 min; Cmax: 38.62 mg/L; Tmax: 25 min. CONCLUSIONS: HPLC is a sensitive, specific and accurate method that can be used in pharmacokinetics research on ocular tissue of rabbit for Danshensu, Salvia miltiorrhiza that is dissolvable in water can pass through the blood-ocular barrier after intravenous injection with a relatively stable concentration of danshensu in aqueous humor resulting from a slow rate of removal.