紫云英苷调控炎性痛模型小鼠L_(4-5)脊髓背角星形胶质细胞自噬与凋亡  

Astragalin Regulates Autophagy and Apoptosis of Astrocytes in L_(4-5)Spinal Dorsal Horn of Mouse Inflammatory Pain Model

作　　者：张维珊 林佳泓 王灿 张润恒 杨俊华 刘靖[1] 李国营[1,2] 马宇昕[1] ZHANG Weishan;LIN Jiahong;WANG Can;ZHANG Runheng;YANG Junhua;LIU Jing;LI Guoying;MA Yuxin(Department of Human Anatomy,Histology and Embryology,School of Basic Medical Sciences,Guangdong Pharmaceutical University,Guangzhou 510006,China;Guangdong Medical Association,Guangzhou 510180,China)

机构地区：[1]广东药科大学基础医学院人体解剖学与组织胚胎学系,广东广州510006 [2]广东省医学会,广东广州510180

出　　处：《中山大学学报(医学科学版)》2025年第2期186-196,共11页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省医学科学技术研究基金(B2022076);广东省中医药局中医药科研项目(20231201);广东省普通高校特色创新类项目(2023KTSCX057)。

摘　　要：【目的】探究紫云英苷(AST)对完全弗氏佐剂(CFA)诱发的炎性痛小鼠L_(4-5)脊髓背角星形胶质细胞自噬及凋亡的影响。【方法】将24只雄性6月龄C57BL/6小鼠随机分为正常组(control)、生理盐水组(saline)、CFA模型组、CFA+AST组,每组6只小鼠。于小鼠右侧腓骨外踝窝注射10μL CFA建立炎性痛小鼠模型。saline组小鼠注射等量生理盐水。CFA+AST组小鼠在疼痛造模成功后腹腔注射给药60 mg/kg AST,每日一次,连续给药21 d。采用多重免疫荧光染色法检测各组小鼠L_(4-5)脊髓背角自噬相关因子ATG12、Beclin-1及凋亡相关因子Cleaved-Caspase 3、Caspase 9与星形胶质细胞标志物GFAP的共表达情况;Western blot法检测各组小鼠L_(4-5)脊髓背角自噬相关蛋白ATG12、Beclin-1及凋亡相关蛋白Caspase 3、Caspase 9表达水平。【结果】免疫荧光染色结果表明,与CFA模型组相比,CFA+AST组小鼠L_(4-5)脊髓背角ATG12(P<0.0001)、Beclin-1(P<0.0001)荧光强度增强,Cleaved-Caspase 3(P<0.001)、Caspase 9(P<0.0001)荧光强度下降,并且AST可抑制星形胶质细胞活化。Western blot结果进一步证实,AST显著上调CFA小鼠L_(4-5)脊髓背角ATG12(P<0.0001)、Beclin-1(P<0.0001)表达,并下调Caspase 3(P<0.01)、Caspase 9(P<0.001)表达。【结论】AST促进CFA小鼠L_(4-5)脊髓背角星形胶质细胞自噬并抑制其凋亡。【Objective】To explore the effects of astragalin(AST)on autophagy and apoptosis of astrocytes in the L_(4-5)dorsal horn of the spinal cord in mice with inflammatory pain induced by complete Freund's adjuvant(CFA).【Methods】Twenty-four male C57BL/6 mice,aged six months,were randomly assigned to four groups:control group,saline group,CFA model group,and CFA+AST group,six mice in each group.The inflammatory pain model was established by injection of 10μL CFA into the right lateral malleolus fossa.The saline group were injected with an equal amount of normal saline at the same site.The inflammatory pain mice in CFA+AST group were further treated with AST(60 mg/kg)intraperitoneally once a day for 21 consecutive days.Multiplex immunofluorescence staining was used to detect the coexpression of autophagy-related factors including ATG 12 and Beclin-1,apoptosis-related factors including Cleaved Caspase 3 and Caspase 9,and the astrocyte marker such as GFAP in the L_(4-5)spinal dorsal horn of the mice in each group.Western blot was used to examine the protein expression levels of autophagy-related proteins(ATG 12,Beclin-1)and apoptosis-related proteins(Caspase 3,Caspase 9)in the L_(4-5)spinal dorsal horn of mice.【Results】Immunofluorescent staining showed that in the L_(4-5)dorsal horn of the spinal cord,the fluorescence intensity of ATG 12(P<0.0001)and Beclin-1(P<0.0001)was significantly increased,while that of Cleaved-Caspase 3(P<0.001)and Caspase 9(P<0.0001)was decreased in the CFA+AST group when compared to the CFA model group.Furthermore,AST could inhibit the activation of astrocytes.Western blot further confirmed that AST significantly upregulated the expression of ATG 12(P<0.0001)and Beclin-1(P<0.0001)in the L_(4-5)spinal cord of CFA mice,and downregulated the expression of Caspase 3(P<0.01)and Caspase 9(P<0.001).【Conclusions】AST promotes autophagy of astrocytes and inhibits their apoptosis in the L_(4-5)spinal dorsal horn of CFA mice.

关 键 词：紫云英苷 炎症性疼痛 脊髓背角 星形胶质细胞 自噬 凋亡 

分 类 号：R285.5[医药卫生—中药学]