Dose-dependent in vitro inhibition of rabbit corneal matrix metalloproteinases by an extract of Pothomorphe umbellata after alkali injury.

The in vitro ability of Pothomorphe umbellata ethanolic crude extract to inhibit matrix metalloproteinase (MMP) in normal cornea and in cornea after alkali injury was demonstrated. Corneas of albino rabbits were injured with 1 N NaOH for 20 s. After 48 h the corneas were excised, homogenized and analyzed for MMP-9 (92 kDa), pro-MMP-2 (72 kDa) and MMP-2 (67 kDa) activity by gelatin zymography. The activity was also measured in untreated corneas. After electrophoresis of 20 microg protein, gels were incubated with 50, 100, or 250 microg/mL lyophilized hydroethanolic (1:1) root crude extract of P. umbellata standardized for 4-nerolidylcatechol (7.09%). The activity of the enzymes was compared with that of untreated gel. At 48 h after injury, the activity of all MMPs was increased compared with untreated eyes. When the gels were incubated with P. umbellata extract the activity of MMP-2, pro-MMP-2 and MMP-9 decreased in a dose-dependent manner. MMP-9 activity decreased by approximately 50% after incubation with 50 microg/mL and was completely abolished at 100 and 250 microg/mL of the extract. After incubation with 50 microg/mL the activity of pro-MMP-2 and MMP-2 also decreased by 50%. The activity of pro-MMP-2 was almost completely abolished after incubation with 250 microg/mL of the extract. For MMP-2 the incubation with 100 or 250 microg/mL of the extract of P. umbellata promoted a 10-fold decrease in activity. In conclusion, P. umbellata root crude extract can be useful as an alternative therapy to control MMP activity after corneal injury.

Dose-dependent in vitro inhibition of rabbit corneal matrix metalloproteinases by an extract of Pothomorphe umbellata after alkali injury

The in vitro ability of Pothomorphe umbellata ethanolic crude extract to inhibit matrix metalloproteinase (MMP) in normal cornea and in cornea after alkali injury was demonstrated. Corneas of albino rabbits were injured with 1 N NaOH for 20 s. After 48 h the corneas were excised, homogenized and analyzed for MMP-9 (92 kDa), pro-MMP-2 (72 kDa) and MMP-2 (67 kDa) activity by gelatin zymography. The activity was also measured in untreated corneas. After electrophoresis of 20 æg protein, gels were incubated with 50, 100, or 250 µg/mL lyophilized hydroethanolic (1:1) root crude extract of P. umbellata standardized for 4-nerolidylcatechol (7.09 percent). The activity of the enzymes was compared with that of untreated gel. At 48 h after injury, the activity of all MMPs was increased compared with untreated eyes. When the gels were incubated with P. umbellata extract the activity of MMP-2, pro-MMP-2 and MMP-9 decreased in a dose-dependent manner. MMP-9 activity decreased by approximately 50 percent after incubation with 50 µg/mL and was completely abolished at 100 and 250 µg/mL of the extract. After incubation with 50 µg/mL the activity of pro-MMP-2 and MMP-2 also decreased by 50 percent. The activity of pro-MMP-2 was almost completely abolished after incubation with 250 µg/mL of the extract. For MMP-2 the incubation with 100 or 250 µg/mL of the extract of P. umbellata promoted a 10-fold decrease in activity. In conclusion, P. umbellata root crude extract can be useful as an alternative therapy to control MMP activity after corneal injury.

[Tear enzymes in the treatment of an experimental alkaline corneal burn with gordox]. / Fermenty slezy pri lechenii éksperimental'nogo shchelochnogo ozhoga rogovitsy gordoksom.

The study of the influence of hordox, in treatment of experimental alkaline burn of the cornea, on the activity of trypsin-like proteases, elastases, callicreine, beta-N-acetylglucosaminidase and beta-glucuronidase in a tear fluid has shown that activity of these enzymes in a tear after burn remarkably increases, especially within first 24 hours and at the end of the second week after burn. In treatment by hordox, the activity of all enzymes in the tear, except elastase, reduces as compared with untreated animals, that speaks about antiinflammatory action of the preparation. On the basis of the data obtained it is suggested that investigation of hydrolytic enzymes in a tear can serve as a criterion for aimed correction of proteolysis in inflammatory processes in the cornea.

[The use of lysosomotropic preparations in treating severe experimental chemical eye burns]. / Primenenie lizosomotropnykh preparatov v lechenii tiazhelykh khimicheskikh ozhogov v éksperimente.

The action of membranotropic preparations--mildronat and phosphaden on the course of a severe burn process of the cornea with changes in lysosomal membranes revealed in pathogenesis has been studied in 80 rabbits. As a marking lysosomal enzyme, acid phosphatase was used. Besides this, peculiarities of the clinical course of the burn process have been studied, when treated by common methods and in a complex with the mentioned preparations. The results of the study have shown expressed stabilizing action of the preparations on lysosomal membranes of the cornea in early terms of the treatment, correlative relationship between results of biochemical investigations and clinical manifestations of the action of membranotropic preparations, high effectiveness of therapeutic action of mildronat as compared with phosphaden. The results obtained can serve as a foundation for the usage of the preparations in complex treatment of patients with severe chemical burns of the eye.