健脾补土组方对体外神经元缺氧损伤后的凋亡情况及INT、ILK、FAK表达的影响  

Effects of Invigorating Spleen and Tonifying Earth Formula on Rat Neuron Apoptosis Situation and Expressions of INT,ILK and FAK after Rat Neuron Lack of Blood and Oxygen

作　　者：李花[1,2,3] 陈昱文[1,2] 刘旺华[1,2,3] 周小青[1,2,3] 郑彩杏 金梦[1,2] 范元硕 李诚均[1,2] 颜艳艳 

机构地区：[1]湖南中医药大学中医诊断研究所,湖南长沙410208 [2]湖南中医药大学中医诊断学湖南省重点实验室,湖南长沙410007 [3]湖南中医药大学数字中医药协同创新中心,湖南长沙410208

出　　处：《中华中医药学刊》2018年第3期538-542,I0004-I0006,共8页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81202632,81373702,81473567);教育部博士点基金项目(20124323120003);湖南省研究生科研创新项目(CX2016B368);湖南省自然科学基金项目(13JJ3097);湖南省教育厅科研项目(14B134);湖南省高校创新平台开放基金项目(15K092);湖南中医药大学中诊开放基金项目(2015ZYZD30)

摘　　要：目的：观察健脾补土组方对新生SD鼠神经元体外培养缺氧造模后的凋亡情况以及INT、ILK、FAK表达的影响,探讨其减少神经元凋亡的作用机制。方法：原代分离培养新生SD鼠神经元,接种于六孔板中,并分为正常血清对照组、正常血清＋低氧组、低氧＋正常血清＋神经生长因子组、低氧＋健脾补土组方含药血清组、低氧＋健脾补土组方含药血清＋整合素抗体组。将标本放入37℃三气培养箱中,诱导24 h后,恢复正常条件培养4h。用流式细胞术检测检测神经元凋亡,RT-qPCR法检测INT、ILK、FAK mRNA的表达,用免疫细胞化学法和Western blot法检测INT、ILK、FAK蛋白的表达。结果：与正常对照组比较,其余各组神经元凋亡率均显著增加（P〈0.01）,INT、ILK、FAK mRNA及蛋白表达显著减少（P〈0.01）;与低氧模型组比较,健脾补土组INT、ILK、FAK mRNA及蛋白表达显著增强（P〈0.01）。结论：健脾补土组方能够减少缺氧损伤后神经元细胞凋亡,其作用机制可能与其通过上调INT、ILK、FAK表达有关。Objective： To study the effects of Invigorating Spleen and Tonifying Earth Formula on rat neuron apoptosis situation and expressions of INT,ILK and FAK after rat neuron lack of blood and oxygen. To explore the mechanism of reducing the neuron apoptosis. Methods： We separated and cultivated the neuron of newborn SD rats which were assigned to six orifice plates. These plates were divided into control group,hypoxia group,neural factor group,Invigorating Spleen and Tonifying Earth Formula group and integrin antibody group. Take these specimens into three gas incubators at 37 ℃. After inducing for 24 h,and cultivating for 4 h at the normal conditions. We used flow cytometry to detect neuron apoptosis and RT-q PCR and Western blot respectively to detect the mRNA and protein expressions of INT,ILK and FAK. Results：Compared with control group,the rest of the group neurons apoptosis rate were significantly increased（ P 0. 01）,and expressions of INT,ILK,FAK mRNA and protein reduced significantly（ P 0. 01）. Compared with hypoxia group,the expressions of INT,ILK,FAK mRNA and protein significantly increased in Invigorating Spleen and tonifying earth group（ P 0. 01）. Compared with Invigorating Spleen and Tonifying Earth Formula group,the expressions of INT,ILK,FAK mRNA and protein significantly reduced in neural factor group and integrin antibody group（ P 0. 01）. Conclusion： In-vigorating Spleen and Tonifying Earth Formula can reduce the neuron apoptosis after hypoxia. The mechanism may be related to increasing the expressions of INT,ILK and FAK.

关 键 词：健脾补土 神经元 凋亡 整合素 整合素连接激酶 黏着斑激酶 

分 类 号：R285.5[医药卫生—中药学]