脑出血模型大鼠c-Jun氨基末端激酶变化及阿加曲班对其表达的影响

Changes of c-Jun N-terminal kinase in rat models of intracerebral hemorrhage and effect of argathoban on it

目的 探讨大鼠脑出血后血肿周围脑组织c-Jun氨基末端激酶（JNK）表达的变化,以及凝血酶抑制剂阿加曲班对JNK活化的影响. 方法 第一部分：30只雄性SD大鼠按随机数字表法分为假手术组（6只,纹状体注射生理盐水50μL）和脑出血组（24只,纹状体注射自体血50 μL）,脑出血组再按照不同取材时间点分为12h、1d、3d、7d共4个亚组（每亚组6只）;应用Westernblotting检测各组各时间点磷酸化JNK（p-JNK）和JNK的表达变化.第二部分：36只雄性SD大鼠按随机数字表法分为假手术组（12只）、脑出血＋生理盐水组（12只,造模后3h原位注射生理盐水20μL）和脑出血＋阿加曲班组（12只,造模后3h原位注射阿加曲班20 μL）;应用Western blotting和免疫组化染色检测各组造模后1d血肿周围脑组织p-JNK和JNK的表达. 结果 （1）与假手术组比较,脑出血组各亚组血肿周围脑组织p-JNK蛋白表达均明显增高,并在脑出血后1d达到高峰,差异有统计学意义（P＜0.05）,而JNK蛋白表达无明显变化.（2）造模后1d,脑出血＋阿加曲班组血肿周围脑组织p-JNK蛋白表达较脑出血＋生理盐水组明显降低,差异有统计学意义（P＜0.05）,而JNK蛋白表达无明显变化;免疫组化染色检测到的p-JNK蛋白表达与Western blotting结果相同. 结论 脑出血可诱导血肿周围脑组织JNK活化,而阿加曲班可抑制这一过程.

Objective To study the expression of c-Jun N-terminal kinase （JNK） in the hematoma peripheral region of rat models after intracerebral hemorrhage （ICH） and explore the effect of thrombin inhibitor argathoban on JNK expression.Methods In first set,30 male SD rats weighed 250-300 g were randomized into sham-operated group （striatal injection of saline 50 μL,n=6） and ICH group （intracerebral infusion of autologous blood 50 μL at corpus striatum,n=24）; rats in ICH group were divided into four subgroups （12 h,and 1,3 and 7 d after intracerebral infusion ofautologous blood,n=6）;Western blotting was utilized to observe the changes of phosphorylated-JNK （p-JNK） and JNK at the variable groups and time points.In second set,36 male SD rats were randomized into sham-operated group （n=12）,ICH＋saline group （n=12） and ICH＋argathoban group （giving 20 μL argathoban 3 h after intracerebral infusion ofautologous blood,n=12）; Western blotting and immunohistostaining were used to observe the expressions of p-JNK and JNK in sham-operated group,ICH＋saline group and ICH＋argathoban group one day after intracerebral infusion ofautologous blood.Results （1） Western blotting indicated that the expression of p-JNK in the peripheral region of hematoma in ICH group was increased and reached its peak level one day after infusion,which was significantly higher than that in the sham-operated group （P〈0.05）; while the expression level of JNK was not altered at different groups.（2）Western blotting indicated that the expression of p-JNK in the peripheral region of hematoma in ICH＋argathoban group was statistically decreased as compared with that in the ICH＋saline group one day after infusion （P〈0.05）,while the expression level of JNK was not altered at different groups.These findings were consistent with the results of immunohistostaining.Conclusion ICH induces JNK activation in the peripheral region of hematoma and thrombin inhibitor argathoban can inhibit this process.