摘要
目的:探讨补阳还五汤对脑缺血再灌注大鼠脑组织AKT蛋白表达的影响。方法:选取SPF级SD雄性大鼠90只,将大鼠随机分为五组,假手术组10只,模型组、阳性组、高剂量补阳还五汤组、低剂量补阳还五汤组各20只。以阳性组为标准对照,模型组为空白对照。假手术组、模型组分别给10 mg/kg蒸馏水,阳性组给予6.8 mg/kg硫酸氢氯吡格雷,低剂量、高剂量组大鼠每天给予13 g/kg、26 g/kg补阳还五汤溶液,持续2周。取大脑中海马体部分固定、包埋后,行石蜡切片及HE染色。光镜下观察海马体组织病理组织学变化。制备蛋白样品、电泳、转膜、发生免疫反应,凝胶成像分析软件检测目标蛋白荧光强度,测定AKT、p-AKT蛋白表达。结果:模型组海马组织细胞排列紊乱,数量减少,细胞核固缩深染,染色质凝聚,空泡样变性坏死数量增加。假手术组海马体区不产生缺血性表现,细胞形态、结构正常。给药组多数神经细胞结构、形态改变相对较轻,核固缩少,且高剂量组对脑损伤的改善程度大于低剂量组。模型组p-AKT阳性细胞率显著高于假手术组(P<0.01);阳性组、补阳还五汤高剂量组p-AKT阳性细胞率显著高于模型组(P<0.01)。模型组p-AKT蛋白表达水平显著低于假手术组(P<0.01);阳性组、补阳还五汤高剂量组p-AKT蛋白表达水平显著高于模型组(P<0.01);各组间AKT蛋白表达无统计学差异(P>0.05)。结论:补阳还五汤可激活PI3K/AKT信号通路,信号通路中AKT、p-AKT蛋白表达升高,促进AKT磷酸化。补阳还五汤还可保护神经血管结构和功能,改善脑缺血再灌注组织病理损伤,发挥保护脑作用。
Objective： To explore effects of Buyang Huanwu decoction on AKT protein expression in brain tissue of rats with cerebral ischemia reperfusion injury. Methods： 90 SPF SD male rat were selected and divided them into five groups randomly： 10 rats in sham group,20 rats in model group（ blank control）,positive group（ standard control）,high dose Buyang Huanwu decoction group and low dose Buyang Huanwu decoction group. 10 mg/kg distilled water was given to the model group and the sham group,6.8 mg/kg of Clopidogrel sulfate was given to the positive group,and 13 g/kg and 26 g/kg Buyang Huanwu decoction was given to the high dose Buyang Huanwu decoction group and the low dose Buyang Huanwu decoction group,respectively. The administration process lasted for continuous 2 weeks. The hippocampus in the brain was collected,fixed,embedded in paraffin and HE stained. The histopathological changes of hippocampus were observed under light microscope. The protein samples were prepared for electrophoresis,transfer membrane,and immune response. The gel imaging software was used to detect the target protein fluorescence intensity,and then the AKT and p-AKT protein expressions were determined. Results： In the model group,the hippocampus cells were arranged disorderly,the number was reduced,the nucleus pyknosis occurred and the nucleus was deeply stained,chromatin was condensed,and vacuolar degeneration and necrosis increased. In the sham group,the hippocampus region did not show any ischemic manifestations; however,the cell morphology and structure were normal. In the administration group,most of the nerve cells had less structural and morphological changes,and there were less nuclear condensation. The high dose group had a greater degree of brain damage than the low-dose group.The positive rate of p-AKT in the model group was significantly higher than that in the sham group（ P〈0.01）,the positive rate of pAKT in the positive group and the high dose Buyang Huanwu decoction group was significantly higher than that in the model group（ P〈0.01）; however,there was no statistical difference in the p-AKT positive rate between the low dose Buyang Huanwu decoction group and the model group. The expression level of p-AKT protein in the model group was significantly lower than that in the sham group（ P〈0.01）,the expression levels of p-AKT protein in the positive group and the high dose Buyang Huanwu decoction group were significantly higher than those in the model group（ P〈0.01）; however,there were no statistical differenced in the p-AKT protein expression among each group（ P〈0.05）. Conclusions： Buyang Huanwu decoction can activate PI3 K/AKT signaling pathway,increase AKT and p-AKT protein expression in signal pathway and promote AKT phosphorylation. Moreover,Buyang Huanwu decoction can also protect nerve vessel structure and function,improve cerebral ischemia reperfusion tissue pathological damage,and play a protective brain effect.
作者
马杰
谢薇
MA Jie;XIE Wei(The First Affiliated Hospital of Henan University of TCM,Zhengzhou Henan 450000,Chin)
出处
《中国民康医学》
2018年第9期1-3,共3页
Medical Journal of Chinese People’s Health
关键词
补阳还五汤
脑缺血再灌注
AKT蛋白
硫酸氢氯吡格雷
Buyang Huanwu decoction
Cerebral ischemia and reperlhsion
AKT protein
Clopidogrel sulfate
