重组人kringle 5蛋白滴眼液抑制兔角膜新生血管的研究

目的　观察重组人kringle 5 (K 5蛋白 )滴眼液抑制碱烧伤诱导的兔角膜新生血管(neovascularization ,NV)的效果 ,探讨其作用机制。方法　采用基因重组方法获得K 5蛋白 ,取新西兰白兔 4 0只 ,将浸有 1mol/L氢氧化钠滤纸片置于兔角膜中央 ,制成碱烧伤模型 ,随机均分为A、B、C及D组 ,每组 10只兔 (10只眼 ) ,伤后即分别滴用 5、10及 2 0mg/L的K 5蛋白滴眼液 ,对照 (D)组滴载体溶液 ;均每日 4次 ,共滴 2 8d。裂隙灯显微镜下观察角膜NV生长情况 ,并计算其面积 ,伤后 2 8d处死实验兔 (A、B及C组 ) ,取其角膜片做组织病理学检查。结果　伤后A、B、C及D组角膜NV开始出现的时间分别为 (3 4± 0 5 )d、(6 8± 0 4 )d、(6 7± 0 7)d及 (3 7± 0 5 )d ,其中B组明显较D组延长(P <0 0 5 ) ,而A组与D组间差异无显著意义 (P >0 0 5 )。伤后各时间点B及C组角膜NV的生长面积均明显较D组减少 (P <0 0 5 ) ,B与C组角膜NV区面积间比较 ,差异无显著意义 (P >0 0 5 )。角膜组织病理学检查 ,D组烧伤区可见大量炎性细胞浸润及角膜NV形成 ,而B及C组角膜炎性反应较轻 ,烧伤区无NV形成。角膜NV区面积与角膜后膜和炎性细胞数均有明显相关性 (P <0 0 5 )。结论重组人K 5蛋白滴眼液可明显抑制碱烧伤引起的角膜NV的生长 ,抑制

An experimental study of anti-angiogenesis with recombinant human kringle 5

OBJECTIVE: To investigate the anti-angiogenesis effects of recombinant human kringle 5 (rhk5) eye drops on corneal neovascularization (CNV) in rabbits induced by alkali burn. METHODS: Forty New Zealand albino rabbits were burned on the central corneas of their right eyes by 1 mol/L NaOH for 60s. Animals were randomly divided into 4 groups, 10 rabbits each group. Then rhk5 eye drops with different concentrations were applied four times daily for four weeks on each group: 5 mg/L in group 1, 10 mg/L in group 2, 20 mg/L in group 3 and carrier solution in group 4 (controls). The occurrence and development of CNV was observed every other day by slit-lamp microscope, and the area of CNV was calculated. Then the rabbits were killed on the 28th day (d), and the corneas were taken for histopathological examinations. Pearson's analysis was used to analyze the correlation between the number of inflammatory cells and the area of CNV. RESULTS: The occurrence of CNV in group 1 was (3.4 +/- 0.5) d, group 2 (6.8 +/- 0.4) d, group 3 (6.7 +/- 0.7) d and group 4 (3.7 +/- 0.5) d. Significant difference (P < 0.05) in the occurrence time of CNV was found between group 2 and the control. Compared with the control group, the CNV occurrence was significantly delayed in the group treated by 10 mg/L rhk5. No significant difference was found between control group and group 1, and between group 2 and group 3. The CNV areas of group 2 and group 3 were also smaller than the control group (P < 0.05). The area of CNV had close relationship with the retrocorneal membrane and with the number of inflammatory cells (P < 0.05). CONCLUSION: Topical application of recombinant human rhk5 has prominent effectiveness on the inhibition of angiogenesis induced by alkali burn.