重组粒细胞集落刺激因子动员骨髓干细胞对大鼠脑出血后脑水肿的影响

Effect of bone marrow stem cells mobilized by recombinan human granulocyte colony-stimulating factor on brain edema in rats after intracerebral hemorrhage

目的:国内外研究报道,重组粒细胞集落刺激因子可以动员骨髓干细胞向脑梗死病损部位迁徙,减轻脑缺血后的脑水肿和脑损伤,但是,国内关于该类药物对脑出血后脑水肿影响的报道鲜见。观察重组粒细胞集落刺激因子动员骨髓干细胞对大鼠脑出血后脑水肿及血肿周边区脑组织基质金属蛋白酶9表达的影响。方法:实验于2006-03/11在泸州医学院中心实验室进行。①实验材料:健康雄性SD大鼠144只,体质量(300±20)g,由泸州医学院动物科提供。实验过程中对动物处置符合动物伦理学标准。②实验方法:将大鼠随机分为假手术组、脑出血组、治疗组,48只/组。脑出血组和治疗组参照Yang的方法,采用断尾取自体血方式制备大鼠脑出血模型。假手术组用生理盐水代替自体血,其余条件与脑出血组一致。治疗组于造模1h后腹腔注射重组粒细胞集落刺激因子60μg/kg。③实验评估:检测大鼠脑组织含水量,并采用免疫组织化学法检测大鼠脑组织基质金属蛋白酶9的表达。结果:144只大鼠中共有16只大鼠脱落,其中7只大鼠术后模型评价为0级,9只死亡。均在实验过程中予以补足。①脑出血组大鼠脑组织含水量明显高于假手术组(P<0.05),脑出血组含水量在造模后48,72h最明显,且与造模后6h、12h、24h、7d相比差异显著(P<0.05)。治疗组大鼠脑组织含水量明显低于脑出血组(P<0.05)。②脑出血组大鼠脑组织基质金属蛋白酶9的表达明显高于假手术组(P<0.05),在造模后48h、72h最明显,且与6h、12h、24h、7d相比差异显著(P<0.05)。治疗组造模后各时间点基质金属蛋白酶9的表达低于脑出血组(P<0.05)。结论:重组粒细胞集落刺激因子动员骨髓干细胞能下调脑出血后血肿周边组织基质金属蛋白酶9的表达,并减轻脑水肿的程度。

AIM： Research report that bone marrow stem cells mobilized by recombinan human granulocyte colony-stimulating factor （rhG-CSF） can migrate to lesion spot of infarction, thus decrease brain edema and brain injury after cerebral ischemia. But the report about the effect of drug on brain edema after cerebral hemorrhage is rare. This study investigated the effects of bone marrow stem cells mobilized by rhG-CSF on reducing formation of brain edema and downregulation of matrix metalloproteinase （MMP）-9 in the peripheral area after intmcerebral hemorrhage （ICH） in rats. METHODS： The experiments were performed at the Central Laboratory of Luzhou Medical College from March to November in 2006. ① 144 healthy male SD rats, （300±20） g, were provided by Animal Department of Luzhou Medical College. The experimental procedures of disposing animals were accorded with ethical standards. ②Experimental rats were assigned randomly into a sham operation group, a ICH group and a treatment group, equally. According to the method of Yang, rat models of ICH were made by the method cutting off tail of rat to obtain autoblood in the ICH and treatment groups. Rats in the sham operation group received saline instead of autoblood. Rats in the treatment group were administered with rhG-CSF （60 μ g/kg） by intraperitoneal injection after 1 hour. ③The water contents and MMP-9 were measured in each group by immunohistochemical method. RESULTS： Of 144 rats, 16 rats dropped out, among which 7 rats were estimated as 0 grade and 9 rats died, and all were supplemented, ①The water contents were higher in the ICH group than in the sham operation group （P 〈 0.05）, reaching the peak at hours 48 and 72, and the difference was significant at 6, 12, 24 hours and 7 days （P 〈 0.05 ） . The water contents were lower in the treatment group than in the ICH group （P 〈 0.05）. ②The contents of MMP-9 protein were higher in the ICH group than in the sham operation group （P 〈 0.05）, reaching the peak at hours 48 and 72, and the difference was significant at 6, 12, 24 hours and 7 days （P 〈 0.05） . The contents of MMP-9 protein were significantly lower in the treatment group than in the ICH group in each time point （P 〈 0.05）. CONCLUSION： Bone marrow stem cells mobilized by rhG-CSF can reduce MMP-9 expression in the peripheral tissues of hematoma region and relieve brain edema in ICH rats.