芍药苷对糖尿病周围神经病变状态下线粒体输入途径蛋白TOM20的作用  

Effect of paeoniflorin on TOM20 of mitochondrial import pathway in diabetic peripheral neuropathy

作　　者：刘东齐 闫仕祺 刘浩龙 杨鑫伟[1] LIU Dongqi;YAN Shiqi;LIU Haolong;YANG Xinwei(Beijing Key Laboratory of Traditional Chinese Medicine Research on Collaterals,School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China;Beijing Chao-Yang Hospital,Capital Medical University,Beijing 100020,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 [2]首都医科大学附属北京朝阳医院,北京100020

出　　处：《中草药》2024年第9期2987-2995,共9页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金面上项目(82174184);北京市教育委员会科技计划一般项目(KM202210025021);北京市科学技术协会青年人才托举工程(BYESS2023386);国家自然科学基金青年项目(82204826)。

摘　　要：目的探究芍药苷对糖尿病周围神经病变(diabetic peripheral neuropathy,DPN)大鼠坐骨神经和高糖环境下雪旺细胞(Schwann cells,SCs)线粒体输入途径蛋白线粒体外膜转位酶20(translocase of outer mitochondrial membrane 20,TOM20)的影响。方法以25、100、150 mmol/L葡萄糖分别干预RSC96雪旺细胞株12、24、48 h,用高内涵分析法检测TOM20和线粒体硫氧还蛋白2(mitochondrial thioredoxin 2,Trx2)的表达;设置对照组(给予25 mmol/L葡萄糖)、TOM20 siRNA组、TOM20 siRNA+芍药苷24 h组和TOM20 siRNA+芍药苷48 h组,转染TOM20 siRNA后给予10 mmol/L芍药苷分别干预24、48 h,利用Western blotting检测TOM20和Trx2蛋白表达;设置对照组(给予25 mmol/L葡萄糖)、高糖组(给予150 mmol/L葡萄糖)和芍药苷组(给予150 mmol/L葡萄糖和10 mmol/L芍药苷),利用高内涵分析法检测TOM20和Trx2蛋白表达。制备DPN大鼠模型,给予白芍总苷后,利用免疫荧光法检测大鼠坐骨神经中TOM20和Trx2蛋白表达。结果高内涵分析结果显示,150 mmol/L葡萄糖干预12、24 h后,TOM20和Trx2蛋白表达水平均显著降低(P<0.05、0.01),且2种蛋白的表达变化存在相关性。与对照组比较,高糖组TOM20和Trx2蛋白表达均显著降低(P<0.01);与高糖组比较,芍药苷组TOM20和Trx2蛋白表达均显著升高(P<0.01)。Western blotting结果显示,与对照组比较,TOM20 siRNA组TOM20和Trx2蛋白表达水平均显著降低(P<0.05、0.01);与TOM20 siRNA组比较,芍药苷干预24、48 h后TOM20和Trx2蛋白表达水平均显著升高(P<0.05、0.01)。与对照组比较,模型组大鼠坐骨神经中TOM20和Trx2蛋白表达均显著降低(P<0.01);与模型组比较,白芍总苷组TOM20和Trx2蛋白表达均显著升高(P<0.01)。结论芍药苷能够通过上调SCs以及大鼠坐骨神经中TOM20蛋白的表达,促进Trx2蛋白的线粒体输入,从而有效对抗线粒体氧化应激干预DPN。Objective To investigate the effect of paeoniflorin(PF)on the protein involved in mitochondrial import pathway mitochondrial translocase of outer mitochondrial membrane 20(TOM20)in the sciatic nerve of diabetic peripheral neuropathy(DPN)rats and high glucose environment of Schwann cells(SCs).Methods RSC96 cells was used to intervene with 25,100 and 150 mmol/L glucose for 12,24 and 48 h,respectively,and the expressions of TOM20 and mitochondrial thioredoxin 2(Trx2)were detected by high content analysis;Control group(25 mmol/L glucose),TOM20 siRNA group,TOM20 siRNA+paeoniflorin 24 h group,and TOM20 siRNA+paeoniflorin 48 h group were set up.After transfection with TOM20 siRNA,10 mmol/L paeoniflorin was given for 24,48 h,respectively,Western blotting was used to detect TOM20 and Trx2 protein expressions;Control group(25 mmol/L glucose),high glucose group(150 mmol/L glucose),and paeoniflorin group(150 mmol/L glucose+10 mmol/L paeoniflorin)were set up,high connotation analysis was used to detect TOM20 and Trx2 protein expressions.DPN rat model was prepared and given total glucosides of paeony,immunofluorescence was used to detect TOM20 and Trx2 protein expressions in sciatic nerve of rats.Results The results of high content analysis showed that after 12,24 h of 150 mmol/L glucose intervention,TOM20 and Trx2 protein expressions were significantly decreased(P<0.05,0.01),and there was a correlation between the expressions of the two proteins.Compared with control group,TOM20 and Trx2 protein expressions in high glucose group were significantly decreased(P<0.01);Compared with high glucose group,TOM20 and Trx2 protein expressions in paeoniflorin group were significantly increased(P<0.01).The results of Western blotting showed that compared with control group,TOM20 and Trx2 protein expressions in TOM20 siRNA group were significantly decreased(P<0.05,0.01);Compared with TOM20 siRNA group,TOM20 and Trx2 protein expressions were significantly increased after intervention with paeoniflorin for 24,48 h(P<0.05,0.01).Compared with

关 键 词：芍药苷 雪旺细胞 糖尿病周围神经病变 线粒体膜外转位酶20 线粒体硫氧还蛋白 

分 类 号：R285.5[医药卫生—中药学]