摘要
目的:观察补阳还五汤预处理对小鼠局灶性脑缺血后半暗带细胞自噬水平的影响,进一步探讨其神经保护的作用机制。方法:将80只C57BL/6小鼠随机分为假手术组、模型组、补阳还五汤组、补阳还五汤+Rapamycin(Rapa)组、Rapa组及5%DMSO组,每组12只,分别给以Rapamycin腹腔注射诱导细胞自噬及补阳还五汤灌胃给药,预处理7d后采用大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)法复制小鼠脑缺血模型,造模后3h运用2,3,5-三苯基氯化四氮唑(TTC)染色测定脑梗死体积,计算相对梗死体积,HE染色观察半暗带组织病理形态变化,Western blot法检测自噬标记性蛋白LC3-II/LC3-I比值及P62蛋白表达。结果:与假手术组比较,模型组相对梗死体积增加(P<0.05),半暗带组织形态出现缺血后病理改变,LC3-II/LC3-I比值增加(P<0.05),P62蛋白则表达减少(P<0.05);与模型组比较,补阳还五汤组相对梗死体积减少(P<0.05),半暗带组织病理形态明显改善,LC3-II/LC3-I比值减少(P<0.05),同时P62表达增加(P<0.05),而Rapa组则与补阳还五汤组相反(均P<0.05),5%DMSO组无显著差异(P均>0.05);与补阳还五汤组比较,补阳还五汤+Rapa组相对梗死体积增加(P<0.05),半暗带组织病理形态恶化,LC3-II/LC3-I比值增加(P<0.05),同时P62表达减少(P<0.05)。结论:补阳还五汤可能通过抑制脑缺血后半暗带自噬水平,减轻局灶性脑缺血小鼠脑组织损伤,发挥神经保护作用。
Objective： To investigate the effect of Buyang Huanwu decoction on autophagy in the penumbra in mice with focal cerebral ischemia and the mechanism of its neuroprotective effect. Methods： A total of 80 C57 BL /6mice were randomly divided into sham- operation group,model group,Buyang Huanwu decoction group,Buyang Huanwu decoction ＋ Rapamycin（ Rapa） group,Rapa group,and 5% dimethylsulfoxide（ DMSO） group,with 12 mice in each group. Intraperitoneal injection of Rapa was performed to induce autophagy,and Buyang Huanwu decoction was given by gavage. After 7 days of pretreatment,middle cerebral artery occlusion was used to establish a mouse model of cerebral ischemia; at 3 hours after model establishment,2,3,5- triphenyltetrazolium chloride staining was performed to measure cerebral infarct volume,and the relative infarct volume was calculated. HE staining was used to observe the histopathological changes in the penumbra,and Western blot was used to measure LC3- II / LC3- I ratio and the expression of P62 protein. Results： Compared with the sham- operation group,the model group had a significant increase in relative infarct volume（ P 〈0. 05）,pathological changes after ischemia in the penumbra,a significant increase in LC3- II / LC3- I ratio（ P 〈0. 05）,and a significant reduction in the expression of P62 protein（ P 〈0. 05）. Compared with the model group,the Buyang Huanwu decoction group had a significant reduction in relative infarct volume（ P 〈0. 05）,marked improvement in the histopathology of the penumbra,a significant reduction in LC3- II / LC3- I ratio（ P 〈0. 05）,and a significant increase in the expression of P62 protein（ P 〈0. 05）,while the Rapa group had opposite changes as the Buyang Huanwu decoction group（ all P 〈0. 05）,and there were no significant differences between the Buyang Huanwu decoction group and the 5% DMSO group（ all P 〈0. 05）. Compared with the Buyang Huanwu decoction group,the Buyang Huanwu decoction ＋ Rapa group had a significant increase in relative infarct volume（ P 〈0. 05）,deterioration in the histopathology of the penumbra,a significant increase in LC3-II / LC3- I ratio（ P 〈0. 05）,and a significant reduction in the expression of P62 protein（ P 〈0. 05）. Conclusion：Buyang Huanwu decoction can alleviate cerebral injury in mice with focal cerebral ischemia and exert its neuroprotective effect,possibly by inhibiting autophagy in the penumbra after ischemia.
出处
《湖南中医杂志》
2017年第2期119-123,共5页
Hunan Journal of Traditional Chinese Medicine
基金
国家自然科学基金资助项目(编号:81273989
81302953)
湖南省教育厅创新平台项目(编号:12K089)
湖南省益阳市科技厅一般项目(编号:YK1541)
湖南中医药大学研究生科研创新项目(编号:2015CX03)
关键词
补阳还五汤
局灶性脑缺血
细胞自噬
小鼠
实验研究
Buyang Huanwu decoction
focal cerebral ischemia
autophagy
mouse
experimental study
