氨基胍对氧诱导的视网膜病变小鼠视网膜神经细胞的保护作用

Protective effects of aminoguanidine on retinal nerve cells in mice with hyperxia-induced retinopathy

目的探讨氨基胍(aminoguanidine,AG)对氧诱导的视网膜病变小鼠视网膜神经细胞的保护作用。方法 7d龄C57BL/6J小鼠80只随机分成4组:A组(正常对照组)、B组(单纯高氧组)、C组(高氧生理盐水组)、D组(高氧AG治疗组),每组20只。B组、C组、D组乳鼠置于含体积分数75%氧的氧箱中饲养12d后放于正常空气中饲养至第17天;A组一直在正常空气中饲养。D组于第7天开始腹腔注射AG(100mg·kg-1)每天1次直至第17天,C组注射等量的生理盐水。4组分别于饲养后第14天和第17天随机处死10只取视网膜做HE染色观察细胞形态、免疫组织化学染色观察诱导型一氧化氮合酶(iN-OS)的表达情况、TUNEL染色观察细胞凋亡情况。结果 iNOS阳性细胞主要表达于视网膜内核层和神经节细胞层,且B组14d(23.00±2.07)明显强于17d(12.20±1.97),B组与A组(0.00±0.00)比较差异有统计学意义(P<0.05),D组(10.13±1.59、4.93±1.03)与B组、C组(23.00±2.20、12.00±2.23)比较差异均有统计学意义(均为P<0.05)。TUNEL染色凋亡细胞主要位于视网膜内核层,B组14d(29.40±4.25)明显多于17d(7.73±1.57);B组凋亡细胞数明显多于A组(0.00±0.00)(P<0.05),D组(10.73±1.62、4.33±1.04)明显少于B组和C组(28.66±4.38、8.13±1.30),差异均有统计学意义(均为P<0.05)。结论 AG可抑制氧诱导的视网膜病变小鼠视网膜神经细胞凋亡,并可能与iNOS有关。

Objective To explore the protective effects of aminoguanidine （AG） on retinal nerve cells in mice with hyperxia-induced retinopathy. Methods Eighty 7- day-old C57BL/6J mice were divided into four groups：group A（ normal group）, group B （high oxygen group ）, group C （ high oxygen saline group ）, group D （ AG treatment group） ,20 cases in each group. The mice in group B, C, D were put into the environ- ment exposed to 75% oxygen for 12 days and backed to room air for 17 days. Group A were breed constantly in the room air. Group D were given AG（ 100 mg · kg^-1 ） from the 7th day to the 17th day. Group C were given the same amount of saline. Ten mice in four groups were randomly sacrificed to take the retina on the 14th and 7th day. The ret- inal tissue was prepared to observe the cell morphous by HE staining,the expression of inducible nitric oxide synthase （iNOS） by immunohistochemical staining, and the cell apoptosis by TUNEL method. Results The positive iNOS cells were mainly located in the inner nuclear layer and ganglion layer,the expression on the 14th day in group B was 23.00 ± 2.07 ,higher than that on the 17th day（ 12.20 ± 1.97 ）, there was statistical difference between group B and A（P 〈 0.05 ）, and there were statistical differences between group D（ 10. 13 ± 1.59,4.93 ± 1.03 ） and group B, C （ 23.00 ± 2.20,12.00 ± 2.23 ） （both P 〈 0.05 ）. The TUNEL positive cells were mainly located in retinal inner nuclear layer,the number of apoptotic cells on the 14th day in group B was 29.40 ±4.25 ,higher than that on the 17th day（7.73 ± 1.57） ,there was statistical difference between group B and A（P 〈0.05 ） ,and there were statistical differences between group D（ 10.73 ± 1.62, 4.33 ± 1.04） and group B, C （28.66 ± 4.38,8.13 ± 1.30） （ both P 〈 0.05 ）. Conelusion AG can inhibit retinal neuronal apoptosis induced by hyperxia, and the mechanism may be related with iNOS.