摘要
目的:观察大鼠脑内注射凝血酶后水通道蛋白4表达及脑水肿的变化以及水通道蛋白4表达与血脑屏障通透性之间的关系,分析凝血酶对水通道蛋白4表达的影响。方法:实验于2005-01/11在哈尔滨医科大学第一临床医学院中心实验室完成。取雄性Wistar大鼠90只应用数字表法随机分为6组(n=15),注凝血酶6,24,72h,5,7d组及假手术组。除假手术组外,各组大鼠脑右尾状核内注射凝血酶10U(50μL)。各组大鼠在相应时间点处死,假手术组在术后2h处死,分别进行注射点周围免疫组织化学染色和血脑屏障通透性、脑组织含水率的测定。应用伊文思蓝测定血脑屏障通透性,应用干湿重法测定脑含水率,免疫组织化学染色结果应用阳性细胞计数法评估。结果:90只大鼠全部进入结果分析。①水通道蛋白4染色阳性细胞数:注凝血酶6h组即高于假手术组[(39.53±2.07),(18.40±1.24)个/视野,P<0.01],注凝血酶24h组达到高峰,为(51.93±1.67)个/视野,随后下降,注凝血酶7d组仍高于假手术组[(21.60±1.45)个/视野,P<0.01]。②脑组织含水率:注凝血酶6h组即高于假手术组(P<0.01),注凝血酶24h组达到高峰,随后下降,注凝血酶7d组仍高于假手术组(P<0.01)。③脑组织伊文思蓝含量:注凝血酶6h组即高于假手术组(P<0.01),注凝血酶24h组达到高峰,随后下降,注凝血酶7d组仍高于假手术组(P<0.01)。结论:脑内注射凝血酶,可导致注射点周围水通道蛋白4表达的增加,其变化趋势与脑组织含水率、血脑屏障通透性的变化相一致。提示水通道蛋白4参与了大鼠脑内注射凝血酶后的脑水肿形成,其变化可能是凝血酶破坏了血脑屏障完整性的结果。
AIM： To study the effects of thrombin on the aquaperins-4 （AQP4） expression and changes of brain edema （BE） as well as the relationship between AQP4 expression and the permeability of blood brain barrier （BBB） following intracerebral injection of thrombin in rats, and analyze the effects of thrombin on AQP4.
METHODS： The experiment was conducted in the Central Laboratory of First Clinical Medical College of Harbin Medical University between January and November 2005. Ninety male Wistar rats were divided randomly into six groups （n=15）, sham-operation group, 6 hours, 24 hours, 72 hours, 5 days and 7 days after thrombin injection groups. 10 U （50μL） of thrombin was injected in the eaudate nucleus of right cerebrum in rats of all groups except the sham-operation group. Rats were executed at each time point, and rats in the sham-operation group were executed at 2 hours after operation, whose permeability of BBB and the brain water content were detected after being immunohistochemically stained in the periphery of injection point. Evans-Blue was adopted to determine the permeability of BBB and the brain water content was detected with dry-wet weight method. Results of immunohistochemical staining was evaluated by positive cytometry.
RESULTS： A total of 90 rats were involved in the analysis of results.① Compared with sham operation group, the number of AQP4 positive staining cells increased significantly in 6-hours group [（39.53±2.07）, （18.40±1 .24 ）/field of vision,P 〈 0.01],and researched the peak in the 24-hour group, which was （51.93±1.67）/field of vision, then declined, while that in the 7-day group was still higher than the sham-operation group [（21.60±1.45）/field of vision,P 〈 0.01]. ②The brain water ratio： that in the 6-hour group was higher than the sham-operation group （P 〈 0.01） and peaked in the 24-hour group, and then declined, while that in the 7-day group was still higher than the sham-operation group （P 〈 0.01）.③ Evans blue contents in brain： that in the 6-hour group was higher than the sham-operation group （P 〈 0.01）, and peaked in the 24-hour group, and then declined, while that in the 7-day group was still higher than the sham-operation group （P 〈 0.01）.
CONCLUSION： The expression of AQP4 around the injection point increases following intracerebral injection of thrombin in rats, the changing trend of which is in accordance with the brain water ratio and permeability of BBB, which indicate that AQP4 has participated in the formation of BE following intracerebral injection of thrombin in rats, and the changes may be resulted from BBB breakdown by thrombin.
出处
《中国临床康复》
CAS
CSCD
北大核心
2006年第30期86-88,共3页
Chinese Journal of Clinical Rehabilitation
