孕酮对急性高眼压大鼠视网膜神经损伤保护作用的研究

目的：通过建立大鼠急性高眼压模型，应用孕酮干预，观察孕酮是否对视网膜神经细胞具有保护作用。

方法：Wistar大鼠140只，随机分为正常对照组(A组)20只、高眼压对照组(B组)60只、高眼压孕酮干预组(C组)60只。应用生理盐水前房灌注的方法建立大鼠急性高眼压模型，C组在高眼压后即刻予腹腔注射孕酮4.0mg/kg，以后于6，24h以及隔日行皮下注射，B组于相同时间注射等量的生理盐水。于高眼压后1，3，7d分别获取各组大鼠的视网膜组织。HE染色观察视网膜病理学的变化，用免疫组化染色、Western blot检测视网膜细胞凋亡基因Caspase-3蛋白的表达情况，TUNEL法检测细胞的凋亡程度，并对各组数据进行统计学分析。

结果：B组和C组的大鼠视网膜均发生水肿， C组水肿程度明显低于B组。免疫组化染色及Western blot相对定量检测，C组Caspase-3表达程度低于B组。Caspase-3蛋白主要表达于视网膜神经节细胞层和内核层，于高眼压后第1d强度最大，以后随时间的推移逐渐减少。TUNEL检测细胞凋亡，B组和C组均出现阳性细胞，主要位于视网膜的节细胞层和内核层，C组细胞凋亡数量明显低于B组。

结论：孕酮对大鼠视网膜急性高眼压损伤有保护作用。

Protective effects of progesterone on retinal nerve injury in acute high intraocular pressure rats

AIM:To observe whether progesterone has a protective effect on retinal neural cells by establishing the acute high intraocular pressure model and intervening with progesterone.

METHODS: Totally 140 Wistar rats were randomly divided into normal control group(group A)20, high intraocular pressure control group(group B)60 and high intraocular pressure progesterone treatment group(group C)60. The acute high intraocular pressure models were made by saline anterior chamber perfusion. Group C were treated with intraperitoneal injection of progesterone 4.0mg/Kg immediately after intraocular pressure restored, and after 6, 24h and every other day were treated with subcutaneous injection. Group B were injected with equivalent saline at the same time. At the 1, 3, 7d after ocular hypertension, the retinas of rats in each group were obtained respectively. Retinas pathological changes were observed in samples hemetoxylin and eosin staining. The protein expression of Caspase-3 was measured with immunohistochemistry and Western blot. Apoptosis in retinal nerve cells were tested with TUNEL technique. Statistical analysis was performed in each group.

RESULTS: The rats retina of group B and group C were edema, but the degree in group C was lower than that in group B. In relative quantitative detection of immunohistochemistry and Western blot, expression of Caspase-3 was lower than group B. Substantial expression of Caspase-3 protein lay in cell nucleus and cytoplasm. The level of Caspase-3 in group B was higher than in group C. The positive cells were mainly in inner nucleus layer and ganglion cell layer. The level on the first day after high intraocular pressure was the highest, then gradually decreased along with the passage of time. Apoptosis tested with TUNEL technique, positive cells could be observed in group B and group C, which in inner nucleus layer and ganglion cell layer. But the number in group B was significantly more than group C.

CONCLUSION: Progesterone has nerve protection functions on retinal acute high intraocular pressure injury in rats.