雷帕霉素脂质体滴眼液制备及其对角膜新生血管作用

PREPARATION OF RAPAMYCIN LIPOSOME EYE DROPS AND ITS EFFECT ON CORNEAL NEOVASCULARIZATION

目的探讨雷帕霉素(RAPA)脂质体滴眼液的制备方法及其对大鼠角膜新生血管(CNV)的治疗作用。方法应用薄膜分散法制备RAPA脂质体滴眼液,以正交设计法研究制备工艺中脂质体制备质量的主要影响因素,筛选出较理想处方。用透射电镜观察其形态,以激光散射法测定其粒径大小。将102只健康Wistar大鼠随机分为正常对照组(A组,6只)、碱烧伤空白对照组(B组,24只)、碱烧伤空白脂质体对照组(C组,24只)、碱烧伤RAPA脂质体治疗组(D组,24只)、碱烧伤RAPA豆油治疗组(E组,24只)。A组不处理;B、C、D、E均制作右眼碱烧伤动物模型,然后D组应用RAPA脂质体滴眼液滴眼,E组用RAPA豆油滴眼液滴眼,C组用空白脂质体滴眼液滴眼,B组不予处理。术后第1天开始每日裂隙灯显微镜观察眼局部情况,记录碱烧伤后4、7、14d大鼠角膜反应与CNV生长情况。结果所制RAPA脂质体为完整的类球形,平均粒径145.2nm,包封率90.02%,可作为滴眼液用于动物实验。4组大鼠CNV面积均随时间增加而增加,差异均有显著性(F=906.43~1 682.16,P<0.01)。碱烧伤4、7、14d时,B、C组CNV面积比较差异无显著性(P>0.05);其余各组间比较,D、E组CNV面积较B、C组减小,D组面积较E组减小,差异均有显著性(F=309.37~574.92,q=11.98~47.90,P<0.01)。结论脂质体是RAPA良好的药物载体。RAPA能显著抑制CNV生长,有望成为防治CNV的一种有效方法。

Objective To explore the preparation method of rapamycin （RAPA） liposome eye drops and its effect on cor- neal neovascularization （CNV） in rats. Methods Rapamycin liposome eye drops were prepared by thin film hydration. The ma- jor factors on the preparation of RAPA liposome were studied by orthogonal design. The ideal prescription was then screened out. Using transmission electron microscope （TEM） to observe the morphology of liposome, and the particle size was measured by laser light scattering method. This study consisted of 102 Wistar rats that were divided into five groups at random as follows： group A （normal control group, n = 6）, group B （blank alkali burn control group, n = 24）, group C （blank alkali burn-blank liposome blank control group, n = 24）, group D （alkali burn treated with RAPA and liposome group, n = 24） and group E （alkali burn trea- ted with RAPA soybean oil group, n ：24）. No handle was given to the rats in group A. A model of corneal alkali burn of the right eye was created in rats of groups B, C, D and E. The eyes in the rats of group D were treated with RAPA and liposome, that in group E with RAPA soybean oil, that in group C with blank liposome, and that in group B no treatment was given. From the first day after surgery, the ocular conditions were observed using slit-lamp microscope every day, the corneal reactions and the growth of CNV on days 4,7 and 14 were recorded. Results The prepared RAPA liposomes were complete spherical with an average di- ameter of 145.2 nm,and the entrapment efficiency was 90.02~, which could he used as eye drops in animal experiment. The area of CNV of rats in the four groups increases over time, the differences were signifieant （F=906.43-1 682.16,P^0.01）. On days 4,7 and 14 of alkali burn, the area of CNV between groups 13 and C was not statistically different （P〈0.05）, between the other groups, the area of CNV in groups D and E was smaller than that in groups B and C （F=309.37-574.92,q=11.98-47.90,P〈 0,01）. Conclusion Liposome is a good drug carrier of RAPA that can obviously suppress the growth of CNV, and is expected to become an effective method in prevention and cure for corneal neovascularization.