The Impact Of Integrin Alpha9Beta1on The Corneal Lymphangiogenesis And The Vascular Endothelial Growth Factor C

ObjectiveFor new lymphatic play an important role in the occurrence anddevelopment of inflammatory lesions in the cornea. This study make theconstruction of corneal suture inflammation model of mice, and inducedcorneal new lymphatic. To research the impact and clinlical meaning of theexogenous integrin α9β₁and its antibody Y9A2on the corneallymphangiogenesis and expression of vascular endothelial growth factor C（VEGF-C） after cornea suture．Methods80Female health BALB/c mice were divided into4groups, includenormal control group, suture control group, α9β₁group, α9β₁suppressiveedgroup,20mice in each group. normal control group without treatment. Themice of suture control group, α9β₁group, α9β₁suppressiveed group wereestablished suture model and were treated respectively with Levofloxacin/Levofloxacin+α9β₁/Levofloxacin+anti-α9β₁monoclonal antibody Y9A2. The change of the mice cornea and the growth of angiogenesis areobserved in the microscop every day. And a random sample of4mice arekilled from each group on3、5、7、14、21days after suture. Their corneaswere used for detection after death. The left corneas of two mice were usedfor examined by biopsy immunohistochemical staining for lymphaticendothelial cells. To observe lymphangiogenesis at different points andhave lymphatic vessels counting; The right corneas of this two mice wereused for immunofluorescence of lymphatic endothelial cells to observe thecornea lymphatic dyeing form.The left corneas of the other two mice used for RT-PCR to detectVEGF-C mRNA expression; Their right corneas were used to detect theVEGF-C protein expression by Western blotting method in different timepoints. Finally, the data were analysed by statistical software SPSS13.0.The data were described as x±s, by Levene inspection variance together.compare test indexes’ mean overal of each group by the single factoranalysis of variance, compares the mean between two groups by SNK-qinspection.ResultsAfter suture, observe the outlines of cornea and angiogenesis: insurgical microscope, normal mice corneas are transparent. There are arteriesnets in corneal margin, but no blood vessels in the cornea. brush shape angiogenesis were found in corneal margin of suture control group on the3nd postoperative day. The angiogenesis invaded into the cornea on5thpostoperative day. They arrived at the suture position on7thday, andInterweaves mesh. The vessels invaded into the center of cornea on14thday.Then turn back and around the suture on the21th day. compared withsuture control group, there is no significant difference of angiogenesisinα9β₁group and α9β₁suppressiveed group.no lymphatic vessels were found in the normal cornea.lymphangiogenesis were found in comeal limbus of three suture groups, andlymphatic vessels counting （LVC）reached peak level on the14thpostoperative day. lymphangiogenesis were found in comeal limbus ofsuture control group on the3rd postoperative day, the LVC is3.27±0.25.compared with suture control group, the LVC of α9β₁group was obviouslyhigher（4.93±0.38at14th day，P<0.05）, and the LVC of α9β₁suppressiveed group was noticeably lower than that in suture control grouppostoperation（2.25±0.34at14th day，P<0.05）.The VEGF-C expression is a little in normal cornea. The expression ofVEGF in suture control group was higher than normal control group andreached the peak level on14th day （P<0.05）. Compared with suture controlgroup, the expression of VEGF-C increased significantly in α9β₁groupwhile it is decreased obviously in α9β₁suppressiveed group（P<0.05） ConclusionsMice corneal suture model can induced angiogenesis and the formationof new lymphatic; Integrin α9β₁can promote the growth oflymphangiogenesis. The expression of VEGF-C and cornealymphangiogenesis are reduced by Suppressing α9β₁with antibody Y9A2.