毛蕊异黄酮对氧化损伤的星形胶质细胞增殖与凋亡的影响  

Effects of Pistil Isoflavones on Proliferation and Apoptosis of Oxidative Damaged Astrocytes

作　　者：吴成林 张国福[2] 许洋 郭德华[1] 刘小舟 罗露洁 WU Chenglin;ZhANG Guofu;XU Yang;GUO Dehua;LIU Xiaozhou;LUO Lujie(Jiangxi University of Traditional Chinese Medicine,Nanchang 330004,Jiangxi,China;Affiliated Hospital of Jiangxi University of Chinese Medicine,Nanchang 330006,Jiangxi,China)

机构地区：[1]江西中医药大学,江西南昌330004 [2]江西中医药大学附属医院,江西南昌330006

出　　处：《中华中医药学刊》2023年第5期151-154,I0017,I0018,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81860856);江西省级研究生创新专项(YC2021-S524);江西省校级研究生创新专项(JZYC21S23);江西中医药大学校级研究生创新专项(JZYC20S17)。

摘　　要：目的 探讨毛蕊异黄酮对氧化损伤的星形胶质细胞(astrocyte, AS)的增殖、凋亡的影响。方法 原代分离大鼠脊髓星形胶质细胞,并利用免疫荧光检测胶质纤维酸性蛋白(glial fibrillary acidic protein, GFAP)进行鉴定。分别加入5、10、20μmol·L^(-1)毛蕊异黄酮预处理脊髓星形胶质细胞12 h,然后加入过氧化氢(H_(2)O_(2))处理24 h,造成氧化损伤模型,CCK8检测各组细胞增殖情况,选择合适的毛蕊异黄酮处理浓度。实验分组包括空白对照组、模型组、模型+毛蕊异黄酮预处理组。CCK8检测各组细胞增殖情况,流式检测各组细胞凋亡和周期情况,免疫荧光检测各组细胞样本中BrdU水平,蛋白免疫印迹法(Westernblot)检测各组细胞中蛋白酪氨酸激酶2(januskinase2,JAK2)、信号转导和转录激活因子3 (signal transducers and activator of transcription3,STAT3)、糖蛋白130(recombinant glycoprotein130,GP130)、白细胞介素-6 (interleukin-6,IL-6)的蛋白表达水平。结果 H_(2)O_(2)处理能够抑制脊髓星形胶质细胞的增殖并诱导其凋亡(P<0.05),氧化损伤模型细胞组中p-JAK2、p-STAT3、GP130、IL-6的蛋白表达水平显著增加(P<0.05),而毛蕊异黄酮预处理后能够减轻H_(2)O_(2)造成的氧化损伤,促进增殖和抑制凋亡(P<0.05),并显著抑制了p-JAK2、p-STAT3、GP130、IL-6的蛋白表达水平(P<0.05)。结论 毛蕊异黄酮能够促进氧化损伤的星形胶质细胞增殖并抑制其凋亡,可能通过抑制JAK2/STAT3通路磷酸化起作用。Objective To investigate the effects of pistil isoflavones on the proliferation,cycle and apoptosis of oxidative damaged astrocytes.Methods Rat spinal cord astrocytes were isolated and identified by immunofluorescence detection of luorescencedetectionofglial fibrillary acidic protein(GFAP).Different concontrations of 5μmol/L,10μmol/L and 20μmol/L pistil isoflavones were added respectively to pretreat the medullary astrocytes for 12 h,and then H_(2)O_(2) was added for 24 h to create an oxidative damage model.CCK8 detected the cell proliferation of each group and the appropriate concentration of pistil isoflavones were selected.The experimental groups included blank control group,model group and model+pistil isoflavone pretreatment group.CCK8 was used to detect the proliferation of cells in each group.Flow cytometry was used to detect the apoptosis and cycle of cells in each group,immunofluorescence was used to detect the level of BrdU in cell samples in each group,and Western blot was used to detect the protein expression levels of phosphorylation Janu kinase 2(p-JAK2),phosphorylation signal transducer and activator of transcription 3(p-STAT3),recombinant glycoprotein 130(GP130)and interleukin-6(IL-6)in cells in each group.Results H_(2)O_(2) treatment could inhibit the proliferation and induce apoptosis of spinal cord astrocytes.The protein expression levels of p-JAK2,p-STAT3,GP130 and IL-6 in the oxidative injury model cell group were significantly increased.After pretreatment,pistil isoflavone could reduce the oxidative damage caused by H_(2)O_(2),promote the proliferation and inhibit the apoptosis,and significantly inhibit the protein expressions of p-JAK2,p-STAT3,GP130 and IL-6.Conclusion Pistil isoflavone can promote the proliferation and inhibit the apoptosis of oxidative damaged astrocytes,which may play a role by inhibiting the phosphorylation of JAK2/STAT3 pathway.

关 键 词：星形胶质细胞 毛蕊异黄酮 Janu激酶2/信号转导和转录活化因3 增殖 凋亡 

分 类 号：R285.5[医药卫生—中药学]