| Objective: The aim of this study was to investigate the alteration ofAQP4 and AQP9 together with their mRNA expression in the rat eyesunder the different acute ocular hypertension (AOH), so as to explore themechanism of some diseases related to the abnormal intraocular pressure. Methods: Heathy Wistar rats (n=105, BW=250~300g) were dividedinto 2 groups randomly, the normal group(n=15) and the experimentalgroup (n=90), in which animal model of AOH was established byenhancing perfusion of physiological saline into the anterior chamber of rateye. According to the differences of perfusing pressure presented by theheight of water column (WC) from the rat eye to the surface of fluid in thebottle, the experimental rats were divided into three groups, including75cm WC ,125cm WC and 150cm WC ones. After perfusing stopped , eachgroup was subdivided into 6 subgroups at an interval of 3 h, 6 h, 12 h, 24 h ,48 h and 72 h(n=5 in each subgroup). The rats were anaesthetized with3.5% chloral hydrate(350mg/kg) and perfused with 0.9% saline and4%PFA through the ascending aorta, then the eyes were removed, slicedand stained with HE to study the pathological changes of the eye. The IHCand ISH methods were used to study AQP4, 9 and their mRNA expression.After that the imaging analysis of the AQP4, 9 and their mRNA expressionwere made by means of Computer –aided Image Analyzer, and thethickness of retina inner layer in the rats was also measured. Results: The thickness of retina inner layer in the rats was increasedat 3h after enhancing perfusion stopped , and then reached its peak at 12h.HE staining showed that the retina tissue was swelling under AOH. Theintracellular space became larger and the cells density was decreased. Theexpression of AQP4 and AQP9 in the eye tissue after AOH was strongerthan that in the control group. The AQP4 and AQP9 positive cells werelocated in the ciliary epithelium, iris epithelium, epithelium of lens , alllayers of retina and other eye areas. The protein expression reached its peakat 24h. The distribution of ISH signal was as same as that of IHC. Theexpression of AQP4 and AQP9 was significant positive correlated with itsmRNA during the retinal edema formation. The expression of AQP4 andAQP9 was correlated with the thickness of retina inner layer under AOH. Conclusions: The thickness of retina inner layer in the rats wasgradually improved and the cells in retina tissue were swelling under AOH,indicating that the retina edema appeared. The expressions of AQP4,AQP9and their mRNA up-regulated in eye tissue along with the increasing of IOP.The thickness of retina inner layer and the expression of AQP4 and AQP9all reached its peak at 12-24h under AOH. The results indicate that AQP4and AQP9 may play an important role in the retinal edema under AOH andact as pressure sensor and water channel in the process of watertransportation balance in the eye. Objective: The aim of this study was to investigate the alteration ofAQP4 and AQP9 together with their mRNA expression in the rat eyesunder the different aqueous humor osmotic gradient, so as to clarify theregulation mechanism of aqueous humor osmotic pressure . Methods: Healthy Wistar rats (n=108,BW=250~300g) were dividedinto 2 groups randomly, hypotonic perfusing group(n=54) and hypertonicperfusing group(n=54). Each group was subdivided into three subgroupsaccording to the different osmotic gradient of perfusing fluid. The rats wereanaesthetized with 3.5% chloral hydrate(350mg/kg) and perfused withD-hanks fluid presenting different osmotic pressure(268 mOsm/L,254mOsm/L,240mOsm/L;330mOsm/L,338mOsm/L, 350mOsm/L) for 60min. During the process, the perfusing pressure was maintained in 50cmWC. After perfusing stopped, the rats were subdivided into 6 subgroupsat an interval of 3h, 6h, 12h, 24h, 48h and 72 h(n=5 in each subgroup). Therats were anaesthetized with 3.5% chloral hydrate (350mg/kg) and perfusedwith 0.9% saline and 4%PFA through the ascending aorta, then the eyeswere removed, sliced and stained with HE to study the pa... |
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