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目的研究低阈值经瞳孔温热疗法(TTT)对急性高眼压大鼠视网膜神经节细胞(RGC)是否具有保护作用。设计实验研究。研究对象BN大鼠。方法采用810nm二极管激光机对10只大鼠视网膜进行热刺激,照射光斑1．2mm,能量50mW,照射时间20s,干预后3d光镜下观察视网膜形态结构的改变,免疫组化方法检测HSP70、HSP27在视网膜组织表达。采用上述激光参数,照射视网膜后3d,制作急性高眼压模型(TTT+I/R组,n=10),采用TUNEL法检测RGC层细胞凋亡数量,及计数高倍镜下RGC层细胞数,与未干预的急性高眼压模型组(I/R组,n=10)、单纯TTT干预组(TTT组)及正常对照组(n=6)进行比较。主要指标免疫组化染色RGC细胞数及RGC层细胞凋亡数。结果采用低阈值TTT可诱导BN大鼠视网膜神经节细胞HSP70及HSP27表达,且光镜下未出现明显视网膜脉络膜形态的改变。TTT+I/R组RGC层细胞凋亡数量明显少于I/R组(P=0．048),且前者RGC层细胞数量明显多于后者(P=0．016);TTT组与正常对照组比较RGC层细胞凋亡数量无显著性差异(P=0．882),但RGC层细胞数明显少于正常对照组(P=0．001)。结论低阈值TTT可诱导BN大鼠视网膜HSP70、HSP27表达,并在急性高眼压损伤下对大鼠RGC凋亡具有抑制作用。(眼科,2007,16:48-51)
Objective To investigate whether low threshold transpupillary thermotherapy (TTT) has a protective effect on retinal ganglion cells (RGC) in rats with acute ocular hypertension. Design experiment research. Study object BN rats. Methods The retina of 10 rats was thermally stimulated by 810nm diode laser with a light spot of 1.2mm, an energy of 50mW and an irradiation time of 20s. Morphological changes of the retina were observed by light microscope after 3 days. Immunohistochemistry was used to detect the expression of HSP70, HSP27 Expression in retinal tissue. Acute intraocular hypertension (TTT + I / R group, n = 10) was made by using the laser parameters above the retina for 3 days. The number of apoptotic RGCs was detected by TUNEL method and the number of RGC layer cells under high magnification was counted. The rats were randomly divided into three groups: untreated group with high intraocular pressure (I / R group, n = 10), TTT group (TTT group) and normal control group (n = 6) The main indicators of immunohistochemical staining RGC cell number and RGC layer cell apoptosis number. Results The low threshold TTT induced the expression of HSP70 and HSP27 in retinal ganglion cells of BN rats, and no change of retinal choroid morphology was observed under light microscope. The number of apoptotic cells in RGC layer in TTT + I / R group was significantly less than that in I / R group (P = 0.048), and the number of RGC layer cells in the former group was significantly more than that in the latter (P = 0.016) There was no significant difference in the number of apoptotic RGCs in the control group (P = 0.882), but the number of RGC layer cells was significantly less than that in the normal control group (P = 0.001). Conclusion Low threshold TTT can induce the expression of HSP70 and HSP27 in the retina of BN rats and inhibit the apoptosis of rat RGCs under acute ocular hypertension. (Ophthalmology, 2007, 16: 48-51)