五苓散同方汤剂缺失成分对氧糖剥夺后星形胶质细胞水肿及水通道蛋白4表达的影响  

Effects of 5 main missing chemical components in the decoction of Wuling Powder on swelling of astrocyte and expression of AQP4 after oxygen-glucose deprivation

作　　者：吴桐 孙彩霞[1] 王建农[1] 刘良裕 杨宇珂 WU Tong;SUN Cai-xia;WANG Jian-nong;LIU Liang-yu;YANG Yu-ke(Beijing Key Laboratory of Chinese Materia Pharmacology,Institute of Basic Medical Sciences of Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China)

机构地区：[1]中国中医科学院西苑医院基础医学研究所/中药药理北京市重点实验室,北京100091

出　　处：《中国新药杂志》2021年第6期540-545,共6页Chinese Journal of New Drugs

基　　金：国家自然科学基金资助项目(30873359)。

摘　　要：目的:探讨5种五苓散同方汤剂缺失成分对氧糖剥夺并复氧后星形胶质细胞水肿及其水通道蛋白4(AQP4)表达的影响。方法:星形胶质细胞分为正常组、模型组、桂皮醛(CIN)组、23-乙酰泽泻醇B(ALB)组、麦角甾醇(ERG)组、茯苓酸(PA)组、白术内酯Ⅰ(ATR)组、成分组合(MCC)组及奥扎格雷(OZA)组。用MTT法评价对细胞的抑制率,以乳酸脱氢酶(LDH)漏出率反映细胞的损伤程度,用Western Blot法测定AQP4的表达变化。结果:氧糖剥夺2 h后复氧6 h,LDH漏出率明显增加(P<0.01),浓度10μg·mL^(-1)的ALB,0.01μg·mL^(-1)的PA和ATR及0.1μg·mL^(-1)的MCC的干预能明显降低氧糖剥夺后复氧LDH漏出率增高的程度(P<0.01)。氧糖剥夺后复氧条件下,各组AQP4表达存在明显差异。与正常组相比,模型组AQP4在氧糖剥夺后复氧条件下蛋白表达显著增加(P<0.01);与模型组相比,CIN,ALB,ERG,PA,ATR及MCC组的AQP4在氧糖剥夺后复氧条件下的蛋白表达变化趋势相似,但CIN,ALB,ERG,PA,ATR及MCC组AQP4蛋白表达均明显降低(P<0.01)。结论:五苓散同方汤剂5种主要缺失成分能显著减轻2 h氧剥夺后复氧6 h的星形胶质细胞水肿,机制可能与其通过降低AQP4蛋白表达水平有关。Objective:To investigate the effects of five missing components in the Decoction of Wuling Powder on astrocyte edema and aquaporin4(AQP4)expression after oxygen glucose deprivation and reoxygenation.Methods:Astrocytes were divided into normal group,model group,cinnamaldehyde(CIN)group,alisol B 23-acetate(ALB)group,ergosterol(ERG)group,pachymic acid(PA)group,atractylenolideⅠ(ATR)group,medicinal chemical composition(MCC)group and ozagrel(ozagrel,OZA)group.MTT method was used to evaluate the drug’s inhibitory rate on the cells,the lactate dehydrogenase(LDH)leakage rate was used to reflect the degree of cell damage,and Western blot method was used to determine the expression change of AQP4.Results:After 2 hours of oxygen glucose deprivation and 6 hours of reoxygenation,the LDH leakage rate was significantly increased(P<0.01).ALB at a concentration of 10μg·mL^(-1),PA and ATR at 0.01μg·mL^(-1),and MCC at 0.1μg·mL^(-1)could significantly reduce the increase in the leakage rate of reoxygenation LDH after oxygen glucose deprivation(P<0.01).The expression of AQP4 under oxygen and glucose deprivation and reoxygenation conditions showed significant differences among the groups.Compared with the normal group,the expression of AQP4 in the model group was significantly increased after oxygen glucose deprivation under reoxygenation conditions(P<0.01).Compared with the model group,the expression trends of AQP4 in the CIN,ALB,ERG,PA,ATR and MCC groups under reoxygenation conditions after oxygen glucose deprivation were similar,but the AQP4 protein expressions in the CIN,ALB,ERG,PA,ATR,and MCC groups were all significantly reduced(P<0.01).Conclusion:The five main missing components in the decoction of Wuling Powder can significantly reduce the astrocyte edema after 2 hours of oxygen deprivation and 6 hours of reoxygenation,which may be related to the decrease of AQP4 protein expression level.

关 键 词：五苓散 星形胶质细胞 成分缺失 AQP4 脑水肿 

分 类 号：R932[医药卫生—生药学]