体外培育牛黄配伍冰片激活PI3K/Akt通路抑制脑缺血再灌注模型大鼠神经元凋亡  

In Vitro Cultured Calculus Bovis Combined with Borneol Activates PI3K/Akt Pathway and Inhibit Neuronal Apoptosis in Cerebral Ischemia-Reperfusion Model Rats

作　　者：陈海 任敉宏 郭晓庆 李勇 李红燕 马荣 谢倩[1] 王建[1] Chen Hai;Ren Mihong;Guo Xiaoqing(National Key Laboratory of Southwestern Chinese Medicine Resources,Key Laboratory of Standardization of Traditional Chinese Medicine Materials of Ministry of Education,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Department of Pharmacy,Changzhi Medical College,Changzhi 046000,China;Beijing Ruinuo Zhongde Technology Limited Company,Beijing 100089,China)

机构地区：[1]成都中医药大学药学院,西南特色中药资源国家重点实验室,中药材标准化教育部重点实验室,成都611137 [2]长治医学院药学院,长治046000 [3]北京瑞诺众德科技有限公司,北京100089

出　　处：《华中科技大学学报（医学版）》2023年第6期816-822,共7页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金面上项目(No.81873023);山西省高等学校科技创新项目(No.2022L371);长治医学院博士科研启动基金项目(No.BS202208)。

摘　　要：目的从抗凋亡角度探讨体外培育牛黄(ICCB)、冰片及其配伍对脑缺血再灌注模型大鼠脑保护作用的分子机制。方法采用改良线栓法制备短暂大脑中动脉阻塞(MCAO)模型,缺血90 min后再灌注,苏木精-伊红(HE)染色观察缺血侧脑组织病理损伤情况;Tunel染色观察缺血侧大脑皮层神经元凋亡;RT-PCR检测相关蛋白mRNA的表达;Western blot及免疫组织化学方法检测PI3K/Akt通路相关蛋白的表达。结果与溶剂模型组比较,体外培育牛黄、冰片及两药配伍可不同程度地降低MCAO模型大鼠缺血侧脑组织病理损伤,抑制缺血侧皮层神经元凋亡,升高缺血侧脑组织中p-PI3K、p-Akt及Bcl-2蛋白表达,降低Caspase-3、Bax蛋白表达以及Bax/Bcl-2比值。结论体外培育牛黄、冰片及二者配伍的脑神经保护作用与激活PI3K/Akt通路,抑制脑缺血再灌注后神经元凋亡有关,进而发挥“开窍醒神”功效。Objective Based on PI3K/Akt signaling pathway,the molecular mechanism of brain protective effect of in vitro cultured Calculus Bovis(ICCB),borneol and their combination on cerebral ischemia-reperfusion model rats was discussed from the perspective of anti-apoptosis.Methods The model of transient middle cerebral artery occlusion(MCAO)was established by modified suture and reperfusion 90 minutes after ischemia.HE staining and Tunel staining were used to observe the pathological injury of brain tissue and the apoptosis of cerebral cortical neurons.The expression levels of PI3K/Akt pathway related proteins were detected by Western blotting and immunohistochemistry.Expression of mRNA was detected by RT-PCR.Results Compared with the solvent model group,the pathological injury of brain tissue was reduced and apoptosis of cortical neurons on the ischemic side was inhibited;the expressions of p-PI3K,p-Akt and Bcl-2 protein in the ischemic brain tissue were increased;the expression of Caspase-3,Bax protein and Bax/Bcl-2 were decreased in ICCB group,borneol group and combination group.Conclusion The brain protective effect of ICCB,borneol and their combination is related to activation of PI3K/Akt pathway and inhibition of neuronal apoptosis after cerebral ischemia-reperfusion,so as to play the role of"opening the orifices and waking the mind".

关 键 词：体外培育牛黄 冰片 脑缺血再灌注 PI3K/AKT信号通路 凋亡 

分 类 号：R743[医药卫生—神经病学与精神病学]