摘要
目的探讨补阳还五汤对脑出血大鼠脑组织水通道蛋白4(AQP4)表达及血-脑脊液屏障通透性的影响。方法 2014年3月—2015年11月,采用随机数字表法将成年雄性SPF级SD大鼠分为假手术组、模型组、补阳还五汤低剂量组、补阳还五汤中剂量组、补阳还五汤高剂量组,各24只。其中模型组、补阳还五汤各剂量组制备脑出血模型。造模后第2天起,补阳还五汤低剂量组、补阳还五汤中剂量组、补阳还五汤高剂量组灌胃补阳还五汤,药量分别为13.2、26.5、53.0 g·kg^(-1)·d^(-1),连续给药14 d,假手术组及模型组给予等体积的0.9%氯化钠溶液。免疫组织化学法检测磷酸化肌醇磷脂3激酶(PI3K)、蛋白激酶B(AKT)表达水平,免疫荧光标记法检测AQP4表达水平,干湿重法检测脑组织含水量,甲酰胺法检测血-脑脊液屏障通透性〔以依文思蓝(EB)含量表示〕。结果假手术组、模型组、补阳还五汤低剂量组、补阳还五汤中剂量组、补阳还五汤高剂量组PI3K、AKT表达水平比较,差异均有统计学意义(P<0.05);其中补阳还五汤低剂量组、补阳还五汤中剂量组、补阳还五汤高剂量组PI3K表达水平高于模型组;补阳还五汤中剂量组、补阳还五汤高剂量组AKT表达水平高于模型组(P<0.05)。5组AQP4表达水平比较,差异有统计学意义(F=95.79,P<0.001);其中补阳还五汤低剂量组、补阳还五汤中剂量组、补阳还五汤高剂量组AQP4表达水平高于模型组(P<0.05)。5组脑组织含水量比较,差异有统计学意义(F=16.21,P<0.001);其中补阳还五汤高剂量组脑组织含水量低于模型组(P<0.05)。5组脑组织EB含量比较,差异有统计学意义(F=60.05,P<0.001);其中补阳还五汤低剂量组、补阳还五汤中剂量组、补阳还五汤高剂量组脑组织EB含量低于模型组(P<0.05)。结论补阳还五汤能够减轻脑出血诱发的脑水肿,其机制可能与其激活PI3K/AKT信号转导通路,调控AQP4极性表达,降低血-脑脊液屏障通透性有关。
Objective To explore the effects of Buyanghuanwu decoction on aquaporin 4（ AQP4） expression and the permeability of the blood brain barrier in cerebral hemorrhage rats. Methods From March 2014 to November 2015,SPF level adult male SD rats were randomly divided into sham operation group,model group,Buyanghuanwu decoction low,medium and high dose groups, 24 in each group. Model group and Buyanghuanwu decoction groups prepared models of cerebral hemorrhage. Since two days after modeling, Buyanghuanwu decoction low, medium, and high dose groups were given Buyanghuanwu decoction by gastric perfusion,the dose was 13. 2 g·kg^（-1）·d^（-1）,26. 5 g·kg^（-1）·d^（-1） and 53. 0 g·kg^（-1）·d^（-1） respectively,continuously for 14 days, and sham operation group and model group were given the equal volume of the corresponding 0. 9% sodium chloride solution. The expression of phosphatidylinositol 3 kinase（ PI3K） and protein kinase B（ AKT） was detected by immunohistochemistry. The expression of AQP4 was detected by immunofluorescence labeling method. Content of water in brain was detected by wet and dry weight method,and formamide method was used to detect the blood- brain barrier permeability. Results Sham operation group,model group,Buyanghuanwu decoction low,medium and high dose groups were significantly different in the expression of PI3 K and AKT（ P〈0. 05）; Buyanghuanwu decoction low,medium and high dose groups were higher than model group in the expression of PI3K; Buyanghuanwu decoction medium and high dose groups were higher than model group in the expression of AKT（ P〈0. 05）. The 5 groups were significantly different in the expression of AQP4（ F = 95. 79,P〈0. 001）; Buyanghuanwu decoction low,medium and high dose groups were higher than model group in the expression of AQP4（ P〈0. 05）. The 5 groups were significantly different in brain water content（ F= 16. 21,P〈0. 001）; Buyanghuanwu decoction high dose group was lower than model group in brain water content（ P〈0. 05）. The 5 groups were significantly different in the EB content of cerebral tissue（ F = 60. 05,P〈0. 001）; Buyanghuanwu decoction low,medium and high dose groups were lower than model group in EB content of cerebral tissue（ P〈0. 05）.Conclusion Buyanghuanwu decoction can alleviate encephaledema induced by cerebral hemorrhage,and its mechanism may be related with the activation of PI3 K / AKT singal,the regulation of the extreme expression of AQP4,and the reduction of blood-brain barrier permeability.
出处
《中国全科医学》
CAS
CSCD
北大核心
2016年第21期2557-2561,共5页
Chinese General Practice
基金
河北省高等学校重点学科病理学与病理生理学资助项目(2013)
河北省中医药抗痴呆重点研究室资助项目(2014)
承德医学院校级重点资助项目(201607)
关键词
脑出血
补阳还五汤
脑水肿
水通道蛋白质4
血脑屏障
Cerebral hemorrhage
Buyanghuanwu decoction
Brain edema
Aquaporin 4
Blood-brain barrier
