CAJ | 학술논문

目的观察生长停滞及DNA损伤诱导基因153(GADD153)和caspase-12在脑缺血再灌注损伤大鼠脑组织中的动态改变,探讨内质网应激在脑缺血再灌注损伤中的作用. 方法 42只大鼠按随机数字表法分为正常对照组(3只)、假手术组(3只)和脑缺血再灌注损伤组(36只)；脑缺血再灌注损伤组又分为脑缺血2h再灌注6h、12h、24 h、72 h组,每组各9只.采用线栓法建立大鼠大脑中动脉缺血再灌注损伤模型.采用免疫组化染色、免疫荧光双标染色、Westem blotting检测各组大鼠脑组织中GADD153和caspase-12的表达. 结果免疫组化染色和Western blotting结果均显示正常对照组和假手术组大鼠脑组织中GADD 153和caspase-12表达为阴性；再灌注6h组GADD153表达增加,并逐渐增高,持续至72 h时较再灌注6h组明显增高,差异均有统计学意义(P＜0.05)；再灌注6h组caspase-12表达增加,至24 h达高峰,72 h时仍维持在较高水平,与再灌注6h组比较差异有统计学意义(P＜0.05).免疫荧光双标染色结果显示,再灌注6h组可见少量双标阳性细胞,12h、24 h时双标阳性细胞数均较再灌注6h组明显增多,差异有统计学意义(P＜0.05),至72 hcaspase-12单标阳性细胞数减少,GADD 153单标阳性细胞数仍较多,双标阳性细胞数减少. 结论 GADD153和caspase-12在脑缺血再灌注损伤大鼠脑组织中的表达随时间呈动态改变,表明内质网应激参与了脑缺血再灌注损伤的病理过程.
목적 관찰생장정체급DNA손상유도기인153(GADD153)화caspase-12재뇌결혈재관주손상대서뇌조직중적동태개변,탐토내질망응격재뇌결혈재관주손상중적작용. 방법 42지대서안수궤수자표법분위정상대조조(3지)、가수술조(3지)화뇌결혈재관주손상조(36지)；뇌결혈재관주손상조우분위뇌결혈2h재관주6h、12h、24 h、72 h조,매조각9지.채용선전법건립대서대뇌중동맥결혈재관주손상모형.채용면역조화염색、면역형광쌍표염색、Westem blotting검측각조대서뇌조직중GADD153화caspase-12적표체. 결과 면역조화염색화Western blotting결과균현시정상대조조화가수술조대서뇌조직중GADD 153화caspase-12표체위음성；재관주6h조GADD153표체증가,병축점증고,지속지72 h시교재관주6h조명현증고,차이균유통계학의의(P＜0.05)；재관주6h조caspase-12표체증가,지24 h체고봉,72 h시잉유지재교고수평,여재관주6h조비교차이유통계학의의(P＜0.05).면역형광쌍표염색결과현시,재관주6h조가견소량쌍표양성세포,12h、24 h시쌍표양성세포수균교재관주6h조명현증다,차이유통계학의의(P＜0.05),지72 hcaspase-12단표양성세포수감소,GADD 153단표양성세포수잉교다,쌍표양성세포수감소. 결론 GADD153화caspase-12재뇌결혈재관주손상대서뇌조직중적표체수시간정동태개변,표명내질망응격삼여료뇌결혈재관주손상적병리과정.
Objective To observe the dynamic changes of growth arrest and DNA damage inducible gene 153 (GADD 153) and caspase-12 expressions in the brain of rats after ischemia-reperfusion,and explore the role of endoplasmic reticulum stress in ischemia-reperfusion brain injury.Methods Forty-two rats were randomly divided into control group (n=3),sham-operated group (n=3) and ischemia-reperfusion group (n=36); the rats of ischemia-reperfusion group were randomly sub-divided into groups of 2 h occlusion and 6,12,24,72 h reperfusion (n=9).Modified Longa intraluminal thread method was adopted to establish the middle cerebral artery occlusion-reperfusion models.Expression changes of GADD 153 and caspase-12 at different time points were detected by immunohistochemistry,double-label immunofluorescence and Western blotting.Results Immunohistochemistry and Western blotting showed that the expressions of GADD153 and caspase-12 in the control group and sham-operated group was negative; the GADD 153 expression increased in group of 6 h reperfusion,and that in group of 72 h reperfusion was significantly higher than that in group of 6 h reperfusion (P＜0.05);the caspase-12 expression increased in group of 6 h reperfusion,enjoyed the highest level in group of 24 h reperfusion which had significant difference as compared with that in group of 6 h reperfusion (P＜0.05),and still maintained at a high level in group of 72 h repefusion.Double immunofluorescence staining showed that GADD153 and caspase-12 double-positive cells could be seen in group of 6 h reperfusion;that at groups of 12 and 24 h reperfusion significantly increased as compared with that in group of 6 h reperfusion (P＜0.05); the number of caspase-12 single-positive cells reduced,that of GADD153 single-positive cells was still large,and the number of double-positive cells reduced.Conclusion The expression changes of GADD153 and caspase-12 are time dependent,indicating that endoplasmic reticulum stress involve in the pathological process of ischemia-reperfusion brain injury.