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莫索尼定局部给药后的反相高效液相色谱定量分析及视网膜神经保护作用

Quantitative analysis by reversed-phase high-performance liquid chromatography and retinal neuroprotection after topical administration of moxonidine.

作者信息

Zhang Qian, Chu Mei-Fang, Li Yan-Hong, Li Chun-Hua, Lei Run-Jia, Wang Si-Cen, Xiao Bao-Jun, Yang Jian-Gang

机构信息

Department of Ophthalmology, Xi'an Fourth Hospital, Shaanxi Ophthalmological Hospital, Xi'an 710004, Shaanxi Province, China.

Xi'an Eye Hospital, First Affiliated Hospital of Northwest University; Xi'an No.1 Hospital, Xi'an 710002, Shaanxi Province, China.

出版信息

Int J Ophthalmol. 2020 Mar 18;13(3):390-398. doi: 10.18240/ijo.2020.03.04. eCollection 2020.

Abstract

AIM

To determine moxonidine in aqueous humor and iris-ciliary body by reversed-phase high performance liquid chromatography (RP-HPLC), and to evaluate the retinal neuroprotective effect after topical administration with moxonidine in a high intraocular pressure (IOP) model.

METHODS

The eyes of albino rabbits were administered topically and ipsilaterally with 0.2% moxonidine. A RP-HPLC method was employed for the identification and quantification of moxonidine between 2 and 480min, which presented in the aqueous humor and iris-ciliary body. Flash electroretinography (F-ERG) amplitude and superoxide dismutase (SOD) level were measured between day 1 and day 15 after topical administration with moxonidine in a rabbit model of high IOP. Histological and ultrastructural observation underwent to analyze the changes of retinal morphology, the inner retinal layers (IRL) thickness, and retinal ganglion cell (RGC) counting.

RESULTS

Moxonidine was detectable between 2 and 480min after administration, and the peak concentration developed both in the two tissues at 30min, 0.51 µg/mL in aqueous humor and 1.03 µg/g in iris-ciliary body. In comparison to control, F-ERG b-wave amplitude in moxonidine eyes were significantly differences between day 3 and day 15 (<0.01) in the high IOP model; SOD levels were significantly higher at all time-points (<0.01) with a maximum level of 20.29 U/mgprot at day 15; and RGCs were significantly higher (<0.05).

CONCLUSION

Moxonidine is a viable neuroprotective agent with application to high IOP model. All layers of retina, including RGC layer, retinal nerve fiber layer and INL, are more preserved after moxonidine administration. SOD plays a neuroprotective role in ocular hypertension-mediated RGC death.

摘要

目的

采用反相高效液相色谱法(RP-HPLC)测定房水和虹膜睫状体中的莫索尼定,并评估在高眼压(IOP)模型中局部应用莫索尼定后的视网膜神经保护作用。

方法

对白化兔的眼睛进行同侧局部给予0.2%莫索尼定。采用RP-HPLC法在2至480分钟内对房水和虹膜睫状体中存在的莫索尼定进行鉴定和定量。在高眼压兔模型中局部应用莫索尼定后第1天至第15天,测量闪光视网膜电图(F-ERG)振幅和超氧化物歧化酶(SOD)水平。进行组织学和超微结构观察以分析视网膜形态变化、视网膜内层(IRL)厚度和视网膜神经节细胞(RGC)计数。

结果

给药后2至480分钟可检测到莫索尼定,30分钟时在两种组织中均出现峰值浓度,房水中为0.51μg/mL,虹膜睫状体中为1.03μg/g。与对照组相比,在高眼压模型中,莫索尼定组眼睛的F-ERG b波振幅在第3天和第15天有显著差异(<0.01);SOD水平在所有时间点均显著更高(<0.01),第15天最高水平为20.29U/mgprot;RGCs显著更高(<0.05)。

结论

莫索尼定是一种适用于高眼压模型的可行神经保护剂。应用莫索尼定后,视网膜的所有层,包括RGC层、视网膜神经纤维层和内核层,得到更好的保留。SOD在高眼压介导的RGC死亡中发挥神经保护作用。

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