冬凌草甲素调节IL-6/STAT3信号通路对氧糖剥夺/复氧小胶质细胞增殖、自噬、凋亡及炎症反应的影响OA
The effect of oridonin regulating the IL-6/STAT3 signaling pathway on proliferation,autophagy,apoptosis,and inflammatory response in microglia subjected to oxygen-glucose deprivation/reoxygenation
[目的]探讨冬凌草甲素(ORI)调节白细胞介素(IL)-6/转录活化因子3(STAT3)信号通路对氧糖剥夺/复氧(OGD/R)小胶质细胞增殖、自噬、凋亡及炎症反应的影响.[方法]将小胶质细胞BV2细胞分成:对照组、OGD/R组、ORI低、中、高浓度(ORI-L、ORI-M、ORI-H)组、ORI与重组IL-6联合处理组.各组给予相应干预并培养24 h.细胞计数试剂盒8(CCK8)法和5-乙炔基-2'-脱氧尿苷(EdU)染色法检测BV2细胞增殖;透射电子显微镜观察BV2细胞自噬;流式细胞仪检测BV2细胞凋亡;酶联免疫吸附法检测BV2细胞中炎症因子[肿瘤坏死因子-α(TNF-α)、IL-18、IL-1β]水平;蛋白印迹法检测BV2细胞中IL-6、STAT3蛋白表达.[结果]与对照组比较,OGD/R组BV2细胞存活率、EdU阳性细胞率降低,自噬空泡数、凋亡率、TNF-α、IL-18、IL-1β、IL-6、STAT3蛋白表达升高(P<0.01);与OGD/R组比较,ORI-L组、ORI-M组、ORI-H组BV2细胞存活率、EdU阳性细胞率呈浓度依赖性升高,自噬空泡数、凋亡率、TNF-α、IL-18、IL-1β、IL-6、STAT3蛋白表达呈浓度依赖性降低(P<0.05);与ORI-H组比较,ORI与重组IL-6联合处理组BV2细胞存活率、EdU阳性细胞率降低,自噬空泡数、凋亡率、TNF-α、IL-18、IL-1β、IL-6、STAT3蛋白表达升高(P<0.01).[结论]ORI能够促进OGD/R诱导的小胶质细胞增殖,抑制OGD/R诱导的小胶质细胞自噬、凋亡及炎症反应,其机制可能是通过抑制IL-6/STAT3信号通路实现的.
[Objective]To investigate the effect of oridonin(ORI)on proliferation,autophagy,apoptosis,and inflammatory response in microglia subjected to oxygen-glucose deprivation/reoxygenation(OGD/R)and its potential mechanism involving the interleukin-6(IL-6)/signal transducer and activator of transcription 3(STAT3)signaling pathway.[Methods]BV2 microglial cells were divided into the following groups:control,OGD/R,ORI(low,medium,and high concentration:ORI-L,ORI-M,ORI-H),and ORI combined with recombinant IL-6.After respective treatments for 24 h,cell proliferation was assessed by Cell Counting Kit-8(CCK-8)assay and 5-ethynyl-2′-deoxyuridine(EdU)staining.Autophagy was observed under a transmission electron microscope.Apoptosis was detected by flow cytometry.The levels of inflammatory factors[tumor necrosis factor-α(TNF-α),IL-18,and IL-1β]were measured using enzyme-linked immunosorbent assay(ELISA).The protein expression levels of IL-6 and STAT3 were determined by Western blotting.[Results]Compared with the control group,the OGD/R group showed decreased cell viability and EdU-positive cell rate,along with increased number of autophagic vacuoles,apoptosis rate,levels of TNF-α,IL-18,IL-1β,and protein expression of IL-6 and STAT3(P<0.01).Compared with the OGD/R group,the ORI-L,ORI-M,and ORI-H groups exhibited a concentration-dependent increase in cell viability and EdU-positive cell rate,and a concentration-dependent decrease in the number of autophagic vacuoles,apoptosis rate,levels of TNF-α,IL-18,IL-1β,and protein expression of IL-6 and STAT3(P<0.05).Compared with the ORI-H group,the ORI combined with recombinant IL-6 group demonstrated decreased cell viability and EdU-positive cell rate,and increased number of autophagic vacuoles,apoptosis rate,levels of TNF-α,IL-18,IL-1β,and protein expression of IL-6 and STAT3(P<0.01).[Conclusion]ORI can promote proliferation and inhibit autophagy,apoptosis,and inflammatory response in OGD/R-induced microglia.Its mechanism may be associated with the inhibition of the IL-6/STAT3 signaling pathway.
李林;李向男;杨松涛;李建民;张云鹤;付爱军;张志勇
华北理工大学附属医院神经外科,唐山 063015华北理工大学附属医院神经外科,唐山 063015华北理工大学附属医院神经外科,唐山 063015华北理工大学附属医院神经外科,唐山 063015华北理工大学附属医院神经外科,唐山 063015华北理工大学附属医院神经外科,唐山 063015华北理工大学附属医院神经外科,唐山 063015
医药卫生
冬凌草甲素白细胞介素-6/转录活化因子3信号通路氧糖剥夺/复氧小胶质细胞增殖自噬凋亡
oridoninIL-6/STAT3 signaling pathwayoxygen-glucose deprivation/reoxygenationmicrogliaproliferationautophagyapoptosis
《天津中医药大学学报》 2026 (3)
301-307,7
河北省医学科学研究课题计划项目(20240045).
评论