脑出血大鼠脑组织ROCK2与脑水肿、炎性损伤及细胞凋亡的关系

Relationship between ROCK2 and cerebral edema,inflammatory injury and apoptosis in rats after intracerebral hemorrhage

目的 观察脑出血(ICH)大鼠脑水肿、炎性损伤及细胞凋亡的变化,探讨Rho相关卷曲螺旋形成蛋白激酶2(ROCK2)是否与ICH后脑损伤相关。方法 60只大鼠采用自体不凝血注入法制备ICH模型,随机分为6组:假手术组、ICH12h组、ICH1d组、ICH3d组、ICH5d组、ICH7d组。分别于相应时间点断头取脑,苏木精-伊红染色观察血肿形成及血肿周围组织炎性细胞浸润情况,并用干湿重法检测脑组织含水量,采用ELISA法检测脑组织IL-6、TNF-α水平,Westernblot法检测血肿周围组织ROCK2、Bcl-2、Bax、Caspase-3蛋白表达水平。结果 大鼠ICH后12h、1d、3d血肿周围组织ROCK2蛋白表达与假手术组相比明显增高,差异有统计学意义(P<0.05)。大鼠ICH后12h、1d、3d、5d、7d脑组织含水量、IL-6、TNF-α相对含量明显增高,与假手术组相比差异有统计学意义(P<0.05)。大鼠ICH后12h、1d、3d、5d、7d血肿周围组织Bcl-2、Bax、Caspase-3蛋白表达与假手术组相比明显增高,差异有统计学意义(P<0.05)。结论 ROCK2可能参与了ICH后脑水肿、炎性损伤及细胞凋亡等病理过程,可能为ICH的治疗提供新的靶点。

Objective To observe the changes of cerebral edema, inflammatory injury and apoptosis in rats with intracerebral hemorrhage(ICH), and to explore whether Rho associated coiled-coil forming protein kinase 2 (ROCK2) is associated with brain injury after ICH. Method Sixty rats were randomly divided into 6 groups: sham-operated group, ICH12 h, 1 d, 3 d, 5 d, 7 d group. The brain of rats was taken out at corresponding time point respectively, the formation of hematoma and the infiltration of inflammatory cells in the brain tissue were observed by hematoxylin-eosin( HE) staining. The water content of brain tissue was measured by the wet-dry method. The level of IL-6, TNF-α in brain tissue was assay by ELISA ,The expression of ROCK2, Bcl-2, Bax, Caspase-3 was detected by Western-blot. Results The expression of ROCK2 in the perihematomal tissue around the 12h, the 1d and the 3d after ICH in rats was significantly higher than that in the sham operation group( P<0.05). The content of water, the relative content of IL-6 and TNF-α in cerebral tissue of rats after 12h, 1d, 3d, 5d and 7d was significantly higher than that of sham operation group( P<0.05). The expression of Bcl-2, Bax and Caspase-3 in the perihematomal brain tissue at 12h, 1d, 3d, 5d and 7d after intracerebral hemorrhage in rats was significantly higher than that in the shamoperated group. The difference was statistically significant( P<0.05). Conclusion ROCK2 may be involved in cerebral edema, inflammatory injury and apoptosis after ICH and may provide a new therapeutic target for the treatment of ICH.