辣椒素对氧葡萄糖剥夺/再灌注所致SH-SY5Y细胞损伤的保护作用及机制研究  

Protective Effect and Mechanism of CAP on SH-SY5Y Cell Injury Induced by OGD/R

作　　者：马世江[1] 沈长波[1] 刘杰[1] 谭军[1] MA Shijiang;SHEN Changbo;LIU Jie;TAN Jun(The Third Affiliated Hospital of Xinxiang Medical University,Xingxiang 453003,China)

机构地区：[1]新乡医学院第三附属医院,河南新乡453003

出　　处：《中医药信息》2021年第9期40-47,共8页Information on Traditional Chinese Medicine

基　　金：新乡市科技局科技创新人才计划项目(CXRC17003)。

摘　　要：目的:研究辣椒素(CAP)对氧葡萄糖剥夺/再灌注(Oxygen-glucosedeprivation/reperfusion,OGD/R)所致SH-SY5Y细胞损伤的保护作用及可能机制。方法:将SH-SY5Y细胞随机分为6组:正常组,模型组,尼莫地平组(5μg/mL)和CAP低、中、高剂量组(1、5、25μmol/L)。正常组细胞不进行OGD/R干预,一直在正常条件下培养,其余各组细胞通过OGD/R建立细胞损伤模型。在整个OGD/R过程中,分别给予尼莫地平组和CAP低、中、高剂量组相应剂量的药物。OGD/R 24 h后,分别检测各组细胞的细胞活力、乳酸脱氢酶(LDH)水平、Caspase-3和Caspase-9活性、Bcl-2和BaxmRNA的表达,同时采用网络药理学方法探讨CAP治疗缺血性卒中的潜在作用机制,并进行实验验证。结果:与正常组相比,OGD/R导致SH-SY5Y细胞活力降低,Bcl-2 mRNA表达降低,LDH、Caspase-3和Caspase-9活性升高,BaxmRNA的表达增高(P<0.01)。CAP则能明显增强OGD/R作用下SH-SY5Y细胞活力,降低LDH、Caspase-3和Caspase-9活性,提高Bcl-2 mRNA表达,降低BaxmRNA的表达(P<0.01)。通过网络药理学方法共获得CAP抗缺血性卒中的相关基因59个,经KEGG富集分析发现,PI3K/Akt信号通路富集最显著。实验验证发现,抑制PI3K可显著降低CAP的保护作用,同时与正常组相比,CAP中、高剂量组可显著增加PI3K、AktmRNA表达水平(P<0.05,P<0.01)。结论:CAP能减轻OGD/R诱导的SH-SY5Y细胞损伤,减少细胞凋亡,这可能与激活PI3K信号通路有关。CAP可能是一种有前途的治疗缺血性卒中的化合物。Objective:To study the protective effect of Capsaicin(CAP)on SH-SY5Y cells injured by oxygen-glucose deprivation/reperfusion(OGD/R)and its possible mechanism.Methods:SH-SY5Y cells were randomly divided into the normal group,the model group,the Nimodipine group(5μg/mL)and the CAP groups of low-dose,medium-dose and high-dose(1,5,25μmol/L).The cells in the normal group were not exposed to OGD/R and were cultured under normal conditions.The cells in the other groups were established cell injury model by OGD/R.Nimodipine and CAP were administered throughout the OGD/R process.After 24 hours of OGD/R intervention,the viability of cells,the level of lactate dehydrogenase(LDH),the activities of Caspase-3 and Caspase-9,as well as the mRNA expressions of Bcl-2 and Bax were detected in the groups.Network pharmacology method was used to explore the potential mechanism of CAP in the treatment of ischemic stroke.Results:OGD/R decreased the activities of SH-SY5Y cells,reduced Bcl-2 mRNA expression,enhanced the activities of LDH,Caspase-3 and Caspase-9,and increased Bax mRNA expression(P<0.01).CAP could significantly enhance the activity of SH-SY5Y cells,reduce the activities of LDH,Caspase-3 and Caspase-9,increase the expression of Bcl-2 mRNA and decrease the expression of Bax mRNA under the action of OGD/R(P<0.01).A total of 59 genes related to CAP against ischemic stroke were obtained by network pharmacology.KEGG enrichment analysis showed that PI3K/Akt signaling pathway was the most significant one.The experimental results showed that inhibition of PI3K could significantly reduce the protective effect of CAP,and the expression levels of PI3K and Akt mRNA were significantly increased in the CAP groups of medium-dose and high-dose compared to those in the normal group(P<0.05,P<0.01).Conclusion:CAP can reduce the damage of SY5Y cells induced by OGD/R and reduce apoptosis,which may be related to activating PI3K signaling pathway.CAP may be a promising therapeutic agent for ischemic stroke.

关 键 词：辣椒素 氧葡萄糖剥夺/再灌注 细胞凋亡 PI3K 

分 类 号：R285.5[医药卫生—中药学]