β-细辛醚对氧糖剥夺/复糖复氧诱导SH-SY5Y细胞损伤的作用机制研究  

Study on the mechanism of β-asarone on OGD/R-induced SH-SY5Y cell damage

作　　者：郭美彤 徐飞飞 李晓青 韩诚 张益铭 田雅娟 李钦青 楚世峰[2] 贺文彬 GUO Mei-tong;XU Fei-fei;LI Xiao-qing;HAN Cheng;ZHANG Yi-ming;TIAN Ya-juan;LI Qin-qing;CHU Shi-feng;HE Wen-bin(Shanxi University of Chinese Medicine,Shanxi Key Laboratory of Chinese Medicine Encephalopathy,Taiyuan 030024,China;Institute of Materia Medica,Chinese Academy of Medical Sciences,Beijing 100050,China)

机构地区：[1]山西中医药大学,中医脑病学山西省重点实验室,太原030024 [2]中国医学科学院药物研究所,北京100050

出　　处：《中华中医药杂志》2021年第9期5573-5576,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81473375);山西省重点研发计划(国际科技合作)重大区域创新合作项目(No.201803D421006,No.201903D421018);山西省回国留学人员科研项目(No.2013-134);山西省留学回国人员科技活动择优资助项目。

摘　　要：目的:研究β-细辛醚对氧糖剥夺/复糖复氧(OGD/R)诱导人源性神经母细胞瘤细胞(SH-SY5Y)损伤的影响。方法:SH-SY5Y细胞随机分为正常对照组、OGD/R组、β-细辛醚(10、20、40、80μg/mL)组和依达拉奉(15μg/mL)组。CCK-8法检测细胞存活率,微量酶标法检测乳酸脱氢酶(LDH)释放率,以确定β-细辛醚最佳作用剂量;倒置相差显微镜观察细胞形态;ELISA法检测肿瘤坏死因子-α(TNF-α);免疫印迹法检测肿瘤坏死因子受体-1(TNF-R1)蛋白表达水平。结果:与正常对照组比较,OGD/R组的细胞存活率显著下降(P<0.01),LDH释放率显著增加(P<0.01),细胞形态明显受损,TNF-α浓度和TNF-R1蛋白表达水平显著增加(P<0.01)。与OGD/R组比较,β-细辛醚(20、40、80μg/mL)均可增加细胞存活率和降低LDH释放率(P<0.01),其中β-细辛醚20μg/mL作用最佳。β-细辛醚20μg/mL可使细胞形态得到改善,细胞内TNF-α浓度和TNF-R1蛋白表达水平均显著降低(P<0.05)。结论:β-细辛醚能够改善OGD/R诱导SH-SY5Y细胞损伤,并且该作用机制可能是通过抑制OGD/R损伤中的TNF-α和TNF-R1实现改善神经损伤。Objective: To study the effects of β-asarone on human neuroblastoma cells(SH-SY5 Y) injury induced by oxygen glucose deprivation/reoxygenation(OGD/R). Methods: SH-SY5 Y cells were randomly divided into normal control group,OGD/R model group, β-asarone(10, 20, 40, 80μg/mL) group and Edaravone(15μg/mL) group. CCK-8 method was used to detect cell survival rate, and microenzyme method was used to detect the release rate of lactate dehydrogenase(LDH) to determine the optimal dose of β-asarone. Cell morphology was observed using an inverted phase contrast microscope;tumor necrosis factor-α(TNF-α) was detected by ELISA;and the expression of tumor necrosis factor receptor-1(TNF-R1) protein was detected by immunoblotting. Results: Compared with the normal control group, the cell survival rate of the OGD/R group was significantly decreased(P<0.01), the LDH release rate was significantly increased(P<0.01), the cell morphology was significantly damaged,and the TNF-α concentration and TNF-R1 protein expression level were significantly increased(P<0.01). Compared with the OGD/R group, β-asarone(20, 40, 80μg/mL) all showed an increase in cell survival rate and a decrease in LDH release rate(P<0.01). The difference was that β-asarone 20μg/mL had the best effect. β-asarone 20μg/mL can improve cell morphology,and decrease the intracellular TNF-α concentration and TNF-R1 protein expression level(P<0.05). Conclusion: β-asarone can improve the injury of SH-SY5 Y cells induced by OGD/R, and the mechanism may be through the inhibition of TNF-α and TNF-R1 in OGD/R injury to improve neural injury.

关 键 词：Β-细辛醚 SH-SY5Y细胞 氧糖剥夺/复糖复氧 肿瘤坏死因子-Α 肿瘤坏死因子受体-1 

分 类 号：R285[医药卫生—中药学]