芍药苷干预施万细胞外泌体对背根神经元细胞凋亡的作用  

Effect of Paeoniflorin on Apoptosis of Dorsal Root Neurons Induced by Schwann Cell Exosomes

作　　者：韩朔 李潇 杨鑫伟[1] 朱笳悦 朱晏伯 许利平[1] Han Shuo;Li Xiao;Yang Xinwei(School of Traditional Chinese Medicine,Capital Medical University and Beijing Key Lab of TCM Collateral Disease Theory Research,Beijing 100069,China)

机构地区：[1]首都医科大学中医药学院、中医络病研究北京重点实验室,100069

出　　处：《医学研究杂志》2020年第11期49-53,共5页Journal of Medical Research

基　　金：国家自然科学基金资助项目(81873125,81704014,81473642)。

摘　　要：目的探讨芍药苷(paeoniflorin,PF)干预高糖环境施万细胞(Schwann cell,SCs)外泌体(exosomes,EXOs)对背根神经节神经元(dorsal root ganglion neurons,DRGn)细胞凋亡的影响。方法取高糖环境SCs,培养24h,透射电镜观察EXOs的形态,蛋白印迹法(Western blot)鉴定EXOs特异性标志蛋白和氧化应激的蛋白表达;将SCs来源的EXOs与DRGn共培养24h,激光共聚焦显微镜观察DRGn摄取SCs来源EXOs的情况,Western blot法检测DRGn中影响细胞凋亡蛋白的表达水平。结果EXOs形态为双层膜囊泡结构,直径在40~100nm;EXOs中标记蛋白CD63表达证实了EXOs特异性;EXOs中氧化应激蛋白表达升高,尤其是PF干预SCs来源EXOs中Nrf2、HO-1的表达升高更明显;SCs来源EXOs被DRGn摄取,分布于DRGn细胞核周围;经PF干预SCs来源EXOs影响DRGn促凋亡蛋白GADD153和Bax表达显著降低,抗凋亡蛋白Bcl-2表达明显增加。结论PF通过SCs EXOs携带氧化应激信息,影响DRGn中促凋亡蛋白表达减少,抗凋亡蛋白表达增加,从而减轻DRGn细胞凋亡,有利于改善糖尿病周围神经病变。Objective To investigate the effect of paeoniflorin on apoptosis of dorsal root neurons induced by Schwann cell exosomes in high glucose tconcentration.MethodsHigh glucose environment SCs,after 24 hours of culture,the morphology of exosomes was observed by transmission electron microscope.The expression levels of specific marker proteins and oxidative stress proteins were identified by Western blot,after the SCs source exosomes were incubated with DRGn for 24 hours,the expression level of apoptosis protein in DRGn was detected by Western blot.ResultsUnder transmission electron microscope,the exosomes were bilayer membrane vesicles with diameters between 40-100nm,and the marker proteins CD9 confirmed the specificity of EXOs.The expression of oxidative stress protein in EXOs increased,especially when PF interfered with the expression of Nrf2 and HO-1 in SCs-derived EXOs,indicating that oxidative stress was initiated.SCs-derived EXOs was absorbed by DRGn and distributed around the nucleus of DRGn.The expression of pro-apoptotic protein GADD153 and Bax was significantly decreased and the expression of anti-apoptotic protein Bcl-2 was significantly increased in DRGn treated with SCs-derived EXOs treated by PF.ConclusionPF carries oxidative stress information through SCs EXOs,which reduces the expression of pro-apoptotic proteins and increases the expression of anti-apoptotic proteins in DRGn,thus reduces the apoptosis of DRGn cells,which is beneficial to the improvement of diabetic peripheral neuropathy.

关 键 词：芍药苷 施万细胞 外泌体 背根神经节神经元 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]