仙茅苷调节JAK2/STAT3信号通路对红藻氨酸体外诱导的小胶质细胞损伤的影响  

Effect of Curculigoside on Kainic Acid Induced Microglia Injury by Regulating JAK2/STAT3 Signaling Pathway In Vitro

作　　者：尚杨 张墨 张俊娜[1] 侯国盛[2] 阮晓兰[2] SHANG Yang;ZHANG Mo;ZHANG Junna;HOU Guosheng;RUAN Xiaolan(Department of Emergency,Linxi Hospital,Kailuan General Hospital,Tangshan 063000,China;Department of Neurology,Linxi Hospital,Kailuan General Hospital,Tangshan 063000,China)

机构地区：[1]开滦总医院林西医院急诊科,唐山063000 [2]开滦总医院林西医院神经内科,唐山063000

出　　处：《中国细胞生物学学报》2025年第2期203-211,共9页Chinese Journal of Cell Biology

基　　金：河北省2024年度医学科学研究课题计划(批准号:20241273)资助的课题。

摘　　要：该文探究仙茅苷(CCG)调节Janus蛋白酪氨酸激酶2(JAK2)/信号转导和转录激活因子3(STAT3)信号通路对红藻氨酸(KA)体外诱导的小胶质细胞损伤的影响。将HMC3细胞随机分为HMC3组、KA组、L-CCG组、M-CCG组、H-CCG组、H-CCG+JAK2/STAT3通路激活剂colivelin组,HMC3组不做干预,KA组用600μmol/L KA刺激,L-CCG组、M-CCG组、H-CCG组在KA组基础上分别加5、10、20μmol/L CCG,H-CCG+colivelin组在H-CCG组基础上加0.5μmol/L colivelin。Ed U法检测细胞增殖情况;Hochest 33258/PI法检测细胞凋亡情况;DCFH-DA法检测活性氧(ROS)表达情况;γ组蛋白H2A变异体(γ-H2AX)免疫荧光法检测DNA损伤情况;试剂盒检测细胞中烟酰胺腺嘌呤二核苷磷酸/还原型烟酰胺腺嘌呤二核苷酸磷酸(NADP+/NADPH)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、IL-6水平;免疫印迹法检测磷酸化JAK2(p-JAK2)、磷酸化STAT3(p-STAT3)、NOD样受体热蛋白结构域相关蛋白3(NLRP3)、含半胱氨酸的天冬氨酸蛋白水解酶1(Caspase-1)、凋亡相关斑点样蛋白(ASC)表达情况。结果显示,与HMC3组比,KA组HMC3细胞增殖率降低,凋亡率、γ-H2AX核焦点数量、NADP+/NADPH、ROS、IL-1β、TNF-α、IL-6水平增加(P<0.05);与KA组比,L-CCG组、M-CCG组、H-CCG组增殖率依次增加,凋亡率、γ-H2AX核焦点数量、NADP+/NADPH、ROS、IL-1β、TNF-α、IL-6水平依次降低(P<0.05);与H-CCG组比,H-CCG+colivelin组增殖率降低,凋亡率、γ-H2AX核焦点数量、NADP+/NADPH、ROS、IL-1β、TNF-α、IL-6水平增加(P<0.05)。与HMC3组比,KA组HMC3细胞中p-JAK2、p-STAT3、NLRP3、Caspase-1、ASC表达水平增加(P<0.05);与KA组比,L-CCG组、M-CCG组、H-CCG组p-JAK2、p-STAT3、NLRP3、Caspase-1、ASC表达水平依次降低(P<0.05);与H-CCG组比,H-CCG+colivelin组p-JAK2、p-STAT3、NLRP3、Caspase-1、ASC表达水平增加(P<0.05)。总之,CCG可能通过抑制JAK2/STAT3通路对KA诱导的小胶质细胞功能损伤发挥保护作用。The aim of this study was to investigate the effect of CCG(curculigoside)on microglia injury induced by KA(kainic acid)in vitro through regulating the JAK2(Janus kinase-2)/STAT3(signal transducer and activator of transcription 3)signaling pathway.HMC3 cells were randomly divided into HMC3 group,KA group,L-CCG group,M-CCG group,H-CCG group,and H-CCG+colivelin(JAK2/STAT3 pathway activator)group.The cells in the HMC3 group were not intervened,the cells in the KA group were stimulated with 600μmol/L KA,the cells in the L-CCG,M-CCG,and H-CCG groups were added with 5,10,and 20μmol/L CCG,respectively,and the cells in the H-CCG+colivelin group were added with 0.5μmol/L colivelin on the basis of the H-CCG group.The cell proliferation was detected by EdU method.Apoptosis was detected by Hochest 33258/PI method.The expression of ROS(reactive oxygen species)was detected by DCFH-DA method.DNA damage was detected byγ-H2AX(γhistone family 2A variant)immunofluorescence method.The levels of NADP+/NADPH(nicotinamide adenine dinucleoside phosphate/reduced nicotinamide adenine dinucleotide phosphate),IL-1β(interleukin-1β),TNF-α(tumor necrosis factor-α),and IL-6 in cells were detected by kit.The expression of p-JAK2(phosphorylated JAK2),p-STAT3(phosphorylated STAT3),NLRP3(NOD-like receptor thermal protein domain associated protein 3),Caspase-1(cysteinyl aspartate specific proteinase 1),and ASC(apoptosis-associated speck-like protein)were detected by Western blot.The results showed that compared with the HMC3 group,the proliferation rate of HMC3 cells in the KA group was decreased,while the apoptosis rate,number ofγ-H2AX nuclear foci,and the levels of NADP+/NADPH,ROS,IL-1β,TNF-α,and IL-6 were increased(P<0.05).Compared with the KA group,the proliferation rate of the LCCG group,M-CCG group,and H-CCG group were increased successively,while the apoptosis rate,number ofγ-H2AX nuclear foci,and the levels of NADP+/NADPH,ROS,IL-1β,TNF-α,and IL-6 were decreased successively(P<0.05).Compared with the H-CCG group,the proliferatio

关 键 词：仙茅苷 Janus蛋白酪氨酸激酶2/信号转导和转录激活因子3通路 红藻氨酸 小胶质细胞功能损伤 

分 类 号：R285[医药卫生—中药学]