Study On Recombination Ribonulease Inhibitor Inhibitively Effecting On Scar Formation After Glaucoma Filtering Operation

BACKGROUNDS: Human ribonuclease inhibitor (hRI),which is inhibitor of ribonuclease A, is ubiquitous in the cytoplasm of mammal cell and inhibits neutral ribonuclease A and alkal ribonuclease A,which can combine with Human ribonuclease inhibitor equivalently. Angiogenin is a superactive angiogenin,which can promote blood vessel formation and is found recently and plays vital role in physiological and pathological neovascular formation processof ocular region. For existing the 35 per cent homology between Ang and RNase A and the very similar stereochemical structure, both of them can combine tightly with RI as ligand with Ang'S affinity 100 times more than RNase's. Accordingly it is presumed that RI , through combining with and Ang, can inhibit neovascularity, and our research has indicated that RI can inhibit the formation of blood vessel around tumor tissue.Purpose: Applying for the production methods of genetic engineering,To make fusion protein GST-hRI soluble expression dissolvably under optimizing condition. By dropping GST-hRI to hare's eyes of trabeculectomy, to confirm that hRI can restrain the formation of Neovascularity and pathological scar in wound's granulation tissue, and establish functional filtration passage, and then raise the achievement ratio of trabeculectomy of glaucoma.Methods:The revulsive concentration and temperature of revulsivum IPTG is utilized to obtain the maximum dissolvable expression of the objective fusion protein GST-hRI. After the E.coli cultured multiply is collected and crushed with hypersound, collect supernatant of centrifugalization and separate and purify with the affinitycolumn RNase A Glutathione-Sepharose 4B, and then concentrate with the withbag filter. After simulational trabeculectomy operating on hare's eyes, the experimental group is dropped hRI eyedrop on preoperative one day and postoperative every day, and is put to death on the postoperative fortingth day for obtaining tissue of operating field. By observing the type of the postoperative filtration bubble and the microvessel density of HE-dyeing tissue slice, the inhibitory action of the fusion protein GST-hRI on the formation of postopetative Neovascularity and scar in the granulation tissue is evaluated.Results:IPTG, concentration is 0.5mmol/L and temperation is 16℃,for inducing expression about 8-10 hours,The quantities of new vessels in the granulation tissue of treatment group are less than those of control group ,therefore,scar formation is less and the propotion of functional filtering bubble is more than that of control group.Conclutions:The hRI can restrain effectively the formation of Neovascularity by using dropping eyes after the filtering operation of Glaucoma .Pathological scar in the granulation tissue of the operating field,and contribute to forming functional filtration bublles, and ensure smooth hydatoid recirculation to lower ocular tension, which raises the achievement ratio of filtering operation. . The experiment can provide some data for further application of Human ribonuclease inhibitor ,which may be developed as a kind of adjuvant drug after glaucoma filtering operation...