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作 者:温智林 王真真 贺文彬[1,2] 陈乃宏[1,2,3]
机构地区:[1]山西中医学院,中医脑病学山西省重点实验室&脑藏象学山西省高等学校重点实验室 [2]中国医学科学院,药物研究所&神经科学中心 [3]湖南中医药大学
出 处:《神经药理学报》2014年第6期1-5,共5页Acta Neuropharmacologica
基 金:国家自然科学基金项目(No.81274122;No.81202507;No.81273629;No.81473375);教育部博士点基金重点项目(No.20121106130001);新药作用机制研究与药效评价北京市重点实验室资助项目(No.BZ0150);中央级公益性科研院所基本科研业务费;专项资金项目(No.2014RC03);山西省回国留学人员科研资助项目(No.2013-134);山西省国际科技合作项目(No.2010081065)
摘 要:目的:观察开心散含药血清对皮质酮(corticosterone)损伤大鼠大脑皮质星形胶质细胞CTX TNA2的影响。方法:制备开心散含药血清(500 mg·kg-1)、阳性药氟西汀含药血清(10 mg·kg-1)、普通大鼠血清。细胞实验中采用皮质酮损伤CTX TNA2细胞建立体外抑郁模型,MTT法检测开心散对皮质酮所致细胞损伤的影响以及磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)信号通路抑制剂的干预作用;采用Western Blotting法检验相关信号通路蛋白表达变化情况。结果:皮质酮浓度为200μmol·L-1时,细胞活力抑制率可稳定在60%,浓度依赖性较好;开心散含药血清均能够显著提高皮质酮损伤细胞的存活率,而给予PI3K相关通路的抑制剂后,开心散保护作用被逆转,Western Blotting结果显示给予开心散含药血清能够逆转皮质酮损伤后PI3K及蛋白激酶B(protein kinase B,AKT)1/2蛋白表达下降的趋势。结论:开心散含药血清对皮质酮所致的CTX TNA2细胞损伤具有明显保护作用,其保护机制可能与PI3K信号通路有关。Objective: To observe the effects of Kaixin powder containing serum on corticosterone induced injury of rat cortical astrocyte CTX TNA2 cell line. Methods: Firstly, SD rats were orally administered with Kaixin powder, positive drug fluoxetine (10 mg.kg-1) and solvent control in order to obtain corresponding serum. CTX TNA2 cell line injured by corticosterone was used to establish depression model in vitro and cell activity was detected with MTT. Then we tested whether Kaixin powder play a neuroprotective role through PI3K signaling pathway in this cell model with the inhibitors of this pathway and detected by western blotting analysis. Results: According to our data, when the corticosterone concentration reached 200 μmol.L-1, the cell viability inhibition rate was stabilized at 60% with a well concentration dependent. Kaixin powder containing serum could significantly improve the survival rate of corticosterone injured CTX TNA2 cells. The western blotting results also showed that Kaixin powder containing serum could reverse the reduction of PI3K and AKT1/2 protein expression induced by corticosterone. However, after given the inhibitors of PI3K pathway, the protective effect of Kaixin powder was reversed. Conclusion: Kaixin powder containing serum has obvious protective effect on corticosterone injured CTX TNA2 cell line, and the mechanism may be associated with nerve cell protective related PI3K signal transduction pathway.
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