原花青素对实验性脑出血大鼠脑组织细胞凋亡及凋亡相关蛋白表达的影响

目的探讨原花青素对脑出血大鼠脑组织细胞凋亡的影响及其可能机制。方法采用Ⅶ型胶原酶诱导大鼠脑出血,建立脑出血模型。54只健康SD大鼠随机分为假手术组、模型组、原花青素低剂量组(50 mg.kg-1)、中剂量组(100 mg.kg-1)、高剂量组(200 mg.kg-1)及尼莫地平对照组(10 mg.kg-1)。术后24 h,检测大鼠脑组织谷胱甘肽(GSH)含量与过氧化氢酶(CAT)的活性,采用免疫组化法检测各组大鼠脑组织切片bax和bcl-2蛋白表达,TUNEL法检测各组大鼠脑组织凋亡细胞的数量。结果与模型组比较,原花青素低、中、高剂量组大鼠脑中GSH含量与CAT活性明显升高(P<0.01)。免疫组织化学结果显示,与模型组相比,原花青素低、中、高剂量组的大鼠脑组织bax蛋白表达降低,原花青素中、高剂量组大鼠脑组织bcl-2蛋白表达升高,差异均有显著性(P<0.05,P<0.01)。TUNEL结果显示,与模型组相比,原花青素低、中、高剂量组细胞凋亡数有所减轻(P<0.05,P<0.01)。结论原花青素能够减轻脑出血大鼠的细胞凋亡,其机制可能与原花青素能够提高机体的抗氧化能力,同时增强抗凋亡基因bcl-2的表达,降低促凋亡基因bax的表达有关。

Effects of procyanidins on apoptosis and the expression of apoptosis-related proteins in brain of experimental rats with intracerebral hemorrhage

Aim To investigate the effect of procyanidins on apoptosis in brain of experimental rats with intracerebral hemorrhage(ICH) and its possible mechanisms.Method Rat intracerebral hemorrhage model was established by injecting collagenase into the brain caudate nucleus.Fifty-four health Sprague-Dawley rats were divided into six groups randomly: sham operation group,model group of intracerebral hemorrhage,treatment groups with procyanidins at doses of 50 mg·kg~-1,100 mg·kg~-1 and 200 mg·kg~-1,and positive control group with nimodipine at dose of 10 mg·kg~-1.Twentyfour hours after operation,the content of glutathione(GSH) and activity of catalase(CAT) were measured.The levels of bax and bcl-2 were detected by method of immunohistochemistry,and the apoptosis in brain tissues was detected by method of TUNEL.Result In procyanidins groups(50,100 and 200 mg·kg~-1),the content of GSH and the activity of CAT were significantly higher than those in ICH group(P<0.01).Meanwhile,procyanidins(50,100 and 200 mg·kg~-1) degraded the expression of bax in brain tissues,and procyanidins(100,200 mg·kg~-1) enhanced the expression of bcl-2(P<0.05,P<0.01).In addition,the quantity of positive TUNEL cells in procyanidins groups(50,100 and 200 mg·kg~-1) was lower than that in ICH group(P<0.05,P<0.01). Conclusions Procyanidins can decrease brain apoptosis in rats with cerebral hemorrhage which may result from raising personal ability of anti-oxidation.The modulation of bcl-2 and bax may also contribute to the mechanisms.