α-细辛醚、β-细辛醚改善Aβ_(25-35)诱导的PC12细胞损伤及机制  

Protective effect ofα-asarone andβ-asarone on Aβ_(25-35)-induced inflammatory response in PC12 cells and its mechanism

作　　者：石坚宏 李瑞芝[2] 杨元宵 姬丽婷 李昌煜[1] SHI Jianhong;LI Ruizhi;YANG Yuanxiao;JI Liting;LI Changyu(College of Pharmaceutical Science,Zhejiang Chinese Medical University,Hangzhou 310053,China;Department cf Science and Education,Haining Traditional Chinese Medicine Hospital,Jiaxing 314400,Zhejiang Province,China;School of Pharmacy,Hangzhou Medical College,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学药学院,浙江杭州310053 [2]海宁市中医院科教科,浙江嘉兴314400 [3]杭州医学院药学院,浙江杭州310053

出　　处：《浙江大学学报（医学版）》2021年第5期591-600,共10页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省自然科学基金(Q21H280028)。

摘　　要：目的:探究α-细辛醚、β-细辛醚对β淀粉样蛋白活性片段Aβ_(25-35)诱导的PC12细胞损伤模型的保护作用及相关作用机制。方法:采用Aβ_(25-35)诱导PC12细胞建立Aβ毒性损伤细胞模型。将PC12细胞分为空白对照组、模型对照组、α-细辛醚组(0.5、1.0、1.5μg/mL)、β-细辛醚组(6.3、12.5、25.0μg/mL)、血管活性肠肽(VIP)组,并设VIP拮抗剂对照。采用细胞计数试剂盒(CCK-8)法检测细胞存活率;流式细胞术检测细胞凋亡率;酶联免疫吸附试验检测炎症因子白介素(IL)-1、IL-10、肿瘤坏死因子(TNF)-α,氧化因子诱生型一氧化氮合酶(iNOS)、一氧化氮(NO)和凋亡因子caspase-3、p53水平;蛋白质印迹法检测细胞c-Jun氨基端激酶(JNK)、p38丝裂原活化蛋白激酶(p38MAPK)蛋白表达。结果:与模型对照组比较,α-细辛醚组、β-细辛醚组和VIP组细胞存活率增加,细胞凋亡率下降,凋亡因子caspase-3、p53和炎症因子IL-1、TNF-α水平下降,IL-10水平升高,氧化因子iNOS和NO水平下降,c-Jun氨基端激酶(JNK)、p38MAPK蛋白表达减少(均P<0.05)。VIP拮抗剂干预后,β-细辛醚组细胞存活率下降,细胞凋亡率增加,凋亡因子caspase-3、p53和炎症因子IL-1、TNF-α水平升高,IL-10水平下降,氧化因子i NOS和NO水平升高,JNK、p38MAPK蛋白表达增加(均P<0.05);α-细辛醚组无显著变化(均P>0.05)。结论:α-细辛醚、β-细辛醚对Aβ_(25-35)诱导的PC12细胞损伤模型具有保护作用,β-细辛醚可通过促进VIP的分泌,调控JNK/MAPK通路从而抑制炎症因子、氧化因子水平,改善PC12细胞凋亡;α-细辛醚作用机制与VIP分泌水平无明显联系。Objective: To investigate effects of α-asarone and β-asarone on Aβ_(25-35)-induced PC12 cell injury and related mechanisms.Methods: Aβ toxic injury cell model was induced byAβ_(25-35) in PC12 cells. PC12 cells were divided into blank control group,model control group,α-asarone group(0.5, 1.0, 1.5μg/mL),β-asarone group(6.3, 12.5,25.0μg/mL), vasoactive intestinal peptide(VIP) group, and VIP antagonist control group.Cell survival rate was detected by CCK-8 kit;cell apoptosis rate was detected by flow cytometry. The levels of inflammatory cytokines interleukin(IL)-1, IL-10, tumor necrosis factor(TNF)-α, oxidation-related inducible nitric oxide synthase(iNOS), nitric oxide(NO),apoptosis factors caspase-3 and p53 were detected by ELISA method. The expressions of C-Jun N-terminal kinase(JNK) and p38 mitogen-activated protein kinase(p38 MAPK)were detected by Western blotting.Results: Compared with model control group, cell survival rates ofα-asarone group,β-asarone group and VIP group increased;the cell apoptosis rate decreased;levels of apoptosis-related factors caspase-3, p53, inflammatory factors IL-1, TNF-αdecreased;IL-10 level increased;levels of oxidization-related factors iNOS and NO decreased;the expression of JNK and p38 MAPK protein decreased(all P<0.05). After VIP antagonist intervention, the survival rate ofβ-asarone group decreased;apoptosis rate increased;apoptosis related factors caspase-3, p53, inflammatory factors IL-1, TNF-αincreased;IL-10 decreased;oxidation related factors iNOS and NO increased;the expression of JNK and p38 MAPK protein increased(allP<0.05);while there were no significant changes in these indicators ofα-asarone group(allP>0.05).Conclusion:α-asarone andβ-asarone have protective effects on PC12 cell injury induced byAβ_(25-35) β-asarone may inhibit inflammatory factors and oxidation-related factors through promoting VIP secretion, regulating JNK/MAPK pathway, and reducing PC12 cell apoptosis;however, the effect ofα-asarone may be not related to VIP secretion.

关 键 词：阿尔茨海默病 Α-细辛醚 Β-细辛醚 PC12细胞 炎症 氧化因子 JNK/MAPK通路 血管活性肠肽 

分 类 号：R285.5[医药卫生—中药学]