CAJ | 학술논문

目的探讨雷帕霉素(RAPA)对大鼠角膜新生血管(CNV)的抑制作用及其机制.方法采用成组设计法,将102只健康Wistar大鼠按随机数字法分为正常对照组(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),除正常对照组外,均选用右眼制作碱烧伤模型.利用薄膜分散法制备RAPA脂质体滴眼液,以正交设计法研究制备工艺中影响脂质体制备质量的主要影响因素,筛选出较理想的处方.碱烧伤后每日裂隙灯显微镜观察眼局部情况,并详细记录碱烧伤后大鼠角膜反应情况与新生血管生长情况,采用免疫组织化学方法与半定量逆转录聚合酶链反应(RT-PCR)方法检测角膜组织缺氧诱导因子1α(HIF-1α)与血管内皮生长因子(VEGF)的表达.实验结果采用方差分析与组间q检验进行统计学分析.结果 (1)制得的RAPA脂质体为完整的类球形,平均粒径为145.2 nm,包封率为90.02%;RAPA脂质体滴眼液中RAPA含量高,粒径合适.(2)B、C、D、E组碱烧伤后14 d,CNV面积为(28.289±0.703)、(28.005±0.801)、(20.002±1.005)、(22.300±0.853)mm2(F=159.62,P<0.05);D、E组分别与B、C组相比,CNV生长迟缓、稀疏,退化较早,相应组间CNV面积分别比较差异有统计学意义(q=47.80、46.20、34.60、32.90,均P=0.00);D组与E组间比较,CNV退化更早,面积更小,差异有统计学意义(q=13.20,P=0.00).碱烧伤后,HIF-1αmRNA、VEGF mRNA、HIF-1α蛋白、VEGF蛋白表达显著增强;其中,D、E组的HIF-1α、VEGF表达被显著抑制,D组变化更明显.结论脂质体是RAPA良好的药物载体.RAPA能通过HIF-1与VEGF途径显著抑制CNV的生长,有望成为防治CNV的一种有效方法 .(中华眼科杂志,2009,45:146-152)
목적 탐토뢰파매소(RAPA)대대서각막신생혈관(CNV)적억제작용급기궤제.방법 채용성조설계법,장102지건강Wistar대서안수궤수자법분위정상대조조(A조,6지)、감소상공백대조조(B조,24지)、감소상공백지질체대조조(C조,24지)、감소상RAPA지질체치료조(D조,24지),감소상RAPA두유치료조(E조,24지),제정상대조조외,균선용우안제작감소상모형.이용박막분산법제비RAPA지질체적안액,이정교설계법연구제비공예중영향지질체제비질량적주요영향인소,사선출교이상적처방.감소상후매일렬극등현미경관찰안국부정황,병상세기록감소상후대서각막반응정황여신생혈관생장정황,채용면역조직화학방법 여반정량역전록취합매련반응(RT-PCR)방법 검측각막조직결양유도인자1α(HIF-1α)여혈관내피생장인자(VEGF)적표체.실험결과 채용방차분석여조간q검험진행통계학분석.결과 (1)제득적RAPA지질체위완정적류구형,평균립경위145.2 nm,포봉솔위90.02%;RAPA지질체적안액중RAPA함량고,립경합괄.(2)B、C、D、E조감소상후14 d,CNV면적위(28.289±0.703)、(28.005±0.801)、(20.002±1.005)、(22.300±0.853)mm2(F=159.62,P<0.05);D、E조분별여B、C조상비,CNV생장지완、희소,퇴화교조,상응조간CNV면적분별비교차이유통계학의의(q=47.80、46.20、34.60、32.90,균P=0.00);D조여E조간비교,CNV퇴화경조,면적경소,차이유통계학의의(q=13.20,P=0.00).감소상후,HIF-1αmRNA、VEGF mRNA、HIF-1α단백、VEGF단백표체현저증강;기중,D、E조적HIF-1α、VEGF표체피현저억제,D조변화경명현.결론 지질체시RAPA량호적약물재체.RAPA능통과HIF-1여VEGF도경현저억제CNV적생장,유망성위방치CNV적일충유효방법 .(중화안과잡지,2009,45:146-152)
Objective To explore the inhibiting effect of rapamycin (RAPA) on corneal neovascularization(CNV) of rats and the functional mechanism. Methods A design group was adopted. 102 Wistar rats were divided into four groups at random, including rapamycin liposome treated group(24 rats), the rapamycin solved in bean oil treated group (24 rats), blank liposome treated group (24 rats), blank treated group (24 rats) and normal control group (6 rats). All right eyes of 96 rats were induced by alkali cauterization. Rapamycin liposome were prepared by thin film hydration and the major factors were studied by the method of orthogonal design. After alkali burn, cauterized rats were observed by slitlamp biomicroscope every day. On the 1st,4th,7th,14th days after operation,the expression of HIF-1α and VEGF were examined by immunohistochemical method and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). The analysis of variance and q test groups for analysis were adopted to analyze the results. Results (1) The bodies of RAPA liposome were intact kinds of spheres, the average diameter was 145.2 nm,and the envelopment rate was 90.02%. (2) Mter the burn of 14 d, CNV area of B,C,D and E group were (28.289±0.703), (28.005±0.801), (20.002±1.005) and (22.300±0.853) mm2(F= 159.62, P<0.05). The CNV of both the rapamycin liposome treated group and the rapamycin solved in bean oil group grew slowly and smaller than that of blank liposome treated group and blank treated group (q= 47.80,46.20, 34.60, 32.90; P=0.00). While the rapamycin liposome treated group changed more obviously than the rapamycin solved in bean oil group (q=13.20, P=0.00). After alkali burn, the expression of HIF-1α and VEGF increased dramatically, meanwhile the expression of HIF-1α and VEGF were significantly decreased by RAPA. Conclusions Liposome body is an excellent medicine carrier for the RAPA. RAPA can obviously suppress the growth of CNV. The possibly mechanism is weakening VEGF expression by inhibiting the transcription factor HIF-1α. (Chin J Ophthalmol, 2009,45:146-152)