MANF对A2型反应性星形胶质细胞增殖、凋亡的影响  

Effects of MANF factor on the proliferation and apoptosis of A2 type reactive astrocytes

作　　者：刘锐 丁振飞 戴策 钟林[1] 汪萧和 尹宗生[1] 高维陆[1] Liu Rui;Ding Zhenfei;Dai Ce(Dept of Orthopedics,The First Affiliated Hospital of Anhui Medical University,Hefei 230022)

机构地区：[1]安徽医科大学第一附属医院关节外科,合肥230022

出　　处：《安徽医科大学学报》2021年第10期1511-1515,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81871785、81672161)。

摘　　要：目的探讨中脑星形胶质细胞源性神经营养因子(MANF)对脊髓A2型反应性星形胶质细胞(RAS)增殖及凋亡的影响。方法取SD大鼠脊髓组织进行星形胶质细胞的分离及培养,并采用免疫荧光技术鉴定细胞。采用白细胞介素-1β(IL-1β)刺激星形胶质细胞制备A2型RAS,作为实验组,并用未处理组做空白对照组。然后采用免疫荧光染色法对制备的A2型RAS进行鉴定。将制备完成的细胞分成Control组和MANF组,以不同浓度(0、0.5、1、2μg/ml)的MANF蛋白及不同的作用时间(12、24、36、48 h)处理细胞,采用CCK-8法检测在450 nm处的吸光度。取一定浓度的MANF蛋白(1μg/ml)处理细胞,同样分为Control组和MANF组,采用EdU法验证对A2型RAS增殖能力的影响。采用TUNEL法验证对A2型RAS凋亡的影响。结果免疫荧光结果显示IL-1β可以刺激星形胶质细胞向A2型转化;CCK-8、EdU增殖实验结果显示,MANF蛋白干预后,细胞增殖能力均显著高于对照组(P<0.05);TUNEL凋亡实验显示经MANF蛋白干预后,凋亡细胞明显减少(P<0.05)。结论MANF蛋白对A2型RAS具有促进增殖、抑制凋亡的作用。Objective To study the effect of mesencephalic astrocyte-derived neurotrophic factor(MANF)on the proliferation and apoptosis of A2 type reactive astrocytes(RAS)in the spinal cord.Methods The spinal cord of SD rats was taken to separate and culture astrocytes,and the cells were identified by immunofluorescence technology.Interleukin-1β(IL-1β)was used to stimulate astrocytes to prepare A2 type RAS as the experimental group;and the untreated group was used as a blank control group.Immunofluorescence was used to identify A2 type RAS.The prepared cells were divided into control group and MANF group.The cells were treated with different concentrations(0,0.5,1,2μg/ml)of MANF protein and different action times(12,24,36,48 h),and CCK-8 method was used to detectthe absorbance at 450 nm.The cells were treated with a certain concentration of MANF protein(1μg/ml)and divided into the control group and the MANF group.The EdU method was used to verify the effect on the proliferation of A2 RAS.The TUNEL method was used to verify the effect on A2 RAS apoptosis.Results Immunofluorescence results showed that IL-1βcould stimulate the transformation of astrocytes to type A2;The results of CCK-8 and EdU proliferation experiments showed that after MANF protein intervention,the cell proliferation ability was significantly higher than that of the control group(P<0.05);TUNEL apoptosis experiment showed that after the intervention of MANF protein,apoptotic cells significantly reduced(P<0.05).Conclusion MANF protein can promote proliferation and inhibit apoptosis of A2 RAS.

关 键 词：中脑星形胶质细胞源性神经营养因子 A2型反应性星形胶质细胞 增殖 凋亡 

分 类 号：R651.2[医药卫生—外科学]