Involvement of cysteine proteases in bFGF-induced angiogenesis in guinea pig and rat cornea.
Overexpression and activation of matrix metalloprotease (MMP) have been implicated in angiogenesis. However, the involvement of cysteine proteases, such as calpains (EC 34.22.17), is obscure. Thus, the purpose of this experiment was to study the involvement of cysteine proteases in angiogenesis induced by basic fibroblast growth factor (bFGF) in guinea pig and rat corneas using cysteine protease inhibitors. Sustained-release polymers containing bFGF were implanted into guinea pig and rat corneas to induce angiogenesis. For treatment of corneal angiogenesis, polymers containing cysteine protease inhibitors, leupeptin or SJA6017, were also implanted into corneas. Using the slit lamp, the corneas were observed for nine days after polymer implantation. Soluble proteins and albumin levels were used as markers of corneal injury by angiogenesis. bFGF induced angiogenesis in guinea pig and rat corneas. In guinea pig cornea, wet weight, the amount of soluble protein and albumin was highest at four days after bFGF-containing pellet implantation. In rat cornea, the amount of soluble protein and albumin was highest at six days, and wet weight increased within four days. One hundred nmole of leupeptin showed a tendency to reduce bFGF-induced angiogenesis in guinea pig cornea, and 10 nmole of SJA6017 was effective in reducing bFGF-induced angiogenesis in rat cornea, although SJA6017 showed a stronger effect than leupeptin. Ten nmole of SJA6017 significantly reduced the number of new blood vessels. These data suggested involvement of cysteine proteases in angiogenesis in guinea pig and rat cornea.
Tamada
Y
Y
Research Laboratories, Senju Pharmaceutical Corporation, Ltd., Kobe, Hyogo, Japan.
Fukiage
C
C
Boyle
D L
DL
Azuma
M
M
Shearer
T R
TR
D000418:Albumins / Q000378:metabolism; D000818:Animals; D003315:Cornea / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D016510:Corneal Neovascularization / Q000139:chemically induced / Q000201:enzymology* / Q000473:pathology / Q000517:prevention & control; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000627:therapeutic use; D003692:Delayed-Action Preparations; D004151:Dipeptides / Q000627:therapeutic use; D005136:Eye Proteins / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000633:toxicity; D006168:Guinea Pigs; D007976:Leupeptins / Q000627:therapeutic use; D008297:Male; D009929:Organ Size; D051381:Rats
D000418:Albumins; D015853:Cysteine Proteinase Inhibitors; D003692:Delayed-Action Preparations; D004151:Dipeptides; D005136:Eye Proteins; D007976:Leupeptins; C109145:N-(4-fluorophenylsulfonyl)-L-valyl-L-leucinal; D016222:Fibroblast Growth Factor 2; D003546:Cysteine Endopeptidases; C032854:leupeptin