乌司他丁对大鼠视网膜缺血再灌注损伤保护作用的研究
摘要
目的探讨乌司他丁对视网膜缺血再灌注(retinal-ischemia-reperfusi-on，RIR)损伤的保护作用及机制。
方法成年Wistar大鼠(雌雄不拘)随机分成正常对照组、缺血再灌注未治疗组、缺血再灌注乌司他丁治疗组。采用前房灌注液体形成14.3kPa高眼压而建立RIR模型。治疗组在手术前30min给予大鼠腹腔注射乌司他丁20000u／kg。缺血60min后恢复血流。应用光镜观察各组视网膜内层厚度以及浸润入视网膜的中性粒细胞数目、神经节细胞数变化；免疫组织化学染色法检测核转录因子NF-κB及TNF-α在各组视网膜中的表达。
结果视网膜缺血再灌注早期，视网膜内层水肿增厚，晚期视网膜神经节细胞数目减少及视神经纤维层萎缩变薄。乌司他丁治疗组再灌注早期视网膜水肿较轻，6h以后各时间段治疗组视网膜内层厚度均较未治疗组厚(p＜0.05)，神经节细胞数目多于未治疗组(p＜0.05)，而视网膜中的中性粒细胞数目少于未治疗组(p＜0.05)。核转录因子NF-κB及TNF-α均于再灌注后6h表达，24h达到高峰，以后开始下降，但各时间段治疗组表达强度均较未治疗组明显减弱(p＜0.05)。
结论乌司他丁对视网膜缺血再灌注损伤有治疗作用，抑制缺血再灌注损伤后的炎症反应和核转录因子NF-κB的表达是其可能的保护机制。
Objective To study the protective effect and mechanisms of ulinastatin(UTI) to retina ischemia-reperfusion(RIR) injure.
Methods Adult Wistar rates were randomly assigned to 3 groups: control, ischemia-reperfusion, ischemia-reperfusion+UTI group. The model of RIR was accomplished in Wistar adult rats by increasing the intraocular pressure to 14.3 Kp for lhour via cannulation into the anterior chamber. Rats were treated with 20000u/kg UTI at 30min before operation by intrapertoneal injected. Retinal histological changes were observes, the number of RGCs and the leukocyte infiltrating in retinal were counted, and image analysis system was usesd to detect the thickness of inner retinal layer. The expression of NF-κB and TNF-a was assessed by immunohistlchemistry.
Results During the early period of RIR, the inner layer of the retina swelled and thickned, while at the later stage the decrease of RGCs number and the atrophy and thinning of nerve fiber layer were the main changer. During the early period of reperfusion, the edema of retina was reduced in treatment group than the ischemia.The inner layers of retina of retina of the treated was thicker than the untreated. The number of RGCs of treatment group was more and the number of leukocyte was less than the untreated. The expression of NF-κB and TNF-a began to increase at 6th hour and reached the peak at 24th. With the reperfusion time prolonged, it began to deceased. The expression of the treated group was relatively obvious less than the untreated.
Conclusions UTI has therapeutical effect to retinal ischemia-reperfusion injure.Thepossible protective mechanism is that UTI can downregulate NF-κB expression and decrease inflammatory reaction induced by retinal ischemia-reperfusion.
方法成年Wistar大鼠(雌雄不拘)随机分成正常对照组、缺血再灌注未治疗组、缺血再灌注乌司他丁治疗组。采用前房灌注液体形成14.3kPa高眼压而建立RIR模型。治疗组在手术前30min给予大鼠腹腔注射乌司他丁20000u／kg。缺血60min后恢复血流。应用光镜观察各组视网膜内层厚度以及浸润入视网膜的中性粒细胞数目、神经节细胞数变化；免疫组织化学染色法检测核转录因子NF-κB及TNF-α在各组视网膜中的表达。
结果视网膜缺血再灌注早期，视网膜内层水肿增厚，晚期视网膜神经节细胞数目减少及视神经纤维层萎缩变薄。乌司他丁治疗组再灌注早期视网膜水肿较轻，6h以后各时间段治疗组视网膜内层厚度均较未治疗组厚(p＜0.05)，神经节细胞数目多于未治疗组(p＜0.05)，而视网膜中的中性粒细胞数目少于未治疗组(p＜0.05)。核转录因子NF-κB及TNF-α均于再灌注后6h表达，24h达到高峰，以后开始下降，但各时间段治疗组表达强度均较未治疗组明显减弱(p＜0.05)。
结论乌司他丁对视网膜缺血再灌注损伤有治疗作用，抑制缺血再灌注损伤后的炎症反应和核转录因子NF-κB的表达是其可能的保护机制。
Objective To study the protective effect and mechanisms of ulinastatin(UTI) to retina ischemia-reperfusion(RIR) injure.
Methods Adult Wistar rates were randomly assigned to 3 groups: control, ischemia-reperfusion, ischemia-reperfusion+UTI group. The model of RIR was accomplished in Wistar adult rats by increasing the intraocular pressure to 14.3 Kp for lhour via cannulation into the anterior chamber. Rats were treated with 20000u/kg UTI at 30min before operation by intrapertoneal injected. Retinal histological changes were observes, the number of RGCs and the leukocyte infiltrating in retinal were counted, and image analysis system was usesd to detect the thickness of inner retinal layer. The expression of NF-κB and TNF-a was assessed by immunohistlchemistry.
Results During the early period of RIR, the inner layer of the retina swelled and thickned, while at the later stage the decrease of RGCs number and the atrophy and thinning of nerve fiber layer were the main changer. During the early period of reperfusion, the edema of retina was reduced in treatment group than the ischemia.The inner layers of retina of retina of the treated was thicker than the untreated. The number of RGCs of treatment group was more and the number of leukocyte was less than the untreated. The expression of NF-κB and TNF-a began to increase at 6th hour and reached the peak at 24th. With the reperfusion time prolonged, it began to deceased. The expression of the treated group was relatively obvious less than the untreated.
Conclusions UTI has therapeutical effect to retinal ischemia-reperfusion injure.Thepossible protective mechanism is that UTI can downregulate NF-κB expression and decrease inflammatory reaction induced by retinal ischemia-reperfusion.
引文
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