芍药苷调节高糖环境下线粒体相关内质网膜对雪旺细胞凋亡的影响  

Effect of paeoniflorin on schwann cells apoptosis by regulating mitochondria-associated endoplasmic reticulum membranes in high glucose environment

作　　者：孙晓萌 李潇 朱笳悦 韩朔 杨鑫伟[1] 许利平[1] SUN Xiaomeng;LI Xiao;ZHU Jiayue;HAN Shuo;YANG Xinwei;XU Liping(School of Traditional Chinese Medicine,Capital Medical University,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,中医络病研究北京市重点实验室,北京100069

出　　处：《环球中医药》2020年第6期964-969,共6页Global Traditional Chinese Medicine

基　　金：国家自然基金青年科学基金(81704014),国家自然科学基金面上项目(81473142,81873125)。

摘　　要：目的探讨芍药苷(paeoniflorin,PF)调节高糖环境下线粒体相关内质网膜(mitochondria-associated endoplasmic reticulum membranes,MAMs)干预雪旺细胞(schwann Cell,SCs)凋亡的机制。方法采用高糖环境下的SCs模型,观察PF对高糖环境下24和48小时RSC96细胞的影响;设立空白对照组,模型对照组,PF低、中、高剂量组;采用激光共聚焦法观察MAMs的形态并检测其含量;采用流式细胞术检测细胞内的钙离子浓度(Ca 2+);采用Western blot法定量检测MAMs中线粒体融合蛋白2(Mitofusin2,Mfn2)的表达;采用Elisa法检测MAMs中蛋白激酶R样内质网激酶(PKR-like endoplasmic reticulum kinase,PERK)的表达。结果干预24和48小时后,(1)模型对照组SCs形态由棱形变为不规则圆形,其MAMs的含量和质量均比空白对照组下降(P<0.01),不同浓度的PF组SCs形态有所改善,MAMs的含量与质量均比模型对照组有明显提升(P<0.01);(2)模型对照组Mfn2表达比空白对照组明显下降(P<0.01),PF低、中、高剂量组Mfn2表达均比模型对照组明显提升(P<0.01);(3)模型对照组PERK表达比空白对照组明显升高(P<0.01),PF低、中、高剂量组PERK表达比模型对照组显著下降(P<0.05或P<0.01);(4)模型对照组细胞内Ca 2+比空白对照组明显提高(P<0.01),PF低、中、高剂量组比模型对照组均显著降低(P<0.01)。结论PF能够通过上调Mfn2的表达并抑制PERK表达,增加SCs内MAMs数量,改善SCs形态,降低细胞内Ca 2+,减轻钙超载,从而减少SCs凋亡,有利于改善糖尿病周围神经病变。Objective To explore the effect of paeoniflorin(PF)on schwann cells(SCs)apoptosis by regulating mitochondria-associated endoplasmic reticulum membranes(MAMs)in high glucose environment.Methods The schwann cells model in high glucose environment was used to observe the effects of paeoniflorin on RSC96 cells in 24 and 48 hours in high glucose environment.The blank control group,the model control group and the low,medium and high doses of paeoniflorin(PF1,PF10,PF100)group were given.The morphology of MAMs was observed by laser confocal method and its content was detected.The intracellular Ca 2+concentration was detected by flow cytometry.The expression of mitofusion 2(Mfn2)in MAMs was detected by western blot.The expression of protein kinase R-like endoplasmic reticulum kinase(PERK)in MAMs was detected by ELISA.Results After 24h and 48h intervention,the results are as follow:(1)The morphology of schwann cells in the model group changed from prismatic to irregularly round,and the content and quality of MAMs decreased compared with the blank group(P<0.01).The morphology of schwann cells in different concentrations of paeoniflorin group was improved,and the quantity and quality of MAMs were increased in different degrees compared with the model group(P<0.01).(2)The expression of Mfn2 in model group was significantly lower than that in blank group(P<0.01).The expression of Mfn2 in different concentrations of paeoniflorin groups were significantly higher than that in model group(P<0.01).(3)The expression of PERK in model group was significantly higher than that in blank group(P<0.01).Compared with model group,the expression of PERK protein in different concentrations of paeoniflorin groups decreased significantly,the difference were statistically significant(P<0.05 or P<0.01).(4)The concentration of intracellular Ca 2+in model group was significantly higher than that in blank group(P<0.01).The concentrations of paeoniflorin in different concentration groups were lower than that in model group(P<0.01).Conclusion Paeoniflo

关 键 词：芍药苷 雪旺细胞 线粒体相关内质网膜 线粒体融合蛋白2 蛋白激酶R样内质网激酶 

分 类 号：R285.5[医药卫生—中药学]