摘要
目的:探讨克罗米芬(CC)对子宫内膜基质细胞(hEndoSCs)的损伤作用,并研究亚精胺对受损子宫内膜基质细胞自噬和炎症细胞因子表达的影响。方法:实验分为对照组、亚精胺组、克罗米芬组(CC组)、CC+亚精胺组。MTT法检测hEndoSCs与不同浓度CC或亚精胺共同孵育24 h后细胞的存活率;流式细胞技术检测4组细胞内活性氧(ROS)含量和细胞凋亡水平。Western blot检测自噬途径相关蛋白ULK1、p-ULK1、LC-3Ⅱ和凋亡相关蛋白Bax、Bcl-2、Cleaved-caspase 3的表达。采用RT-qPCR检测IL-6、IL-1β、TNF-αmRNA表达量。结果:与对照组相比,CC组细胞存活率下降,细胞凋亡率、ROS含量、Bax、Cleaved-caspase 3表达量升高,Bcl-2表达量降低,自噬相关蛋白p-ULK1、LC-3Ⅱ/Ⅰ水平降低,炎症因子IL-6、IL-1β、TNF-αmRNA表达升高(P<0.01)。与对照组相比,亚精胺组细胞的存活率无明显变化(P>0.05)。与CC组相比,CC+亚精胺组细胞存活率显著提高,细胞凋亡率、ROS含量、Bax和Cleaved-caspase 3表达量降低,Bcl-2表达量升高,自噬相关蛋白p-ULK1、LC-3Ⅱ/Ⅰ表达升高,炎症因子IL-6、IL-1β、TNF-αmRNA表达降低(P<0.01)。结论:CC能够抑制子宫内膜基质细胞增殖,促进细胞凋亡,提高炎症因子IL-6、IL-1β、TNF-α的转录水平。亚精胺可通过激活细胞自噬,降低CC损伤的子宫内膜基质细胞胞内ROS水平,提高细胞存活率,并抑制炎症因子IL-6、IL-1β、TNF-α表达。
Objective:To investigate damaging effects of clomifene citrate(CC)on endometrial stromal cells(hEndoSCs),and to study effects of spermidine on autophagy and inflammatory cytokine expression in damaged endometrial stromal cells.Methods:Groups were firstly divided into control group,spermidine group,clomiphene group(CC group),CC+Spermidine group.MTT assay was used to detect cell survival rate of hEndoSCs after co-incubation with different concentrations of CC or Spermidine for 24 h.Content of intracellular reactive oxygen species(ROS)and level of apoptosis in cells of the 4 groups were detected by flow cytometry technique.Western blot was used to detect expressions of autophagy pathway-related proteins ULK1,p-ULK1,LC-3Ⅱ,and apoptosis-related proteins Bax,Bcl-2,Cleaved-caspase 3.RT-qPCR was used to detect mRNA expressions of IL-6,IL-1βand TNF-α.Results:Compared with control group,CC group showed decreased cell survival,increased apoptosis rate,ROS content,Bax,Cleaved-caspase 3 expressions,decreased Bcl-2 expression,decreased levels of autophagy-related proteins p-ULK1 and LC-3Ⅱ/Ⅰ,and elevated expressions of inflammatory factors IL-6,IL-1βand TNF-αmRNA(P<0.01).There was no significant changes viability of cells in spermidine group compared with control group(P>0.05).Compared with CC group,cell survival rate in CC+spermidine group was significantly increased,apoptosis rate,ROS content,Bax and Cleaved-caspase 3 expressions were decreased,Bcl-2 expression was increased,expressions of autophagy-related proteins p-ULK1 and LC-3Ⅱ/Ⅰwere elevated,while expressions of inflammatory factors IL-6,IL-1βand TNF-αmRNA were decreased(P<0.01).Conclusion:CC can inhibit endometrial stromal cell proliferation,promote apoptosis,and increase the transcript levels of inflammatory factors IL-6,IL-1βand TNF-α.Spermidine can reduce intracellular ROS in clomiphene-injured endometrial stromal cells by activating cellular autophagy,increase cell survival,and inhibit the expressions of inflammatory factors IL-6,IL-1βand TNF-α.
作者
张雪
米旭光
林秀英
付建华
刘磊
高心月
方艳秋
ZHANG Xue;MI Xuguang;LIN Xiuying;FU Jianhua;LIU Lei;GAO Xinyue;FANG Yanqiu(Changchun University of Chinese Medicine,Changchun 130021,China;Jilin Provincial People's Hospital Reproductive Medicine Center,Changchun 130021,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2024年第5期1023-1029,共7页
Chinese Journal of Immunology
基金
吉林省科技发展计划项目(20230204036YY,20230203051SF,YDZJ202102CXJD076)
吉林省卫生健康科技能力提升项目(2021lc059)。
关键词
亚精胺
子宫内膜基质细胞
炎症
自噬
细胞凋亡
克罗米芬
Spermine
Endometrial stromal cells
Inflammation
Autophagy
Apoptosis
Clomiphene citrate
